CN103908571B - A kind of Chinese traditional compound medicine for treating heart disease - Google Patents
A kind of Chinese traditional compound medicine for treating heart disease Download PDFInfo
- Publication number
- CN103908571B CN103908571B CN201310006120.8A CN201310006120A CN103908571B CN 103908571 B CN103908571 B CN 103908571B CN 201310006120 A CN201310006120 A CN 201310006120A CN 103908571 B CN103908571 B CN 103908571B
- Authority
- CN
- China
- Prior art keywords
- group
- rhizoma polygonati
- onion bulb
- longstamen onion
- extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
In order to meet clinical needs, preferably treatment cardiovascular and cerebrovascular disease, improve the general level of the health of the people, the present invention provides a kind of new pharmaceutical compositions and preparation method thereof for being used to treat cardiovascular and cerebrovascular disease, the pharmaceutical composition is mainly prepared by leaves of Hawthorn, Longstamen Onion Bulb, rhizoma polygonati, in terms of being used to prepare treatment cardiovascular and cerebrovascular disease, unexpected effect is produced.
Description
Technical field
The invention belongs to pharmaceutical technology field, be related to it is a kind of for treat cardiovascular and cerebrovascular disease mainly by leaves of Hawthorn, Chinese onion
White, preparation of pharmaceutical composition made of rhizoma polygonati and preparation method thereof.
Background technique
Cardiovascular disease, also known as circulation system disease are a series of to be related to the disease of the circulatory system.The circulatory system refers to people
Internal hemophoric organ and tissue, mainly include heart, blood vessel (artery, vein, capilary), can be subdivided into it is acute and
It is chronic, it is typically related with artery sclerosis.
Cardiovascular disease is that one kind seriously threatens the mankind, the common disease of especially 50 years old or more middle-aged and the old's health, with
The change of improvement of living standard and rhythm of life, referred to as " three high diseases " (i.e. hypertension, hyperglycemia and the high blood of " rich people's disease "
Rouge) it is increasing.With advancing age, Prevalence of Hypertension gradually increases.40%~45% suffers from 60 years old or more the elderlys
Hyperglycemia or hyperlipidemia are also suffered from while having hypertension, show that 50% or so diabetes patient merges according to external data
There are a variety of infirmitiess of age such as hypertension, hyperlipidemia.The common pathologic basis such as angina pectoris, myocardial infarction, ischemic heart disease is all
It is myocardial ischemia, myocardial blood supply leads to myocardial metabolism disorder for hypoxgia, and energy supply is insufficient, myocardium shrinkage function decline, blood
Liquid output quantity reduces, and then influences the function of entire body, or even cause cardiomyocyte cell death.Cerebral ischemia re-pouring is clinically
The key of cerebrovascular disease therapy.Energetic supersession, secondary lactic acid accumulation, the exceeded, radical damage of calcium etc. are influenced after cardiac-cerebral ischemia
A variety of variations；Multiple target point reverses or improves these variations, improves the important goal that comprehensive therapeutic effect is drug therapy.
Leaves of Hawthorn be rosaceous plant large-fruited Chinese hawthorn Crataegus pinnatifida Bge.var.major N.E.Br. or
The dried leaf of hawthorn Crataegus pinnatifida Bge..Leaves of Hawthorn main component is vetexin-glucoside, Vitexin
A variety of chemical components such as rhamnoside, rutin, Vitexin, Hyperoside, ursolic acid, Quercetin and Vitexin, pharmacological action master
Be embodied in anti-inflammatory and antalgic, reducing blood lipid, prevention and treatment atherosclerosis, the protection to myocardial ischemia, the protection to cerebral ischemia, to liver
It is dirty to protect, to protection renal, inhibition tumour cell etc..
Longstamen Onion Bulb is liliaceous plant Allium macrostemon Allium macrostemon Bge. or Chinese onion Allium chinensis
The dry bulb of G.Don.Longstamen Onion Bulb main component is the fatty acid composition, total of the compositions such as linoleic acid, palmitinic acid, oleic acid and sitosterol
Sapogenin and total alkaloid.The pharmacological action of Longstamen Onion Bulb mainly has inhibiting bacteria and diminishing inflammation, spasmolysis to relieving asthma, platelet aggregation-against, anti-oxidant, drop
Blood lipid, antiatherosclerosis, it is antitumor the effects of.
Rhizoma polygonati is liliaceous plant polygonatum kingianurn Polygonatum kingianum Coll.et Hemsl., rhizoma polygonati
The dry rhizome of Polygonatum sibiricum Red. or polygonatum cyrtonema Polygonatum cyrtonema Hua.Rhizoma polygonati
Main component is polysaccharide, steroid saponin, flavones, anthraquinone analog compound, amino acid isoreactivity ingredient.The pharmacological action of rhizoma polygonati is main
There is hypoglycemic, the protection to myocardial damage, the protection to cerebral ischemia, anti-oxidant, anti-aging improves memory, anti-swollen
Tumor, it is antifatigue, it is antibacterial and anti-oxidant etc..
Currently with leaves of Hawthorn, the interaction of Longstamen Onion Bulb, rhizoma polygonati, composition of prescription is for treating cardiovascular and cerebrovascular disease, also not
It appears in the newspapers.
Summary of the invention
In order to meet clinical needs, cardiovascular and cerebrovascular disease is preferably treated, improves the general level of the health of the people, the present invention provides
A kind of new for treating pharmaceutical composition of cardiovascular and cerebrovascular disease and preparation method thereof, the pharmaceutical composition is mainly by hawthorn
Leaf, Longstamen Onion Bulb, rhizoma polygonati are prepared, and in terms of being used to prepare treatment cardiovascular and cerebrovascular disease, produce unexpected effect.
Pharmaceutical composition of the present invention is mainly prepared by leaves of Hawthorn, Longstamen Onion Bulb, rhizoma polygonati, the parts by weight of bulk pharmaceutical chemicals are as follows:
1~100 part of leaves of Hawthorn, 1~100 part of Longstamen Onion Bulb, 1~100 part of rhizoma polygonati；It is preferred that are as follows: 1~10 part of leaves of Hawthorn, 1~4 part of Longstamen Onion Bulb, rhizoma polygonati
1~2 part；It is optimal are as follows: 7 parts of leaves of Hawthorn, 3.5 parts of Longstamen Onion Bulb, 1.5 parts of rhizoma polygonati.
Leaves of Hawthorn, Longstamen Onion Bulb, rhizoma polygonati in pharmaceutical composition described above can be mentioned with suitable solvent and method list or
Extract is prepared in mixed mention, and total extract is mixed and made into any one preparation with pharmaceutically acceptable auxiliary material again.It is wherein described
The preferred water of Extraction solvent or ethyl alcohol, extracting method can for infusion process, percolation, decocting method, reflux extraction or continuously mention
It follows the example of.
The present invention provides leaves of Hawthorn Optimized extraction techniques, hawthorne leaf P.E can be made according to by following methods, but
It is not limited only to following methods:
7 parts of leaves of Hawthorn are weighed, the solvent soaking time 0.1~10 hour for using 2 times~40 times of medicinal material total weight for the first time
Afterwards, it extracts 0.1~10 hour under conditions of 30 DEG C~100 DEG C, is existed for the second time with 2 times~20 times of solvent of medicinal material total weight
30 DEG C~100 DEG C of condition extracts 0.1~10 hour, for the third time with 2 times~20 times amount solvents of medicinal material total weight 30 DEG C~
100 DEG C of condition is extracted 0.1~10 hour, and the 4th 2 times~20 times amount solvents with medicinal material total weight are at 30 DEG C~100 DEG C
Condition is extracted 0.1~10 hour, combined extract, is distilled to recover solvent, distillate centrifugation, with 0.1 times of medicinal material total weight~
Three times, the supernatant of supernatant and washing precipitating after merging centrifugation, distillation and concentration is to close for the solvent washing precipitating of 100 times of volumes
Dry, pulverize after dry to get.
It is 10%~35% by extract yield prepared by above-mentioned technique, the content of general flavone is with anhydrous rutin
(C21H20O12) meter, it is not less than 20%；Vitexin rhamnoside (C27H34O14) content be not less than 2%.
The present invention provides Longstamen Onion Bulb, the Optimized extraction techniques of rhizoma polygonati, Longstamen Onion Bulb, Rhizoma Polygonati extract can be according to by following methods
It is made, but is not limited only to following methods:
3.5 parts and 1.5 parts of rhizoma polygonati of Longstamen Onion Bulb are weighed, 2 times of medicinal material total weight are added for the first time under the conditions of 50 DEG C~100 DEG C
~40 times solvent extraction 0.1~10 hour, second be added medicinal material total weight 2 times~20 times solvent extraction 0.1~10 hour,
Collecting decoction, filtrate are concentrated into 0.1 times~20 times of medicinal material total weight, and ethyl alcohol is added to make alcohol content 20%~90%, it is static or from
The heart takes supernatant, filtering, be distilled to recover ethyl alcohol, continue distillation and concentration at thick paste, dry, pulverize to get.
The Longstamen Onion Bulb that is prepared by above-mentioned technique, Rhizoma Polygonati extract yield are 25%~45%, containing polysaccharide with DEXTROSE ANHYDROUS
(C6H12O6) count not less than 4%.
Composition described above can increase or reduce according to corresponding proportion in production by weight as matching, such as big rule
Mould production can with kilogram for raw material, or as unit of ton, small-scale production can also in grams, weight can increase or
Person reduce, but between each component weight proportion constant rate.
The ratio of the above weight proportion is obtained by science screening, for especial patient, can accordingly adjust composition
Ratio, increase or reduce be no more than 100%.
The dosage of medicine components of the present invention be carried out by inventor a large number of experiments grope summarize obtain, each component dosage
All there is good therapeutic effect within the scope of above-mentioned parts by weight.
The present invention provides a kind of pharmaceutical compositions being used to prepare in terms for the treatment of cardiovascular and cerebrovascular disease, are mainly used for treating
Cerebral thrombosis, coronary heart diseases and angina pectoris, vasculitis, myocardial infarction and hyperlipidemia etc. disease.
Pharmaceutical composition of the present invention can add one or more pharmaceutically acceptable carriers, with oral and extra-parenteral administration
Mode be applicable to the patient of this treatment.When for taking orally, it can be made into conventional solid pharmaceutical preparation, such as tablet, glue
Capsule, dispersible tablet, oral solution, particle, chewable tablets, oral disintegrating tablet, dripping pill, sustained release tablets, spansule, controlled release tablet, controlled release capsule, are made
Liquid preparation such as water or oil-suspending agent or other liquid preparations such as syrup etc.；When for parenteral administration, injection can be made into
Solution, water or oil-suspending agent etc., such as water needle, freeze-dried powder, aseptic powder injection, infusion.Preferred form is injection, piece
Agent and capsule.
The production of the conventional method in existing pharmaceutical field can be used in pharmaceutical composition of the present invention, can add when needs
Various pharmaceutically acceptable carriers.The carrier includes the diluent, excipient, filler, bonding of pharmaceutical field routine
Agent, wetting agent, disintegrating agent, sorbefacient, surfactant, absorption carrier, lubricant etc..
Pharmaceutical composition of the present invention is when being made injection, and in order to increase its solubility, the solubilisings such as Tween-80 can be added
Agent.The isotonic regulator for adjusting osmotic pressure can be added in infusion, for example, sodium chloride, potassium chloride, magnesium chloride, calcium chloride,
Sodium lactate, glucose, xylitol, sorbierite and dextran etc., preferably sodium chloride or glucose.Figuration can be added in powder needle
Agent, for example, mannitol, glucose etc..
Pharmaceutical composition of the present invention has the advantage that
(1) it provides a kind of new for treating pharmaceutical composition of cardiovascular and cerebrovascular disease and preparation method thereof, meets
Urgent clinical needs.
(2) interaction to pharmaceutical composition of the present invention for the first time and composition of prescription have carried out pharmacodynamic study, as a result such as
Under:
It is raised 1. the pharmaceutical composition can improve S-T segment；Creatine kinase in Serum fibrosis markers (CK), lactic acid can be substantially reduced
The activity of dehydrogenase (LDH), aspartate amino transferase (AST)；Reduce atpase activity, lactic acid (LD) and free rouge
Fat acid (NEFA) content reduces；Reduce malonaldehyde (MDA) content, total number born (SOD), glutathione peroxidating
The activity of object enzyme (GSH-Px) increases；Each administration group whole blood viscosity reduces, plasma viscosity reduces, hematocrit reduces, erythrocyte sedimentation rate
It reduces, illustrates that the pharmaceutical composition can inhibit erythrocyte aggregation, reduce erythrocyte fragility, enhance its morphotropism, make the raised heart
Rate tends to be normal, left ventricular systolic pressure increases, left room diastolic pressure reduces, reduction of speed under left room maximum climbing speed raising, left room maximum
Rate increases, terminal diastolic pressure reduces.Treatment group's lesion and the obvious mitigation cardiac muscle cell disorder of model group, breaking degree compared with
Gently, karyopycnosis and eosinophilic cytoplasmic become apparent reduction, and myocardial infarction area significantly reduces, and illustrate that the pharmaceutical composition lacks cardiac muscle
The improvement and therapeutic effect that the change of blood rat model indices has.
2. the pharmaceutical composition can significantly reduce total cholesterol in serum (TC), triglycerides (TG), low-density lipoprotein
The content of cholesterol (LDL-C) and apolipoprotein B (ApoB) increases high-density lipoprotein cholesterol (HDL-C) and apolipoprotein
A1 (ApoA1) content；Malonaldehyde (MDA) content can be significantly reduced, increases the content of nitric oxide (NO), increases total superoxides
Mutase (SOD), glutathione peroxidase (GSH-Px) activity；Treatment group's fatty degeneration of liver Leukopenia, endochylema lactones
Drop is reduced or is disappeared, and cell volume organizes liver cell close to normal.Illustrate that the pharmaceutical composition is each to Hyperlipemia model rat
The change of item index has improvement and therapeutic effect.
3. test result, which shows that medicament composition capsule agent curative effect of the present invention is substantially better than, is applied alone leaves of Hawthorn, Longstamen Onion Bulb, rhizoma polygonati.
Prompt leaves of Hawthorn, Longstamen Onion Bulb, rhizoma polygonati compatibility are using having the function of synergy, as a result, the ordinary skill people of the art
Member institute is unexpected.
(3) present composition is respectively matched and has carried out pharmacodynamic study, obtained the optimal proportion of the present composition.
(4) present invention is fed intake with raw material, and preparation process is simple, and quality difference is small between different batches drug, and drug quality is more
It is uniform and stable.
(5) acute toxicity testing carried out shows that the maximal tolerance dose of medicament composition capsule agent of the present invention is equivalent to 70kg
356 times of weight day for human beings research on maximum utilized quantity show pharmaceutical composition low toxicity of the present invention, highly-safe.
(6) stability experiment carried out shows that medicament composition capsule agent indices of the present invention are more stable, guarantees
The safety of clinical application.
(7) present composition combination drug is curative for effect, and reduces opposite dosage, before having a wide range of applications
Scape.
Carry out the beneficial effect of the present invention is further explained described pharmaceutical composition below by way of experimental example.
Embodiment
The specific embodiment of form by the following examples makees further specifically above content of the invention
It is bright.But this should not be interpreted as to the scope of the above subject matter of the present invention is limited to the following embodiments.It is all above-mentioned based on the present invention
The technology that content is realized belongs to the range of invention.The auxiliary material of each dosage form can be with pharmaceutically acceptable auxiliary in above example
Material replacement, or reduce, increase.
Embodiment 1: the preparation of hawthorne leaf P.E
7 parts of leaves of Hawthorn are weighed, for the first time with 8 times of 45% ethyl alcohol soaking time of medicinal material total weight after 2 hours, at 80 DEG C
Under conditions of extract 2 hours, second is extracted 1 hour with 5 times of 45% ethyl alcohol of medicinal material total weight in 80 DEG C of condition, third
It is secondary to extract 0.5 hour in 80 DEG C of condition with 4 times of medicinal material total weight 45% ethyl alcohol of amount, the 4th 3 times with medicinal material total weight
The condition that 45% ethyl alcohol is measured at 80 DEG C is extracted 0.5 hour, and combined extract is distilled to recover ethyl alcohol, and distillate centrifuge is every
It is centrifuged 30min under the revolving speed of minute 3000R, is precipitated three times with the purifying water washing of 2 times of volumes of medicinal material total weight, after merging centrifugation
Supernatant and washing precipitating supernatant, distillation and concentration to dry, pulverize after close dry to get.
The identification of hawthorne leaf P.E
This product 50mg is taken, adds ethyl alcohol 5ml, shakes up, is ultrasonically treated 5 minutes, filtration takes filtrate as test solution.Separately
Control substance of Rutin, Hyperoside reference substance are taken, respectively plus solution of every 1ml containing 0.2mg is made in ethyl alcohol, as reference substance solution.
Tested according to thin-layered chromatography (" Chinese Pharmacopoeia " annex VI B in 2010), draw above-mentioned each 1 μ l of three kinds of solution, put respectively in
On same polyamide film, with ethanol-acetone-water (7: 5: 6) for solvent, it is unfolded, takes out, dry, spray is tried with alchlor
Liquid, drying are set and are inspected under ultraviolet lamp (365nm) after placing 1 hour.In sample chromatogram, with reference substance chromatography corresponding positions
It sets, shows the fluorescence spot of same color.
The assay of hawthorne leaf P.E
The preparation of reference substance solution is accurate to weigh the control substance of Rutin 25mg being dried under reduced pressure at 120 DEG C to constant weight, sets 50ml
In measuring bottle, add appropriate amount of ethanol, ultrasonic treatment makes to dissolve, lets cool, be diluted to scale with ethyl alcohol, shake up, and precision measures 20ml, sets
In 50ml measuring bottle, scale is added water to, is shaken up to get (every 1ml contains anhydrous rutin 0.20mg).
The preparation precision of standard curve measures reference substance solution 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, sets 25ml amount respectively
In bottle, 6ml is respectively added water to, adds 5% sodium nitrite solution 1ml, makes to mix, placed 6 minutes, add 10% aluminum nitrate solution 1ml, shake
It is even, it places 6 minutes, adding sodium hydroxide test solution 10ml is added water to scale, shaken up, and is placed 15 minutes, is with corresponding reagent
Blank is inhaled according to UV-VIS spectrophotometry (" Chinese Pharmacopoeia " annex VA in 2010) with measuring at the wavelength of 500nm
Luminosity is abscissa by ordinate, concentration of absorbance, draws standard curve.
Measuring method takes this product 0.15g, accurately weighed, sets in stuffed conical flask, and Diluted Alcohol 25ml is added in precision, and close plug shakes
It is even, it is ultrasonically treated 5 minutes, place 3 hours or more, filtration, precision measures subsequent filtrate 2ml, sets in 25ml measuring bottle, is diluted with water to
Scale shakes up, as test solution.Precision measures test solution 2ml, sets in 25ml measuring bottle, the preparation of sighting target directrix curve
Method under measures absorbance, while accurate measurement test solution 2ml in accordance with the law, sets 25ml amount from " adding water to 6ml "
In bottle, scale is added water to, is shaken up, as blank solution.From the amount for reading rutin in test solution on standard curve, calculate,
To obtain the final product.
Vitexin rhamnoside
Chromatographic condition and system suitability are using octadecylsilane chemically bonded silica as filler；With tetrahydrofuran-first
Alcohol-acetonitrile-acetic acid-water (38: 3: 3: 4: 152) is mobile phase；Detection wavelength is 330nm.Number of theoretical plate presses Vitexin rhamnose
Glycosides peak, which calculates, should be not less than 2500.
The preparation precision of reference substance solution is weighed in dry 12 hours vitexin rhamnoside reference substances of vacuum drying apparatus
In right amount, add Diluted Alcohol be diluted to scale be configured to every 1mL containing 50 μ g vitexin rhamnoside reference substance solutions to get.
The preparation of test solution takes this product under weight differential item, finely ground, takes about 0.25g, accurately weighed, sets 50ml amount
In bottle, add Diluted Alcohol 40ml, be ultrasonically treated 30 minutes, let cool, be diluted to scale with Diluted Alcohol, shake up, filters, take subsequent filtrate
5ml is placed in 10mL volumetric flask, Diluted Alcohol is added to be diluted to scale, shake up to get.
Measuring method is accurate respectively to draw reference substance solution and each 20 μ L of test solution, injects hplc determination, i.e.,
.
Three batches of hawthorne leaf P.E yield are made respectively by above-mentioned technique and assay the results are shown in Table 1.
1 hawthorne leaf P.E yield of table and assay result
| Batch | Yield (%) | General flavone content (%) | Vitexin rhamnoside content (%) |
| 1 | 25.18 | 29.13 | 2.78 |
| 2 | 25.52 | 28.83 | 2.72 |
| 3 | 25.4 | 28.59 | 2.7 |
| It is average | 25.37 | 28.85 | 2.73 |
Embodiment 2: the preparation of Longstamen Onion Bulb, Rhizoma Polygonati extract
3.5 parts and 1.5 parts of rhizoma polygonati of Longstamen Onion Bulb are weighed, 10 times of amount water are added for the first time under the conditions of 100 DEG C and decoct 2 hours, the
Secondary that 8 times of amount water decoctions 1 hour are added, collecting decoction, filtrate is concentrated into 1: 1, and ethyl alcohol is added to make alcohol content 55%, is centrifuged, takes
Supernatant, filtering are distilled to recover ethyl alcohol, continue distillation and concentration to appropriate, dry, pulverize to get.
The assay of Longstamen Onion Bulb, Rhizoma Polygonati extract
The preparation of reference substance solution learns from else's experience 105 DEG C of dryings to the DEXTROSE ANHYDROUS reference substance 33mg of constant weight, accurately weighed, sets
It in 100ml measuring bottle, is dissolved in water and is diluted to scale, shake up to get (containing DEXTROSE ANHYDROUS 0.33mg) in every 1ml.
The preparation precision of standard curve measures reference substance solution 0.1ml, 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml,
It is set in 10ml plug test tube respectively, respectively adds water to 2.0ml, shaken up, 0.2% Anthrone Sulphuric acid is slowly added dropwise in ice-water bath
Solution to scale, mixing lets cool in postposition water-bath and keeps the temperature 10 minutes, takes out, and sets 10 minutes cooling in ice-water bath, taking-up immediately,
Using corresponding reagent as blank.According to UV-VIS spectrophotometry (" Chinese Pharmacopoeia " annex V A in 2010), in 582nm wave
Strong point measures absorbance, and using absorbance as ordinate, concentration is abscissa, draws standard curve.
Measuring method takes this product fine powder about 0.5g, accurately weighed, sets in round-bottomed flask, adds 80% ethyl alcohol 150ml, sets in water-bath
It being heated to reflux 1 hour, filters while hot, residue is washed 3 times with 80% hot ethanol, and each 10ml sets residue and filter paper in flask,
Adding water 150ml, sets and be heated to reflux in boiling water bath 1 hour, filtered while hot, residue and flask are washed 4 times, each 10ml with hot water,
Merging filtrate and washing lotion, let cool, and are transferred in 250ml measuring bottle, add water to scale, shake up, and precision measures 1ml, set 10ml tool plug
In dry test tube, the method under the preparation of sighting target directrix curve measures absorbance from " adding water to 2.0ml " in accordance with the law, from standard
On curve read test solution in the weight (mg) containing DEXTROSE ANHYDROUS, calculate to get.
Three batches of Longstamen Onion Bulbs, Rhizoma Polygonati extract yield and assay is made respectively by above-mentioned technique the results are shown in Table 2.
2 Longstamen Onion Bulb of table, Rhizoma Polygonati extract yield and assay result
| Batch | Yield (%) | Polyoses content (%) |
| 1 | 36.78 | 4.51 |
| 2 | 37.05 | 4.55 |
| 3 | 37.22 | 4.63 |
| It is average | 37.02 | 4.56 |
Embodiment 3: the preparation of composition tablet
Prescription
Preparation method: leaves of Hawthorn is extracted into obtain hawthorne leaf P.E according to 1 method of embodiment；Longstamen Onion Bulb, rhizoma polygonati are mentioned according to embodiment 2
Obtain Longstamen Onion Bulb, Rhizoma Polygonati extract.The auxiliary material of extract and recipe quantity is weighed, crushes, sieves with 100 mesh sieve respectively, it is spare.By leaves of Hawthorn
Extract, Longstamen Onion Bulb, Rhizoma Polygonati extract, starch, pregelatinized starch, microcrystalline cellulose are uniformly mixed, stir evenly, it is suitable for soft for being made
Material.Cross the pelleting of 18 meshes.Particle is dried under conditions of 60 DEG C.Sodium carboxymethyl starch and stearic acid is added in dried particle
Magnesium crosses 18 mesh sieves, is uniformly mixed.Sampling, semi-finished product chemical examination.According to the determining slice weight tabletting of chemical examination.Finished product full inspection, packaging
Storage.
Embodiment 4: the preparation of composition capsule
Prescription
Preparation method: leaves of Hawthorn is extracted into obtain hawthorne leaf P.E according to 1 method of embodiment；Longstamen Onion Bulb, rhizoma polygonati are mentioned according to embodiment 2
Obtain Longstamen Onion Bulb, Rhizoma Polygonati extract.The auxiliary material of extract and recipe quantity is weighed, crushes, sieves with 100 mesh sieve respectively, it is spare.By leaves of Hawthorn
Extract, Longstamen Onion Bulb, Rhizoma Polygonati extract, starch are uniformly mixed, stir evenly, suitable softwood is made.Cross the pelleting of 18 meshes.Particle
It is dried under conditions of 60 DEG C.18 mesh sieves are crossed, are uniformly mixed.Sampling, semi-finished product chemical examination.According to the determining weight dress of chemical examination
Capsule.Finished product full inspection, is packed and stored.
Embodiment 5: the preparation of composition granule
Prescription
Preparation method: leaves of Hawthorn is extracted into obtain hawthorne leaf P.E according to 1 method of embodiment；Longstamen Onion Bulb, rhizoma polygonati are mentioned according to embodiment 2
Obtain Longstamen Onion Bulb, Rhizoma Polygonati extract.The auxiliary material of extract and recipe quantity is weighed, crushes, sieves with 100 mesh sieve respectively, it is spare.By leaves of Hawthorn
Extract, Longstamen Onion Bulb, Rhizoma Polygonati extract, lactose powder are uniformly mixed in the method for equal increments, and 2%HPMC60% ethanol solution is added
In right amount, it stirs evenly, suitable softwood is made.Cross the pelleting of 18 meshes.Particle is dried under conditions of 55 DEG C.Dry particl crosses 18 mesh
Sieve whole grain.Sampling, semi-finished product chemically examine the content of main ingredient in particle, determine loading amount.Packaging, finished product full inspection are packed and stored.
Embodiment 6: the preparation of composition soft agent
Prescription
Preparation method: leaves of Hawthorn is extracted into obtain hawthorne leaf P.E according to 1 method of embodiment；Longstamen Onion Bulb, rhizoma polygonati are mentioned according to embodiment 2
Obtain Longstamen Onion Bulb, Rhizoma Polygonati extract.The auxiliary material of extract and recipe quantity is weighed, crushes, sieves with 100 mesh sieve respectively, it is spare.By recipe quantity
Soybean oil and soybean lecithin, beeswax heating melting, mix, let cool, be added hawthorne leaf P.E, Longstamen Onion Bulb, Rhizoma Polygonati extract are ground
It is even, it is pressed into soft capsule.
Embodiment 7: the preparation of haw thorn leaf total flavone extract
The hawthorne leaf P.E of Example 1 after the petroleum ether degreasing with 1/2 amount, discards petroleum ether liquid, then with acetic acid second
Ester extraction, extract liquor are recovered under reduced pressure ethyl acetate and are concentrated to dryness, and suitable quantity of water, which is added, to be made to dissolve, and are added on processed polyamide
On column (granularity: 30~60 mesh, with 95% ethyl alcohol wet method dress post, first with 95% ethanol elution of 3 times of column volumes, rear 3 times of cylinders
Long-pending water elution is spare to no alcohol taste), first with the water elution of 2 times of column volumes, water lotion is discarded, then with 3 times of column volumes
80% ethanol elution collects eluent, recycles ethyl alcohol and is concentrated into the concentrate of relative density about 1.02~1.08 (60 DEG C), does
It is dry, obtain haw thorn leaf total flavone extract.
Three batches of haw thorn leaf total flavone extract yields are made respectively by above-mentioned technique and assay the results are shown in Table 3.
3 haw thorn leaf total flavone extract yield of table and assay result
| Batch | Yield (%) | General flavone content (%) | Vitexin rhamnoside content (%) |
| 1 | 18.97 | 85.27 | 9.21 |
| 2 | 18.74 | 85.78 | 9.35 |
| 3 | 19.02 | 86.12 | 9.48 |
| It is average | 18.91 | 85.72 | 9.35 |
Embodiment 8: the preparation of Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract
Longstamen Onion Bulb, the Rhizoma Polygonati extract of Example 2 are repeated 4 times with 95% ethanol precipitation of diploid product.With
Small-molecule substance is fallen in dialysis membrane dialysis, again alcohol precipitation, finally with the anhydrous ether of equal volume and suitable acetone washing precipitating
Object, dry Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract.
Three batches of Longstamen Onion Bulbs, coarse solomon's seal polysaccharide extract yield and assay is made respectively by above-mentioned technique the results are shown in Table 4.
4 Longstamen Onion Bulb of table, coarse solomon's seal polysaccharide extract yield and assay result
| Batch | Yield (%) | Polyoses content (%) |
| 1 | 12.35 | 14.52 |
| 2 | 12.74 | 14.66 |
| 3 | 12.68 | 14.31 |
| It is average | 12.59 | 14.5 |
Embodiment 9: the preparation of composition liquid drugs injection
Prescription
Preparation method: haw thorn leaf total flavone extract and Longstamen Onion Bulb, rhizoma polygonati that the leaves of Hawthorn of recipe quantity is prepared according to embodiment 7 are weighed
Longstamen Onion Bulb, the coarse solomon's seal polysaccharide extract prepared according to embodiment 8.Pipeline and container etc. of the previous day processing with liquid are mentioned, use is faced
It is preceding to be rinsed again with fresh water for injection.Haw thorn leaf total flavone extract and Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract are added and match liquid
Completely, benefit adds to the full amount of water for injection for heating stirring dissolution in the water for injection of amount 70%.The needle with liquid measure 0.05% is added to use
Active carbon, heating stirring 15 minutes.Through sand stick filtering decarbonization.Measure and adjust the pH value of solution.Micropore filter through 0.45um
Film refined filtration.Check the clarity of solution, semi-finished product chemical examination.By solution encapsulating in glass ampule.100 DEG C of flowing steam sterilizations 30
Minute.Sample is put into while hot in 0.01% methylene blue solution and is hunted leak.Lamp inspection, finished product full inspection, is packed and stored.
Embodiment 10: the preparation of composition powder injection
Prescription
Preparation method: haw thorn leaf total flavone extract and Longstamen Onion Bulb, rhizoma polygonati that the leaves of Hawthorn of recipe quantity is prepared according to embodiment 7 are weighed
Longstamen Onion Bulb, the coarse solomon's seal polysaccharide extract prepared according to embodiment 8.First by container tool and antibiotic glass bottle with liquid, glue
Plug etc. carries out aseptic process.Haw thorn leaf total flavone extract and Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract are added with liquid measure 40%
Heating stirring dissolution is complete in sterile water for injection.Mannitol adds the sterile water for injection heating stirring of 30% with liquid measure to dissolve
Completely, merge above-mentioned solution, add sterile water for injection to full dose.The needle-use activated carbon for matching liquid measure 0.05% is added, heating is stirred
It mixes 15 minutes.Through sand stick filtering decarbonization.Measure and adjust the pH value of solution.Miillpore filter refined filtration through 0.22um.It checks molten
The clarity of liquid, semi-finished product chemical examination.It is sub-packed in antibiotic glass bottle, half tamponade.Sample is put into freeze dryer and is freeze-dried.
Be lyophilized by following lyophilized technique: 1. pre-freeze: being cooled to -35 DEG C, is kept for temperature 5 hours；2. low-temperature distillation: -35 DEG C of heat preservations,
It opens vacuum pump and vacuumizes holding 2 hours, then slowly heating, rises to 0 DEG C for temperature in 30 hours or so；3. high temperature drying: 2 hours
25 DEG C are inside warming up to, is kept the temperature to 2 hours；4. shutting down freeze-drying terminates.Lid is rolled in tamponade.Finished product full inspection, is packed and stored.
Embodiment 11: the preparation of composition sodium chloride injection
Prescription
Preparation method: haw thorn leaf total flavone extract and Longstamen Onion Bulb, rhizoma polygonati that the leaves of Hawthorn of recipe quantity is prepared according to embodiment 7 are weighed
Longstamen Onion Bulb, the coarse solomon's seal polysaccharide extract prepared according to embodiment 8.Pipeline and container etc. of the previous day processing with liquid are mentioned, use is faced
It is preceding to be rinsed again with fresh water for injection.Haw thorn leaf total flavone extract and Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract are added and match liquid
Sodium chloride completely, is used the water for injection with liquid measure 20% to dissolve complete by heating stirring dissolution in the water for injection of amount 40%.It closes
And above-mentioned solution, benefit add to the full amount of water for injection.Needle-use activated carbon of the addition with liquid measure 0.05%, heating stirring 15 minutes.Through
Sand stick filtering decarbonization.Measure and adjust the pH value of solution.Miillpore filter refined filtration through 0.45um.Check the clarity of solution,
Semi-finished product chemical examination.It is filling in the infusion bottle of 250ml.115 DEG C pressure sterilizing 30 minutes.Lamp inspection, finished product full inspection, is packed and stored.
Embodiment 12: the preparation of composition glucose injection
Prescription
Preparation method: haw thorn leaf total flavone extract and Longstamen Onion Bulb, rhizoma polygonati that the leaves of Hawthorn of recipe quantity is prepared according to embodiment 7 are weighed
Longstamen Onion Bulb, the coarse solomon's seal polysaccharide extract prepared according to embodiment 8.Pipeline and container etc. of the previous day processing with liquid are mentioned, use is faced
It is preceding to be rinsed again with fresh water for injection.Haw thorn leaf total flavone extract and Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract are added and match liquid
Glucose completely, is used the water for injection with liquid measure 20% to dissolve complete by heating stirring dissolution in the water for injection of amount 40%.It closes
And above-mentioned solution, benefit add to the full amount of water for injection.Needle-use activated carbon of the addition with liquid measure 0.05%, heating stirring 15 minutes.Through
Sand stick filtering decarbonization.Measure and adjust the pH value of solution.Miillpore filter refined filtration through 0.45um.Check the clarity of solution,
Semi-finished product chemical examination.It is filling in the infusion bottle of 250ml.115 DEG C pressure sterilizing 30 minutes.Lamp inspection, finished product full inspection, is packed and stored.
Test sample study on the stability
Composition capsule, prescription and preparation method referring to 4 capsule of embodiment preparation.
Investigation project: character, moisture, content.
Accelerated test and long-term stable experiment method and result: this product is set into 40 DEG C ± 2 DEG C of temperature, relative humidity 75%
It is placed under conditions of ± 5% and places 12 months under conditions of 6 months and 25 DEG C ± 2 DEG C of temperature, relative humidity 60% ± 10%, respectively
Index has no significant change, the experimental results showed that composition capsule place for a long time it is basicly stable.
Observation of curative effect
The research of 1 present composition acute toxicity test of test example
Kun Ming mice 20 are taken, half male and half female.Fasting 12 hours, every gavaged 40% (g/mL) by 0.4ml/10g
Present composition medicinal powder (is prepared) with purified water.Maximum concentration, maximum administered volume are reached.It is gavaged in one day 1 time, it is continuous to see
Mouse activity condition and The dead quantity in 7 days are examined, after as a result mouse presses above-mentioned dosage, without death, activity in 7 days
Freely, diet is normal, and hair is glossy, no loose stools etc..The maximal tolerance dose for being computed mouse is 16g/Kg, for clinical adult dosage
356 times.
The research of 2 present composition general pharmacology of test example
1) to the influence of Kunming mouse general behavior performance
After gastric infusion, for each administration group animal compared with blank group, activity is normal, walks with a steady step, is existing without salivation, amyostasia etc.
As having no generation adverse reaction.
2) to the influence of Kunming mouse spontaneous activity
0.5h sets animal in mouse activity count instrument active box after gastric infusion, records in 10min after adapting to 5min
Animal activity number；Each dosage group animal activity number compared with blank group.As a result be administered after groups of animals compared with blank group,
Number of activities do not occur significant difference, and data are shown in Table 5.
Influence of 5 drug of table to mouse autonomic activities
| Group | Dosage (mg/kg) | Number of animals | Number of activities |
| Blank group | - | 10 | 236.8±21.76 |
| High dose group | 2450 | 10 | 233.2±15.68 |
| Middle dose group | 1220 | 10 | 239.9±18.85 |
| Low dose group | 409 | 10 | 231.4±18.88 |
Note: * P < 0.05 (compared with model group).
3) influence that Kunming mouse is fallen asleep is induced to sub-threshold dose yellow Jackets
1h after groups of animals gastric infusion, intraperitoneal injection yellow Jackets 32mg/kg (for measured in advance animal 90%~
The maximum threshold dose that 100% mouse righting reflex does not disappear), righting reflex disappears within 15min after record yellow Jackets injection
Lose the mouse number of 1min or more.As a result: each group experimental animal induces sub-threshold dose yellow Jackets after giving corresponding drug
Mouse, which falls asleep, does not detect difference, and display sub-threshold dose yellow Jackets induction mouse sleep is uninfluenced, and data are shown in Table 6.
Table 6 induces sub-threshold dose yellow Jackets the influence that mouse falls asleep
| Group | Dosage (mg/kg) | Number of animals | Sleep number of elements | Male and female | Sleep rate (%) |
| Blank group | - | 10 | 6 | ♀2♂4 | 60 |
| High dose group | 2450 | 10 | 8 | ♀5♂3 | 80 |
| Middle dose group | 1220 | 10 | 6 | ♀3♂3 | 60 |
| Low dose group | 409 | 10 | 7 | ♀5♂2 | 70 |
4) to the influence of yellow Jackets sleeping time
Yellow Jackets 40mg/kg is injected intraperitoneally (for measured in advance to 1h after gastric infusion in the stomach-filling of each group Kunming mouse
Animal 100% fall asleep minimum dose), using righting reflex loss as time for falling asleep, from righting reflex loss to recovery time
For sleep time (min).As a result for groups of animals compared with blank group, sleeping time does not occur significant difference after being administered,
Data are shown in Table 7.
Influence of the table 7 to pentobarbital sodium in mice sleeping time
| Group | Dosage (mg/kg) | Number of animals | Sleep number of elements |
| Blank group | - | 10 | 30.73±1.35 |
| High dose group | 2450 | 10 | 31.45±2.26 |
| Middle dose group | 1220 | 10 | 29.13±2.77 |
| Low dose group | 409 | 10 | 30.66±2.41 |
Note: * P < 0.05 (compared with model group).
5) to the influence of the mouse coordinated movement of various economic factors
Surface is taken to twine white glue cloth, diameter 1cm, long 60cm iron staff are vertical fixed.1h after gastric infusion, mouse is placed in
Top makes it creep naturally, observes mouse coordinated movement of various economic factors ability, and record it and climb to the bottom end time.As a result groups of animals and sky
White group is compared, and is creeped steadily, does not slide and fall phenomenon, and the pole-climbing time does not occur significant difference, and data are shown in Table 8.
Influence of the table 8 to the mouse pole-climbing time
| Group | Dosage (mg/kg) | Number of animals | The pole-climbing time (s) |
| Blank group | - | 10 | 11.88±0.60 |
| High dose group | 2450 | 10 | 12.25±0.48 |
| Middle dose group | 1220 | 10 | 12.23±0.52 |
| Low dose group | 409 | 10 | 11.56±0.83 |
Note: * P < 0.05 (compared with model group).
6) to the influence of Wistar Rat Cardiovascular system
With yellow Jackets intraperitoneal injection of anesthesia animal, separates arteria carotis communis and connect with System of organism signal
Recording blood pressure and heart rate；Electrocardiogram is traced with limb leads (II lead).Record is primary respectively before gastric infusion, gives then at stomach-filling
These parameters are recorded after medicine.As a result compared with blank group, electrocardio and blood pressure items numerical value significantly groups of animals do not occur after being administered
Sex differernce, data are shown in Table 9 and table 10.
Table 9 is on the cardiac electrical influence of rat
| Group | Dosage (mg/kg) | Number of animals | Heart rate (beat/min) | Maximum value (mV) | Minimum value (mV) |
| Blank group | - | 8 | 367.53±21.34 | 0.41±0.08 | -0.26±0.10 |
| High dose group | 1701 | 8 | 365.70±14.04 | 0.47±0.10 | -0.28±0.14 |
| Middle dose group | 850.5 | 8 | 365.39±43.58 | 0.48±0.03 | -0.32±0.16 |
| Low dose group | 283.5 | 8 | 365.24±56.27 | 0.39±0.07 | -0.28±0.19 |
Note: * P < 0.05 (compared with model group).
Influence of the table 10 to rat blood pressure
| Group | Dosage (mg/kg) | Number of animals | Systolic pressure (mmHg) | Diastolic pressure (mmHg) |
| Blank group | - | 8 | 72.95±12.74 | 45.52±13.23 |
| High dose group | 1701 | 8 | 68.92±9.33 | 44.54±9.18 |
| Middle dose group | 850.5 | 8 | 69.80±21.32 | 44.44±21.32 |
| Low dose group | 283.5 | 8 | 86.91±17.36 | 59.61±15.44 |
Note: * P < 0.05 (compared with model group).
7) to the influence of Wistar rats respiratory system
Respiratory rate, average expiration peak pressure and the average suction paddy pressure of System of organism signal record animal.Knot
For groups of animals compared with blank group, respiratory rate, expiration peak value and air-breathing valley do not occur significant difference after fruit administration, number
According to being shown in Table 11.
Influence of the table 11 to respiratory system in rats
| Group | Dosage (mg/kg) | Number of animals | Frequency (beat/min) | Maximum value (ml/s) | Minimum value (ml/s) |
| Blank group | - | 8 | 123.20±12.11 | 0.13±0.14 | 0.02±0.14 |
| High dose group | 1701 | 8 | 119.35±6.85 | 0.24±0.13 | 0.11±0.13 |
| Middle dose group | 850.5 | 8 | 117.49±17.16 | 0.24±0.10 | 0.12±0.11 |
| Low dose group | 283.5 | 8 | 113.60±1.78 | 0.26±0.17 | 0.17±0.19 |
Note: * P < 0.05 (compared with model group).
For the present composition in the detection of general pharmacology indices, each administration group has no significant compared with blank group
Sex differernce.Illustrate the present composition to nervous system, cardiovascular and respiratory system does not generate adverse effect, can be applied to face
Bed.
The research of 3 present composition pharmacodynamics of test example-composition treatment rats with myocardial ischemia pharmacodynamics test.
Myocardial infarction model is replicated according to literature method.With 1% yellow Jackets (0.04g/kg) using intraperitoneal injection fiber crops
Liquor-saturated, dorsal position is fixed on operating table.Record standard II lead electrocardiogram, row tracheotomy are connected with breathing apparatus, open
Chest exposes heart, passes through the left room branch of coronary artery, ligation arteria coroaria sinistra (artificial hand with surgical thread between aorta circular cone and left auricle of heart
Art group, which only threads, not to ligature), heart resets, and closes thoracic cavity after drum lung rapidly, and penicillin is embrocated in wound, prevents infected wound.
Close chest after observe rats breathing recovery situation, if any autonomous respiration can off line put back to mouse cage.There is the back of a bow with ECG ST section
It raises as myocardial ischemia modeling success.Chronic myocardial ischemia model is formed after four weeks.
1) to the influence of rats with myocardial ischemia S-T segment after coronary artery ligation
Rats with myocardial ischemia after screening is uniformly divided into 5 groups at random in addition to sham-operation group, sham-operation group, model group, sun
Property medicine control group, high dose group, middle dose group, 6 groups of low dose group, 72, are raised totally together by every group 12.Administration group stomach-filling
That gives corresponding dosage contains drug solns, and blank group, model group stomach-filling give the distilled water of equivalent, once a day, totally 10 days.Last
Each group rat 1% yellow Jackets (40mg/kg) intraperitoneal anesthesia after administration connects biological functional system, monitors limbs
Lead electrocardiogram measures S-T segment.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group and model group
S-T segment can be improved by, which comparing, raises, and data are shown in Table 12.
The influence that table 12 changes ECG ST section
| Group | Number of animals (only) | Dosage (mg/kg) | ST sections of changes (mv) |
| Blank group | 12 | - | 0.07±0.03 |
| Model group | 12 | - | 0.23±0.02* |
| High dose group | 12 | 1701 | 0.12±0.02# |
| Middle dose group | 12 | 850.5 | 0.12±0.02# |
| Low dose group | 12 | 283.5 | 0.18±0.01# |
| Positive control | 12 | 324 | 0.12±0.02# |
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
2) to the influence of Serum fibrosis markers
Experimental animal grouping and administration are same as above.In last dose 1 hour, eyeball took blood, operated by kit specification, and
Serum creatine kinase (CK), lactic dehydrogenase (LDH), aspartate amino transferase are detected with semi-automatic biochemical analyzer
(AST) active, record data.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group and model group ratio
Compared with the present composition can be substantially reduced creatine kinase in serum (CK), lactic dehydrogenase (LDH), Aspartate amino and turn
It moves enzyme (AST), data are shown in Table 13.
Influence of the table 13 to Serum fibrosis markers
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
3) to the influence of myocardial metabolism product
Experimental animal grouping and administration are same as above.In last dose 1 hour, eyeball took blood, operated by kit specification, and
Serum lactic (LD), free fatty acid (NEFA), Na-KATP enzyme index are detected with 756PC type ultraviolet-uisible spectrophotometer.Note
Record data.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group is compared with model group group, the present invention
Composition atpase activity reduces, and lactic acid and free fatty acid content reduce, and data are shown in Table 14.
Influence of the table 14 to myocardial metabolism product index
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
4) to the influence of Plasma free radical damage criterion
Experimental animal grouping and administration are same as above.In last dose 1 hour, eyeball took blood, operated by kit specification, and
Malonaldehyde (MDA), total number born (SOD), glutathione peroxide are detected with 756PC type ultraviolet-uisible spectrophotometer
Compound enzyme (GSH-Px) index.Record data.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group
Compared with model group, each administration group of the present composition can be such that malonaldehyde (MDA) content reduces, total number born
(SOD), the activity of glutathione peroxidase (GSH-Px) increases, and data are shown in Table 15.
Influence of the table 15 to myocardial metabolism product index
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
5) to the influence of hemorheology index
Experimental animal grouping and administration are same as above.In last dose 1 hour, abdominal aortic blood sent and examines with hospital laboratory
It tests.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group is compared with model group, the present composition
Each administration group whole blood viscosity reduces, plasma viscosity reduces, hematocrit reduces, erythrocyte sedimentation rate reduces, and illustrates that it can inhibit red blood cell
Aggregation reduces erythrocyte fragility, enhances its morphotropism, be shown in Table 16.
6) to the influence of hemodynamic index
Experimental animal grouping and administration are same as above.In last dose 1 hour, rat is lain on the back and is fixed on operating table, cut
Skin of neck, separates right carotid, and row arterial cannulation is moved right side neck is inserted into containing the conduit of 0.1% heparin sodium liquid
Arteries and veins is inserted into left ventricle through right common carotid artery, judges whether intubation enters ventricle according to the change of pressure figure shown in display,
Cardiac catheter connects pressure transducer, inputs a signal into physiograph.And recorded heart rate (HR), left ventricular systolic pressure
(LVSP), maximal ascending rate of internal pressure of left ventricle (+dp/dtmax), left ventricular diastolic pressure (LVDp), ventricular end diastolic pressure
Power (LVDEp), maximal descending rate of internal (- dP/dtmax).Record data.Comparison among groups are examined with t and are indicated, as a result
WithIt indicates.As a result each administration group is compared with model group, each administration group of the present composition can make heart rate tend to it is normal,
Left ventricular systolic pressure increases, left room diastolic pressure reduces, and left room maximum climbing speed increases, left room maximum fall off rate increases, terminal
Diastolic pressure reduces, and is shown in Table 17.
7) on the morphologic influence of myocardial histopathology
Experimental animal grouping and administration are same as above.In last dose 1 hour, heart is taken out, send and is examined in hospital pathology department.Knot
Fruit is shown in Table 18.
8) to myocardial necrosis area estimation
Experimental animal grouping and administration are same as above.In last dose 1 hour, rat is put to death, manubrium is fixed with tweezers, cuts off
Breastbone entirely cuts heart, removes blood stains with the normal saline flushing of pre-cooling, and filter paper suck dry moisture is whole-heartedly weighed, and -30 DEG C
Frost half an hour is cut into the myocardium piece of 1~2mm thickness in parallel along left room long axis, is uniformly cut into 4~5 under heart ligature,
It being placed in the TTC solution of 1% concentration (PBS buffer solution of pH value 7.4), 37 DEG C of temperature incubate 15min, it is rinsed with water extra dyestuff,
Filter paper suck dry moisture, it is seen that normal myocardium is red, and ischemic myocardium is canescence, cuts off infarcted myocardium, is weighed, and infarct model is calculated
Enclose (infarcted region accounts for whole-heartedly heavy percentage).Record data, comparison among groups with t examine indicate, as a result with It indicates.As a result
Each administration group is compared with model group, and each administration group of the present composition can be such that myocardial infarction area significantly reduces, and difference has aobvious
Work property, is shown in Table 19.
Influence of the table 16 to hemorheology
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
Influence of the table 17 to hemorheology
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
Table 18 is on the morphologic influence of myocardial histopathology
Influence of the table 19 to myocardial infarction area
| Group | Number of animals (only) | Dosage (mg/kg) | Myocardial infarction area (%) |
| Model group | 12 | - | 13.01±0.63 |
| High dose group | 12 | 1701 | 7.00±0.83# |
| Middle dose group | 12 | 850.5 | 5.89±0.39# |
| Low dose group | 12 | 283.5 | 9.50±1.19# |
| Positive control | 12 | 324 | 7.08±0.74# |
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
Conclusion (of pressure testing): the improvement and treatment that the present composition has the change of myocardial infarction and ischemia model rat indices
Effect.
The research of 4 present composition pharmacodynamics of test example-composition treatment hyperlipidemia rats pharmacodynamics test.
1) to the influence of hyperlipidemia rats lipid-metabolism level
High lipid food ingredient and preparation: 3% cholesterol, 10% lard, 5% dried hen egg yolk, 0.3% sodium taurocholate, 0.2% first
Base thiouracil, 81.5% basal feed.10kg high lipid food adds 1~2kg white sugar.In proportion first by cholesterol, dried hen egg yolk,
Then sodium taurocholate, propylthiouracil, white sugar ground and mixed are added in the lard of warm, then with basal feed mix thoroughly to get.
Modeling: 72 SD rat normal diets are fed one week.Random selection 12 is only used as blank group mouse, and weighing label is given
Give normal diet nursing.Remaining 60 mouse weighing label, gives high lipid food nursing, it is high to feed progress in 4 weeks altogether for equal free water
The foundation of pionemia model.Record starts the rat body weight of surrounding after modeling respectively, and takes blood in 4th week end eyeground vein clump
1.5~2ml is stood, 3500rmin in centrifuge tube-1It is centrifuged 15min, serum is separated, is measured with semi-automatic biochemical analyzer
Total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein in its Diagnostic Value of Fasting Serum
The content of cholesterol (HDL-C), detection method in accordance with corresponding reagent box specification.Total cholesterol (TC), glycerol three in serum
When ester (TG) content is obviously higher than blank group, illustrate that hyperlipidemia animal model is successfully prepared.Next day starts gastric infusion, positive
For medicine with 0.324g/kg gastric infusion, the present composition is high, in, low dose group respectively with 1.701g/kg, 0.8505g/kg,
0.2835g/kg gastric infusion, totally 10 days.Continue to give high lipid food during administration.After last dose 1 hour, socket of the eye internal jugular vein
Clump takes blood 2.5ml, 4 DEG C of standings, 3500rmin-1It is centrifuged 15min, separates serum, in accordance with corresponding reagent box specification, certainly with half
Automatic Biochemical Analyzer detects total cholesterol (TC), triglycerides (TG) level and Apolipoprotein A1 (ApoA1) and apolipoprotein B
(ApoB) horizontal.Comparison among groups are examined with t and are indicated, are as a result indicated with X ± S.As a result each administration group can be bright compared with model group
It is aobvious to reduce total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-C) and apolipoprotein B in serum
(ApoB) content, hence it is evident that increase high-density lipoprotein cholesterol (HDL-C) and Apolipoprotein A1 (ApoA1) content, be shown in Table 20.
Influence of the table 20 to hyperlipidemia lipid-metabolism level
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
2) to the influence of hyperlipidemia rats level of lipid
Experimental animal grouping and administration are same as above.In last dose 1 hour, socket of the eye internal jugular vein clump took blood, 4 DEG C of standings, 3500r
min-1It is centrifuged 15min, separates serum, in accordance with corresponding reagent box specification, detects total super oxygen with ultraviolet-uisible spectrophotometer
Object mutase (SOD), malonaldehyde (MDA), nitric oxide (NO) and glutathione peroxidase (GSH-Px) are horizontal.Compare between group
Compared with t inspection indicate, as a result withIt indicates.As a result each administration group can significantly reduce malonaldehyde (MDA) compared with model group
Content, hence it is evident that increase the content of nitric oxide (NO), increase total number born (SOD), glutathione peroxidase
(GSH-Px) active, it is shown in Table 21.
Influence of the table 21 to hyperlipidemia level of lipid
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
3) to the influence of hyperlipidemia rats hemorheology index
Experimental animal grouping and administration are same as above.In last dose 1 hour, abdominal aortic blood sent and examines with hospital laboratory
It tests.Comparison among groups with t examine indicate, as a result withIt indicates.As a result each administration group compared with model group whole blood viscosity reduce,
Plasma viscosity reduces, hematocrit reduces, erythrocyte sedimentation rate reduces, and illustrates that it can inhibit erythrocyte aggregation, reduces erythrocyte fragility, increases
Its strong morphotropism, is shown in Table 22.
Influence of the table 22 to hyperlipidemia lectin from hemolymph
Note: compared with blank group, * P < 0.05；Compared with model group, #P < 0.05.
4) to the influence of hyperlipidemia rats liver organization pathomorphism
Experimental animal grouping and administration are same as above.After last dose 1h, conventional materials, 10% formalin are carried out to hepatic tissue
Solution is fixed, conventional H E dyeing.Optical microphotograph microscopic observation liver organization form.This send examines in hospital pathology department.As a result
It is shown in Table 23.
Table 23 is on the morphologic influence of myocardial histopathology
Conclusion (of pressure testing): the improvement and therapeutic effect having to the change of Hyperlipemia model rat indices.
The research of 5 present composition long term toxicity test of test example.
Experiment purpose: observation repeats the orally administration present composition to toxic reaction caused by rat, the disease of appearance
Shape and severity, and the target organ of toxicity and its degree of reversibility of damage are provided, nontoxic crude protein is determined, with evaluation
The safety of present composition long-term administration provides reference to draft clinical test dosage and observation index.
Experimental animal: detergent SD big white mouse, half male and half female, quantity 120,80 grams~110 grams of weight (6~9 week old).
Animal packet: animal is normally raised 1 week in laboratory first, rejects sick mouse and abnormal mouse, takes rat 120, press
Weight is randomly divided into 4 groups, and every group 30, half male and half female.Respectively Normal group, high, medium and low three agent of the present composition
Amount group.
Dosage: the maximal tolerance dose of mouse is 16g/Kg, does not occur toxic reaction, therefore, in the case, according to
The administration of rat high dose group should be greater than 50 times clinical or more principle design dosage.Present composition high dose group rat
(60 times of quasi- dosage of clinic) is administered by 2.7g/kg, middle dose group rat is by (the 30 times of quasi- agent of clinic of 1.35g/kg gastric infusion
Amount), low dose group rat presses 0.45g/kg gastric infusion (10 times of quasi- dosage of clinic), volume 2ml/100g, normal control
Group gives same volume distilled water.
Administration time: it is administered once on every Mondays to the morning 8:30 of Saturday or so, Sunday is not administered.Successive administration 6 months.
Observation index
1) general Symptom Observation
1. observing time: being observed before administration with every morning convalescence each cage, the upper and lower noon is observed simultaneously during administration.
2. observing number of cases: all buying animal before administration, total Test animal after administration.
3. observing frequency: terminating from day to convalescence is bought, once a day.
4. observed content: observing appearance, coat, the gait, behavioral activity of animal, the reaction to sound has atremia, convulsion
Contraction, whether breathing is steady, there have to be without exception.When administration and after administration, in addition to paying attention to observing above-mentioned reaction, also especially to observe whether there is or not
Drowsiness, vomiting, excrement shape and color etc..Administration will also observe reactivity when arresting animal simultaneously, eye, nose, around mouth
State, whether there is or not manure contaminations at lower abdomen, anus, and skin color, whether there is or not wound, tumour etc., the state of thorax abdomen, muscle tonues
Degree etc..
2) body weight determination
1. measuring method: being measured using electronic balance.
2. measuring number of cases: all buying animal before administration, total Test animal after administration.
3. measuring frequency: buying day and grouping day (before administration) is respectively surveyed once, during administration and convalescence claims weekly one
Secondary weight, in addition, first measuring the weight of animal in progress when dissected.
3) food ration measures
1. measuring method: it is every group of daily food ration that every group, which has supply altogether to subtract surplus altogether, and every group is taken the photograph daily
Appetite is every group of daily food ration of every animal divided by every group of number of animals.
2. measuring number: measurement is primary before administration, measures weekly once during administration with convalescence.
3) hematology and blood biochemical analysis detection
1. detecting period: it is primary to terminate each detection in administration 3 months, 6 months (administration terminates) and convalescence.
2. detecting number of cases: administration 3 months, administration 6 months (administration terminates) and convalescence terminate each group and detect 10 respectively.
3. blood-sampling method: Rat Fast 14 hours or more before taking a blood sample, urethane 0.1g/100g weight intraperitoneal injection of anesthesia,
Special blood-drawing pipe and blood taking needle are taken a blood sample from abdominal aorta.Peripheral blood cell counts project and method are shown in Table 24.
24 peripheral blood cell counts project of table and method
| Project | It is anticoagulant | Measuring method | Use instrument |
| Red blood cell count(RBC) (RBC) | EDTA-K2 | Electric-resistivity method | Animal blood analyzer ( |
| Hemoglobin (HGB) | EDTA-K2 | Photoelectric colorimetry | Animal blood analyzer ( |
| Hematid specific volume (HCT) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Mean corpuscular volume (MCV) (MCV) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Average hemoglobin amount (MCH) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Mean hemoglobin concentration (MCHC) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| White blood cell count(WBC) (WBC) | EDTA-K2 | Electric-resistivity method | Animal blood analyzer ( |
| Neutrophil leucocyte percentage (GRAN%) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Cent lymphocytes (LYMPH%) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Monocyte percentage (MONO%) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
| Platelet count (PLT) | EDTA-K2 | Histogram calculation method | Animal blood analyzer ( |
Note: when discovery has an impact to hemopoietic system, the inspection of further progress marrow is answered.
4. biochemistry detection blood sample treatments: after abdominal aorta blood sampling, it is stored at room temperature 1 hour or so, centrifugation 3000rpm,
10min separates serum.Serum cannot such as detect on the same day, and serum is put into cryopreservation tube, and -80 DEG C freeze.Specific detection project and
Method is referring to current edition novel technique guideline.Specific detection project and method are shown in Table 25.
25 blood parameters detection project of table and method
| Project | Measuring method | Use instrument |
| Glutamic-pyruvic transaminase (ALT) | Continuous monitoring method | Automatic clinical chemistry analyzer |
| Glutamic-oxalacetic transaminease (AST) | Continuous monitoring method | Automatic clinical chemistry analyzer |
| Alkaline phosphatase (ALP) | Continuous monitoring method | Automatic clinical chemistry analyzer |
| Cretinephosphokinase (CK) | Continuous monitoring method | Automatic clinical chemistry analyzer |
| Urea nitrogen (BUN) | Dynamic method | Automatic clinical chemistry analyzer |
| Creatinine (CREA) | Dynamic method | Automatic clinical chemistry analyzer |
| Total protein (TP) | End-point method | Automatic clinical chemistry analyzer |
| Albumin (ALB) | End-point method | Automatic clinical chemistry analyzer |
| Globulin (GIB) | End-point method | Automatic clinical chemistry analyzer |
| Blood glucose (GLU) | End-point method | Automatic clinical chemistry analyzer |
| Total bilirubin (TBIL) | End-point method | Automatic clinical chemistry analyzer |
| Total cholesterol (CHOL) | End-point method | Automatic clinical chemistry analyzer |
| Triglyceride (TG) | End-point method | Automatic clinical chemistry analyzer |
| Potassium ion (K+) concentration | Electrode method | Blomelicalbloodgasandelectrolrteanalyzers |
| Sodium ion (Na+) concentration | Electrode method | Blomelicalbloodgasandelectrolrteanalyzers |
| Chloride ion (Cl-) concentration | Electrode method | Blomelicalbloodgasandelectrolrteanalyzers |
5) dead or moribund animals processing
1. processing requirement: finding as early as possible, dissect in time, seek reason as possible.
2. moribund animals: recording dying time, weight, symptom.
3. dead animal: the time of the dead discovery of record, weight, postmortem finding of naked eye.
4. biochemical analysis: moribund animals take blood to carry out hematology and blood biochemistry detection before putting to death.
5. pathologic finding: in addition to confirming animal dead due to caused by stomach-filling, other dying and dead animal be intended into
Row histopathologic examination.
6) it dissects
1. anatomic method: being anaesthetized with the intraperitoneal injection of urethane 0.1g/100g weight.Before dissect, to each animal
Situation is checked comprehensively；When dissected is carried out one by one by system, is had between observation internal organs color, form, positional relationship and internal organs
Nothing is adhered；Whether there is or not extravasated blood, hyperemia, blutpunkte, oedema, ulcer for organ surface and digest tube inner membrance；Thoracic cavity, abdominal cavity, cavum pericardiale have
Without pathological changes such as hydrops, such as note abnormalities, its position of accurate recording and pathological change in the remarks column of the record seen in dissection
Type.
2. dissecting the time: same to detection time
3. dissecting number of cases: administration 3 months, administration 6 months (administration terminates) and 1 month convalescence end each component do not dissect
10.
7) organ weights measure
Measuring method: the absolute weight for weighing 13 internal organs (following table) respectively using electronic balance (while calculating internal organs system
Number).
1. measuring period: when dissected measurement
2. measuring number of cases: identical with dissection number of cases
3. internal organs table:
| Serial number | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| Internal organs | Brain (brain, cerebellum) | Heart | Liver | Spleen | Lungs | Kidney | Adrenal gland |
| Internal organs | Thymus gland | Uterus | Ovary | Testis | Epididymis | Prostate |
8) histopathologic examination
All animals press current edition novel technique guideline and take out required organs and tissues, and visually observe and think
Abnormal tissue and internal organs.Sample is fixed in 10% formalin fixer one week, is repaired block and is fixed 12 hours again later, group
(decalcification of elder generation, skeletal tissue) is knitted with gradient alcohol dehydration, dimethylbenzene is transparent, paraffin embedding, paraffin slicing machine slice, H.E. dyeing,
Neutral gum sealing, light microscopy checking.Record the type and degree of lesion.As pathological change, other dosage groups occur for a certain tissue
The animal tissue should also carry out histopathological examination to determine dose-response relationship.
Observe organs and tissues table:
Experimental result
1) ordinary circumstance: for groups of animals during administration and surrounding convalescence, appearance signs, behavioral activities are normal, urine, excrement
For character without abnormal, diet is normal, and groups of animals weight increases and decreases without significant difference, and group difference is unobvious.Food-intake is shown in Table 26, table
27 and table 28.Influence to weight the results are shown in Table 29, table 30 and table 31.
26 medication of table, 3 months rats eating amounts (g/ days /)
27 medication of table, 3~6 months rats eating amounts (g/ days /)
The variation of table 28 convalescence rats eating amount (g/ days/only)
2) hematological indices inspection result
24 hours after rat is administered 90 days and 180 days, each group rat each 10, anticoagulated whole is collected in abdominal aorta blood sampling
Blood detects hematology indices with blood counting instrument.24 hours after last time is administered, the work of each group rat is deposited after killing 10
10 rats stayed stop administration, normal to raise, and continuing observation surrounding, all work is killed again.The case where when with medication 6 months, is complete
Under conditions of exactly the same, the index of detection is also identical.
Administration 90 days after the result shows that: compared with blank control group, high dose group Erythrocytes (RBC) significantly reduce,
Average volume of red blood cells (MCV) significantly increases, and monocyte percentage (MONO%) is remarkably decreased；Low dose group lymphocyte hundred
Divide and is remarkably decreased than (LYMPH%).Remaining index: such as hematocrit (HCT), average hemoglobin amount (MCH), average blood red
Protein concentration (MCHC), platelet count (PLT), hemoglobin (HGB) content, total white blood cells (WBC) and its neutrophil(e) granule
Cell percentages (GRAN%) differential counting, for each administration group compared with blank control group, no significant difference the results are shown in Table 32 and table
33。
Variation to rat blood routine in 32 medication of table 3 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| WBC(×109/L) | 11.62±2.34 | 13.37±4.97 | 13.00±3.87 | 9.69±1.84 |
| RBC(×1012/L) | 8.69±1.07 | 7.33±1.05* | 8.65±1.00 | 8.40±1.45 |
| HGB(g/L) | 152.00±13.42 | 135.50±21.59 | 147.70±13.62 | 146.50±17.44 |
| HCT (%) | 42.07±3.95 | 37.87±5.80 | 41.56±4.78 | 40.90±6.86 |
| MCV(fL) | 48.61±2.70 | 51.62±2.34* | 48.09±1.96 | 48.73±1.51 |
| MCH(pg) | 17.57±0.98 | 18.44±0.88 | 17.12±0.88 | 17.60±1.37 |
| MCHC(g/L) | 361.70±14.29 | 357.50±19.48 | 355.60±18.97 | 361.50±26.56 |
| PLT(×109/L) | 545.30±59.03 | 570.20±111.71 | 564.00±72.03 | 542.00±84.41 |
Note: compared with blank control group, * P < 0.05.
Variation to rat WBC differential counting (%) in 33 medication of table 3 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| GRAN% | 19.38±9.39 | 29.12±20.01 | 23.94±12.39 | 23.36±13.18 |
| LYMPH% | 58.71±10.18 | 48.46±21.78 | 47.94±18.71 | 40.51±21.65* |
| MONO% | 21.91±1.99 | 12.42±6.47*** | 18.12±7.66 | 16.13±9.05 |
Note: compared with blank control group, * P < 0.05；* * P < 0.001.
After administration 180 days the result shows that: compared with blank control group, the obvious drop of high and low dose group leucocyte (WBC) sum
Low (P < 0.05), wherein high dose group reduces most obvious；In, low dose group monocyte percentage (MONO%) significantly increase；
High, medium and low dosage group red blood cell (RBC) sum, hemoglobin (HGB) content and hematocrit (HCT) significantly reduce,
In: in, low dose group reduce it is most obvious；Middle dose group average hemoglobin amount (MCH) and mean hemoglobin concentration (MCHC)
It is significantly raised；Remaining index: such as average volume of red blood cells (MCV), platelet count (PLT), total white blood cells (WBC) and its point
Class counts [cent lymphocytes (LYMPH%) and neutrophil cell percentage (GRAN%)], each administration group and blank pair
Compare according to group, no significant difference between group the results are shown in Table 34 and table 35.
Variation to rat blood routine in 34 medication of table 6 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| WBC(×109/L) | 14.65±2.66 | 8.88±2.79*** | 12.42±4.08 | 10.75±3.52* |
| RBC(×1012/L) | 10.62±1.46 | 8.55±1.70** | 8.33±0.68*** | 8.08±0.71*** |
| HGB(g/L) | 176.90±14.98 | 97.80±68.76** | 152.80±6.76*** | 126.00±25.14*** |
| HCT (%) | 52.10±4.87 | 42.35±8.24** | 40.61±3.54* | 39.52±2.45*** |
| MCV(fL) | 49.19±2.78 | 49.59±2.23 | 48.78±2.78 | 49.11±3.84 |
| MCH(pg) | 16.98±1.29 | 13.35±6.30 | 18.41±1.29* | 15.63±3.14 |
| MCHC(g/L) | 339.60±139.90 | 266.38±121.87 | 377.5.00±19.58*** | 319.10±62.71 |
| PLT(×109/L) | 843.80±141.31 | 726.89±135.95 | 854.00±110.13 | 813.90±150.71 |
Note: compared with blank control group, * P < 0.05；* P < 0.01；* * P < 0.001.
Surrounding convalescence the result shows that: each administration group animal hematology index is compared with blank control group, without obvious between group
Difference the results are shown in Table 36 and table 37.
Variation to rat WBC differential counting (%) in 35 medication of table 6 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| GRAN% | 22.38±10.95 | 16.14±12.11 | 22.31±7.73 | 21.01±11.05 |
| LYMPH% | 62.21±17.39 | 61.20±23.35 | 61.89±8.16 | 63.94±11.09 |
| MONO% | 12.41±2.22 | 12.66±5.29 | 15.80±3.02* | 15.05±2.58* |
Note: compared with blank control group, * P < 0.05.
The variation of table 36 convalescence rat blood routine
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| WBC(×109/L) | 14.29±4.99 | 10.19±4.14* | 13.14±3.94 | 11.97±2.87 |
| RBC(×1012/L) | 9.04±1.24 | 8.50±0.59 | 8.73±0.70 | 8.48±1.15 |
| HGB(g/L) | 164.4±15.06 | 156.7±6.82 | 159.00±7.39 | 160.30±15.75 |
| HCT (%) | 43.45±4.41 | 41.88±2.16 | 42.43±2.55 | 41.70±4.64 |
| MCV(fL) | 48.27±2.48 | 49.40±3.13 | 48.72±1.97 | 49.38±3.11 |
| MCH(pg) | 18.29±1.26 | 18.50±1.39 | 18.27±0.82 | 19.05±1.61 |
| MCHC(g/L) | 378.70±10.18 | 374.40±11.25 | 375.10±9.97 | 385.20±12.60 |
| PLT(×109/L) | 820.80±86.69 | 749.80±145.39 | 819.80±165.79 | 794.10±120.85 |
Note: compared with blank control group, * P < 0.05.
The variation of table 37 convalescence rat WBC differential counting (%)
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| GRAN% | 26.90±12.30 | 31.83±10.98 | 22.33±9.37 | 23.07±10.12 |
| LYMPH% | 61.08±11.33 | 55.85±11.66 | 65.29±8.75 | 63.88±9.78 |
| MONO% | 12.02±3.24 | 12.32±1.73 | 12.33±1.34 | 13.04±1.57 |
3) blood parameters testing result
10 rats of above-mentioned each group after rat by taking anticoagulated whole blood is administered 90 days and 180 days, continue to collect blood, point
Every biochemical indicator is detected with automatic clinical chemistry analyzer from serum.24 hours after last time is administered, each group rat is living
10 rats retained after killing 10 stop administration, normal to raise, and continuing observation surrounding, all work is killed again.With medication 6 months
When the case where it is identical under conditions of, the index of detection is also identical.
After administration 90 days the result shows that: compared with blank control group, in, low dose group total protein (TP) content significantly rises
It is high, in which: low dose group increases obvious；Low dose group albumin content (ALB) significantly increases；High, medium and low dosage Histaglobin
(GIB) content significantly increases, in which: low dose group increases most obvious；High, middle dose group sodium ion (Na+) concentration significantly increases；
High dose potassium ion (K+) concentration significantly increases.Remaining index: as such as glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST), urine
Plain nitrogen (BUN), creatinine (CREA), blood glucose (GLU), total bilirubin (TBIL), total cholesterol (CHOL), triglycerides (TG), chlorine
Ion (Cl-) each administration group of concentration group difference compared with blank control group is unobvious, it does not make significant difference, the results are shown in Table 38.
Administration 180 days after the result shows that: compared with blank control group, in, low dosage aspartate amino transferase
(GOT), creatinine (CREA) content significantly reduces, in which: low dose group reduces most obvious；In, low dose group total protein (TP) contains
Amount is significant to be increased, in, low dose group albumin content (ALB) significantly increases；High and low dose group sodium ion (Na+), chloride ion
(Cl-) concentration significantly increases, in which: low dose group increases most obvious；Remaining index: such as alanine aminotransferase (ALT), alkali
Acid phosphatase (ALP), urea nitrogen (BUN), blood glucose (GLU), total bilirubin (TBIL), total cholesterol (CHOL), triglycerides
(TG) each administration group group difference compared with blank control group is unobvious, does not make significant difference, the results are shown in Table 39.
Surrounding convalescence the result shows that: compared with blank control group, middle dose group alkaline phosphatase (ALP) content significantly drops
It is low；High and low dose group total protein (TP) content significantly increases；High and low dose group albumin (ALB) content significantly increases；It is high and low
Dosage Histaglobin (GIB) content significantly increases, in which: low dose group increases most obvious；High dose group sodium ion (Na+) concentration
It is significant to increase；Remaining index: such as glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST), urea nitrogen (BUN), creatinine (CREA), flesh
Acid phosphoric acid kinases (CK), blood glucose (GLU), total bilirubin (TBIL), total cholesterol (CHOL), triglycerides (TG), potassium ion (K+), chloride ion (Cl-) each administration group of concentration group difference compared with blank control group is unobvious, it does not make significant difference, the results are shown in Table
40。
4) system postmortem and histopathologic examination's result
Administration 90 days after the result shows that: each administration group organ coefficient is substantially close with blank control group, illustrates drug to dirty
The amount of thinking highly of has no significant effect, and is shown in Table 41.The internal organs Histopathology microscopy of blank control group and each administration group rat, by sxemiquantitative
Scoring statistical result.The result shows that: except distilled water control group and medication each group minority example see have the slight heart, lung, renal interstitial,
Small enteremia and fatty degeneration of liver, and the congestive enlargement of a small number of example spleens, lymph node chronic inflammation, do not find related with drug toxicity
Lesion, and there are each internal organs of the animal of change to be dispersed in each group, be not statistically significant.Test result is shown in Table 42.
Administration 180 days after the result shows that: the organ coefficient of three dosage groups and blank control group are almost the same, explanation
Drug has no significant effect organ weights, is shown in Table 43.The internal organs Histopathology microscopy of blank control group and each administration group rat,
By sxemiquantitative scoring statistical result.The result shows that: except distilled water control group and medication each group minority example see have the slight heart,
Liver, bladder, lung and the congestive enlargement of a small number of example spleens, lymph node chronic inflammation do not find lesion related with drug toxicity, and have
Each internal organs of the animal of change are dispersed in each group, are not statistically significant.Experimental result is shown in Table 44.
Surrounding convalescence the result shows that: meticulously gross necropsy comprehensively, no abnormal organ-tissue are carried out to internal organs.Respectively
The group animal viscera heart, liver, spleen, lung, kidney, adrenal gland, uterus, brain, ovary, the organ coefficient of 10 organs and tissues such as thymus gland with it is right
It is substantially close according to organizing, organ weights are had no significant effect, are shown in Table 45.The internal organs pathologic group of blank control group and each administration group rat
Microscopy is knitted, by sxemiquantitative scoring statistical result.The result shows that: see have gently except distilled water control group and medication each group minority example
The heart of degree, small intestine, liver, kidney, lung and the congestive enlargement of a small number of example spleens, lymph node, stomach, large intestine chronic inflammation, do not find and drug
The related lesion of toxicity, and there are each internal organs of the animal of change to be dispersed in each group, it is not statistically significant.Experimental result is shown in Table 46.
Influence to rat blood biochemical indicator in 38 medication of table 3 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| ALT(U/L) | 80.30±53.66 | 61.50±18.82 | 57.70±16.26 | 67.00±18.97 |
| AST(U/L) | 218.20±143.70 | 148.20±36.96 | 151.60±23.32 | 174.00±38.28 |
| ALP(U/L) | 116.2±46.21 | 186.88±52.59 | 128.70±64.20 | 150.70±60.00 |
| CK(U/L) | 724.9±521.27 | 420.9±127.75 | 753.30±751.18 | 835.8±623.10 |
| TP(g/L) | 74.08±4.05 | 80.26±9.01 | 80.18±5.58* | 84.47±6.48*** |
| ALB(g/L) | 32.69±2.29 | 32.24±5.56 | 34.21±2.32 | 35.08±1.62* |
| GIB(g/L) | 41.39±2.74 | 48.02±8.90* | 45.97±4.93* | 49.39±5.66*** |
| BUN(mmol/L) | 6.37±1.22 | 7.78±2.42 | 7.26±1.44 | 6.55±1.66 |
| CREA(μmol/L) | 39.04±3.83 | 37.21±6.71 | 35.50±3.83 | 39.77±6.08 |
| GLU(mmol/L) | 5.12±0.69 | 4.41±0.99 | 4.68±0.86 | 5.05±0.61 |
| TG(mmol/L) | 0.56±0.22 | 0.87±0.44 | 0.74±0.86 | 0.64±0.27 |
| CHOL(mmol/L) | 1.78±0.32 | 1.63±0.30 | 1.93±0.35 | 1.86±0.27 |
| TBIL(μmol/L) | 0.60±0.37 | 0.62±0.40 | 0.66±0.37 | 0.45±0.24 |
| K+(mmol/L) | 4.51±0.48 | 5.84±0.52* | 5.02±0.92 | 5.47±1.37 |
| Na+(mmol/L) | 142.70±1.70 | 145.00±2.45* | 146.00±4.62* | 145.00±4.83 |
| Cl-(mmol/L) | 101.40±1.96 | 101.40±2.55 | 101.50±2.64 | 101.80±2.39 |
Note: compared with blank control group, * P < 0.05；* * P < 0.001.
Influence to rat blood biochemical indicator in 39 medication of table 6 months
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| GPT(U/L) | 111.6±86.89 | 100.9±42.01 | 83.4±35.06 | 72.90±13.39 |
| GOT(U/L) | 278.3±124.19 | 243.6±62.93 | 168.10±36.63 | 161.00±28.26** |
| AKP(U/L) | 214.3±105.37 | 197.8±92.24 | 238.40±112.70 | 265.40±109.97 |
| CK(U/L) | 2009.38±1301.08 | 1694.10±941.50 | 1449.30±712.6 | 1163.00±402.28 |
| BUN(mmol/L) | 6.53±1.17 | 5.80±1.42 | 5.73±0.34 | 5.54±1.13 |
| CR(μmol/L) | 28.93±10.01 | 36.02±23.18 | 15.97±6.11** | 18.85±8.38* |
| GLU(mmol/L) | 10.15±5.33 | 10.89±3.96 | 5.20±0.53 | 5.08±0.55 |
| TPR(g/L) | 67.2±4.07 | 63.20±4.10 | 81.29±6.83 | 77.97±4.22*** |
| ALB(g/L) | 31.84±2.53 | 31.80±10.54 | 36.13±3.51 | 35.38±1.98** |
| TBIL(μmol/L) | 0.82±0.58 | 0.55±0.91 | 1.22±0.67 | 1.13±0.32 |
| CHO(mmol/L) | 1.37±0.33 | 1.20±0.16 | 1.70±0.46 | 1.55±0.32 |
| TG(mmol/L) | 1.09±0.35 | 1.04±0.41 | 1.21±0.53 | 0.82±0.20 |
| K+(mmol/L) | 5.13±0.66 | 4.98±0.20 | 5.02±0.43 | 4.65±0.33 |
| Na+(mmol/L) | 137.92±1.70 | 140.15±2.11* | 139.20±1.93 | 141.70±3.68** |
| Cl-(mmol/L) | 99.77±1.41 | 100.32±2.57* | 99.10±1.79 | 101.50±3.10** |
Note: compared with blank control group, * P < 0.05；* P < 0.01；* * P < 0.001.
The variation of table 40 convalescence rat biochemical indicator
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| ALT(U/L) | 88.60±49.49 | 67.90±15.85 | 112.00±100.61 | 63.70±11.66 |
| AST(U/L) | 149.50±34.47 | 133.40±19.06 | 208.10±80.48 | 142.50±37.15 |
| ALP(U/L) | 211.70±82.95 | 184.50±35.35 | 128.7±29.28** | 248.70±99.95 |
| CK(U/L) | 635.30±196.11 | 873.90±292.15 | 838.20±305.99 | 926.30±729.52 |
| TP(g/L) | 70.70±3.68 | 76.22±2.64*** | 67.92±7.58 | 80.51±3.25 |
| ALB(g/L) | 31.401±1.68 | 33.94±2.05*** | 30.62±3.24 | 34.61±2.86* |
| GIB(g/L) | 38.77±3.02 | 42.28±2.74* | 37.30±5.38 | 45.90±3.73*** |
| BUN(mmol/L) | 7.67±1.39 | 7.65±0.95 | 6.79±1.44 | 7.28±0.96 |
| CREA(μmol/L) | 29.60±3.88 | 28.57±4.77 | 30.86±3.60 | 33.78±5.64 |
| GLU(mmol/L) | 6.21±1.19 | 5.03±0.38 | 6.46±1.57 | 5.34±0.77 |
| TG(mmol/L) | 0.70±0.28 | 0.99±0.42 | 0.74±0.41 | 1.02±0.46 |
| CHOL(mmol/L) | 1.37±0.32 | 1.45±0.22 | 1.47±0.32 | 1.68±0.35 |
| TBIL(μmol/L) | 0.23±0.13 | 0.27±0.19 | 0.26±0.36 | 0.32±0.23 |
| K+(mmol/L) | 5.72±0.95 | 6.10±0.56 | 5.44±0.73 | 6.44±0.52 |
| Na+(mmol/L) | 143.00±2.21 | 145.00±0.94* | 140.70±1.42 | 145.70±3.43 |
| Cl-(mmol/L) | 101.70±1.16 | 102.10±1.29 | 99.8±1.81 | 101.70±2.19 |
Note: compared with blank control group, * P < 0.05；* * P < 0.001.
41 medication of table 3 months to the influence to Rats Organs and Tissues coefficient
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| Heart percentage of liveweight percentage (%) | 0.37±0.06 | 0.35±0.07 | 0.35±0.09 | 0.37±0.05 |
| Liver percentage of liveweight percentage (%) | 3.23±0.61 | 2.93±0.57 | 3.34±0.74 | 3.04±0.42 |
| Spleen percentage of liveweight percentage (%) | 0.17±0.07 | 0.16±0.05 | 0.16±0.06 | 0.15±0.03 |
| Lung percentage of liveweight percentage (%) | 0.63±0.25 | 0.57±0.13 | 0.58±0.10 | 0.61±0.16 |
| Kidney percentage of liveweight percentage (%) | 0.30±0.04 | 0.28±0.08 | 0.27±0.05 | 0.28±0.05 |
| Adrenal gland percentage of liveweight percentage (%) | 0.0132±0.0050 | 0.0116±0.0050 | 0.01118±0.0031 | 0.0127±0.0027 |
| Thymus gland percentage of liveweight percentage (%) | 0.0707±0.0511 | 0.682±0.0324 | 0.0843±0.0202 | 0.0772±0.0276 |
| Brain percentage of liveweight percentage (%) | 0.6748±0.0915 | 0.6748±0.1308 | 0.6776±0.1235 | 0.6935±0.0976 |
| Testis percentage of liveweight percentage (%) | 0.48±0.10 | 0.41±0.11 | 0.41±0.13 | 0.45±0.03 |
| Epididymis percentage of liveweight percentage (%) | 0.23±0.08 | 0.25±0.05 | 0.23±0.04 | 0.23±0.04 |
| Prostate percentage of liveweight percentage (%) | 0.15±0.06 | 0.15±0.04 | 0.15±0.05 | 0.14±0.04 |
| Uterus percentage of liveweight percentage (%) | 0.30±0.072 | 0.31±0.106 | 0.31±0.120 | 0.28±0.150 |
| Ovary percentage of liveweight percentage (%) | 0.03±0.01 | 0.03±0.01 | 0.03±0.01 | 0.03±0.01 |
3 months rat pathological examination results are administered in table 42
Note: in table: "+" indicates that internal organs have Minimal change；"-" indicates that internal organs do not have apparent pathological change；Number (1~
10) number of elements of animal is indicated.
43 medication of table 6 months to the influence to Rats Organs and Tissues coefficient
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| Heart percentage of liveweight percentage (%) | 0.36±0.09 | 0.38±0.05 | 0.37±0.10 | 0.38±0.05 |
| Liver percentage of liveweight percentage (%) | 3.21±0.58 | 3.36±0.52 | 3.20±0.73 | 3.28±0.47 |
| Spleen percentage of liveweight percentage (%) | 0.17±0.06 | 0.15±0.03 | 0.15±0.04 | 0.18±0.07 |
| Lung percentage of liveweight percentage (%) | 0.50±0.07 | 0.48±0.08 | 0.47±0.09 | 0.54±0.08 |
| Kidney percentage of liveweight percentage (%) | 0.28±0.05 | 0.30±0.04 | 0.28±0.05 | 0.29±0.06 |
| Adrenal gland percentage of liveweight percentage (%) | 0.0121±0.0045 | 0.0125±0.0054 | 0.0108±0.0046 | 0.0120±0.0056 |
| Thymus gland percentage of liveweight percentage (%) | 0.0947±0.0330 | 0.0923±0.0317 | 0.0903±0.0272 | 0.1013±0.0412 |
| Brain percentage of liveweight percentage (%) | 0.5836±0.0913 | 0.5914±0.0488 | 0.5702±0.0791 | 0.5848±0.0985 |
| Testis percentage of liveweight percentage (%) | 0.49±0.08 | 0.51±0.10 | 0.47±0.05 | 0.53±0.09 |
| Epididymis percentage of liveweight percentage (%) | 0.16±0.03 | 0.16±0.05 | 0.14±0.02 | 0.17±0.05 |
| Prostate percentage of liveweight percentage (%) | 0.15±0.03 | 0.15±0.05 | 0.14±0.05 | 0.14±0.04 |
| Uterus percentage of liveweight percentage (%) | 0.20±0.06 | 0.18±0.07 | 0.20±0.02 | 0.24±0.10 |
| Ovary percentage of liveweight percentage (%) | 0.02±0.06 | 0.02±0.01 | 0.02±0.01 | 0.03±0.01 |
6 months rat pathological examination results are administered in table 44
Note: in table: "+" indicates that internal organs have Minimal change；"-" indicates that internal organs do not have apparent pathological change；Number (1~
10) number of elements of animal is indicated.
The variation of table 45 convalescence Rats Organs and Tissues weight
| Group | Control group | High dose group | Middle dose group | Low dose group |
| Number of animals (only) | 10 | 10 | 10 | 10 |
| Heart percentage of liveweight percentage (%) | 0.33±0.08 | 0.36±0.06 | 0.33±0.05 | 0.33±0.04 |
| Liver percentage of liveweight percentage (%) | 3.10±0.89 | 3.26±0.55 | 3.06±0.56 | 3.17±0.44 |
| Spleen percentage of liveweight percentage (%) | 0.17±0.05 | 0.20±0.06 | 0.17±0.06 | 0.19±0.07 |
| Lung percentage of liveweight percentage (%) | 0.54±0.12 | 0.53±0.08 | 0.54±0.09 | 0.54±0.09 |
| Kidney percentage of liveweight percentage (%) | 0.28±0.06 | 0.32±0.06 | 0.29±0.04 | 0.28±0.03 |
| Adrenal gland percentage of liveweight percentage (%) | 0.0101±0.0033 | 0.0099±0.0035 | 0.0100±0.0025 | 0.0104±0.0034 |
| Thymus gland percentage of liveweight percentage (%) | 0.0723±0.0322 | 0.0761±0.0155 | 0.0771±0.0216 | 0.0692±0.0416 |
| Brain percentage of liveweight percentage (%) | 0.5501±0.0549 | 0.5724±0.0470 | 0.5674±0.0520 | 0.5572±0.0719 |
| Testis percentage of liveweight percentage (%) | 0.4209±0.05 | 0.3938±0.07 | 0.3985±0.13 | 0.4290±0.07 |
| Epididymis percentage of liveweight percentage (%) | 0.1647±0.07 | 0.1429±0.06 | 0.1630±0.05 | 0.1529±0.05 |
| Prostate percentage of liveweight percentage (%) | 0.1456±0.05 | 0.1344±0.04 | 0.1607±0.05 | 0.1386±0.07 |
| Uterus percentage of liveweight percentage (%) | 0.3611±0.03 | 0.3909±0.06 | 0.3534±0.07 | 0.3371±0.08 |
| Ovary percentage of liveweight percentage (%) | 0.0212±0.008 | 0.02689±0.01 | 0.0241±0.01 | 0.0192±0.01 |
46 rat long term toxication convalescence of table puts to death animal pathological examination results
Note: in table: "+" indicates that internal organs have Minimal change；"-" indicates that internal organs do not have apparent pathological change；Number (1~
10) number of elements of animal is indicated.
The present composition with 10 times of quasi- dosage of clinic (0.45g/kg), 30 times of quasi- dosage of clinic (1.35g/kg),
60 times of quasi- dosage of clinic (2.7g/kg) give rat continuous gavage 180 days, and carry out experiment mid-term (administration 90 days) and 1 month
Convalescence Germicidal efficacy.The result is that:
(1) rat growthing development and general status are had not significant impact, only having at medicine feed initial stage a little influences, but
Restore after 2 weeks normal.
(2) to the influence of rat blood index
Successive administration 3 months, compared with blank control group, the present composition was to red blood cell, leucocyte in rat blood
Have a significant impact.Wherein: high dose group can be substantially reduced monocyte percentage in Rat Erythrocytes and leucocyte
(MONO%) content can significantly increase Rat Erythrocytes average external volume (MCV)；Low dose group significantly reduces drenches in rat leukocyte
The content of bar cell percentages (LYMPH%).
Successive administration 6 months, compared with blank control group, the present composition to red blood cell in rat blood, leucocyte,
Hemoglobin has a significant impact.Wherein: leucocyte in rat blood (WBC) content can obviously drop in high and low dose group；In, it is low
The content of monocyte percentage (MONO%) in the significant increasing leukocyte of dosage group energy；High, medium and low dosage group can significantly drop
Low red blood cell (RBC), hemoglobin (HGB) content and hematocrit (HCT)；
Be discontinued January after whole blood index restore normal.
(3) to the influence of rat blood biochemical indicator
Successive administration 3 months, compared with blank control group, the present composition was to albumen in rat blood and electrolyte
Content influences more apparent.Wherein: in, low dose group can significantly increase total protein in rat blood (TPR) content；Low dose group energy
Significant raising albumin content (ALB)；High, medium and low dosage group significantly increases globulin (GIB) content；High, middle dose group is shown
It writes and increases sodium ion (Na+) concentration and high dose group can also significantly increase potassium ion (K+) concentration.
Successive administration 6 months, compared with blank control group, the present composition was removed to albumen in rat blood and electrolyte
Content influence more apparent outer, be also affected to aspartate amino transferase (GOT) and creatinine (CREA) content.Its
In: in, low dose group can significantly increase total protein in rat blood (TP), albumin content (ALB) content；High and low dose group
Sodium ion (Na can significantly be increased+), chloride ion (Cl-) concentration.In, low dose group can significantly reduce Aspartate amino transfer
Enzyme (GOT), creatinine (CREA) content.
After being discontinued 1 month, rat blood Mid-Heaven Gate aspartic acid aminopherase (GOT), creatinine (CREA) restore normal；
Total protein (TPR), albumin (ALB) and globulin (GIB) content still significantly increase in high and low dose group；Middle dose group can be shown
Write the content for reducing alkaline phosphatase (ALP)；High dose group sodium ion (Na+) concentration still significantly increases.
(4) to Rats Organs and Tissues weight (heart, liver, spleen, lung, kidney, adrenal gland, thymus gland, uterus, ovary, testis, epididymis, forefront
Gland and brain totally 13 internal organs) it has no significant effect, each group organ coefficient is substantially close.To organs and tissues (in addition to above-mentioned 13 internal organs,
There are also pancreas, stomach, ileum, colon, pituitary, spinal cord, marrow, lymph node, bladder, sciatic nerve, thyroid gland, parathyroid gland,
Aorta, salivary gland, aorta) histopathologic examination, do not find that Pathomorphology significantly related with drug toxicity changes
Become.
Claims (9)
1. a kind of herbal mixture for treating myocardial ischemia and hyperlipidemia, it is characterised in that: the herbal mixture is by following parts by weight
Several Chinese medicine materials are made: 1~10 part of leaves of Hawthorn, 1~5 part of Longstamen Onion Bulb, 1~3 part of rhizoma polygonati.
2. herbal mixture according to claim 1, which is characterized in that the parts by weight of the Chinese medicine material are as follows: leaves of Hawthorn 7
Part, 3.5 parts of Longstamen Onion Bulb, 1.5 parts of rhizoma polygonati.
3. herbal mixture according to claim 1 or 2, it is characterised in that: the dosage form that the herbal mixture is prepared into is to appoint
What is a kind of clinically or pharmaceutically acceptable dosage form.
4. herbal mixture according to claim 3, it is characterised in that: the dosage form is dripping pill, tablet, capsule, particle
Agent, oral liquid or injection.
5. according to claim 1 to 2 described in any item herbal mixtures, it is characterised in that the following steps are included:
Each with water or alcohol to leaves of Hawthorn, Longstamen Onion Bulb and rhizoma polygonati individually carries out one or many effective component and extracts；
Or one or many effective component is carried out to the mixture of leaves of Hawthorn, Longstamen Onion Bulb and rhizoma polygonati with water or alcohol and is extracted.
6. according to claim 1 to the preparation method of 2 described in any item herbal mixtures, it is characterised in that:
(1) amount weighs leaves of Hawthorn according to the ratio, after ethyl alcohol soaking time 0.1~10 hour of 2 times~40 times of leaves of Hawthorn weight, In
It is extracted 0.1~10 hour under conditions of 30 DEG C~100 DEG C, the ethyl alcohol impregnates and extraction operation can carry out 1 time or repeat
Up to 6 times, combined extract is distilled to recover solvent, and centrifugation is washed with the purifying of 0.1 times~100 times volumes of leaves of Hawthorn medicinal material amount
It washs precipitating once or washes repeatedly up to five times, merge the supernatant after being centrifuged and wash the supernatant precipitated, distillation and concentration is extremely
After close dry, dry, pulverize to get；
(2) amount weighs Longstamen Onion Bulb and rhizoma polygonati according to the ratio, and Longstamen Onion Bulb and rhizoma polygonati total weight 2 are added for the first time under the conditions of 50 DEG C~100 DEG C
Times~40 times of water extract 0.1~10 hour, extraction process can carry out once or up to 5 times, merging and extracting gained decocting liquid, filtering is simultaneously
Concentrate the filtrate to Longstamen Onion Bulb and 0.1 times~20 times of rhizoma polygonati total weight, ethyl alcohol added to make alcohol content 20%~90%, stand or from
The heart takes supernatant, filtering, be distilled to recover ethyl alcohol, continue distillation and concentration at thick paste, dry, pulverize to get；
(3) Longstamen Onion Bulb, the Rhizoma Polygonati extract that the hawthorne leaf P.E and above-mentioned steps (2) obtained according to above-mentioned steps (1) obtains are mixed
After closing uniformly, additives appropriate are added, injection, oral solid formulation or oral liquid, the oral administration solid is made
Preparation is tablet, capsule, granule, and the oral liquid is oral solution or syrup, and the additives are filling
One of agent, excipient, corrigent or preservative are a variety of.
7. the preparation method of herbal mixture according to claim 6, it is characterised in that:
(1) 7 parts of leaves of Hawthorn are weighed, for the first time with 8 times of 45% ethyl alcohol soaking time of leaves of Hawthorn weight after 2 hours, at 80 DEG C
Under conditions of extract 2 hours, second is extracted 1 hour with 5 times of 45% ethyl alcohol of leaves of Hawthorn weight in 80 DEG C of condition, third
It is secondary with 4 times of leaves of Hawthorn weight measure 45% ethyl alcohol 80 DEG C condition extract 0.5 hour, the 4th time with 3 times of leaves of Hawthorn weight measure
45% ethyl alcohol extracts 0.5 hour in 80 DEG C of condition, and combined extract is distilled to recover ethyl alcohol, and centrifuge is 3000R's per minute
It is centrifuged 30min under revolving speed, is precipitated three times with the purifying water washing of 2 times of volumes of medicinal material amount, supernatant and washing after merging centrifugation
The supernatant of precipitating, distillation and concentration to dry, pulverize after close dry to get；
(2) 3.5 parts and 1.5 parts of rhizoma polygonati of Longstamen Onion Bulb are weighed, 10 times of Longstamen Onion Bulb and rhizoma polygonati total weight are added for the first time under the conditions of 100 DEG C
Water decocts 2 hours, and 8 times of water of second of addition Longstamen Onion Bulb and rhizoma polygonati total weight decoct 1 hour, and collecting decoction, filtrate is concentrated into medicinal material
1 times of total weight, ethyl alcohol is added to make alcohol content 55%, stands or be centrifuged, take supernatant, filter, be distilled to recover ethyl alcohol, continue to distill
Be condensed into thick paste, dry, pulverize to get；
(3) after mixing by the hawthorne leaf P.E of step (1) acquisition and the Longstamen Onion Bulb of step (2) acquisition, Rhizoma Polygonati extract, add
Enter additives appropriate, 10 portions of injections, oral solid formulation or oral liquid, the additives are made are as follows: starch,
Lactose, sodium carboxymethylcellulose, magnesium stearate, one of sucrose, stevioside, sodium benzoate, soybean oil or soybean lecithin or
It is a variety of.
8. the preparation method of herbal mixture according to claim 7, it is characterised in that:
(1) by the hawthorne leaf P.E of claim 7 step (1) acquisition, after the petroleum ether degreasing with 1/2 amount, petroleum ether is discarded
Liquid, then be extracted with ethyl acetate, extract liquor is recovered under reduced pressure ethyl acetate and is concentrated to dryness, and suitable quantity of water, which is added, to be made to dissolve, and is added on poly-
On amide column, first with the water elution of 2 times of column volumes, water lotion is discarded, then with 80% ethanol elution of 3 times of column volumes, is collected
Eluent recycles ethyl alcohol and is concentrated into the concentrate of 60 DEG C of relative densities about 1.02~1.08, dry, obtains haw thorn leaf total flavone and mentions
Take object；
(2) Longstamen Onion Bulb, the Rhizoma Polygonati extract obtained claim 7 step (2), 95% ethanol precipitation accumulated with diploid, instead
It carries out 4 times again, falls small-molecule substance with dialysis membrane dialysis, alcohol precipitation again, finally with the anhydrous ether of equal volume and suitable
Acetone washing sediment, it is dry, obtain Longstamen Onion Bulb, coarse solomon's seal polysaccharide extract；
(3) Longstamen Onion Bulb, the rhizoma polygonati that the haw thorn leaf total flavone extract and above-mentioned steps (2) obtained above-mentioned steps (1) obtains are slightly more
After mixing, additives appropriate are added in sugar extract, and 10 parts of powder-injection, small-volume injection or large capacity injection is made
Agent.
9. the preparation method of herbal mixture according to claim 8, which is characterized in that when injection is made, in order to increase
Add its solubility, Tween-80 solubilizer is added；Excipient is added in powder needle, the excipient is mannitol or glucose；Infusion
The isotonic regulator for adjusting osmotic pressure is added in preparation, the isotonic regulator is sodium chloride, potassium chloride, magnesium chloride, chlorine
Change one of calcium, sodium lactate, glucose, xylitol, sorbierite and dextran or a variety of.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310006120.8A CN103908571B (en) | 2013-01-08 | 2013-01-08 | A kind of Chinese traditional compound medicine for treating heart disease |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310006120.8A CN103908571B (en) | 2013-01-08 | 2013-01-08 | A kind of Chinese traditional compound medicine for treating heart disease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103908571A CN103908571A (en) | 2014-07-09 |
| CN103908571B true CN103908571B (en) | 2019-11-05 |
Family
ID=51034775
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310006120.8A Active CN103908571B (en) | 2013-01-08 | 2013-01-08 | A kind of Chinese traditional compound medicine for treating heart disease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103908571B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104510903A (en) * | 2013-10-08 | 2015-04-15 | 肖延斌 | Compound traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases |
| CN109115543B (en) * | 2018-07-09 | 2021-06-22 | 苏州华测生物技术有限公司 | Method for effectively improving pathological slicing cut-out rate of parathyroid gland tissue of experimental animal |
| CN109323910B (en) * | 2018-12-03 | 2021-01-08 | 徐州市中心医院 | Preparation method of high-purity low-density lipoprotein |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048217A (en) * | 2009-10-30 | 2011-05-11 | 常堡同 | Beverage capable of reducing blood pressure and blood fat and production method thereof |
| CN102100833A (en) * | 2011-01-24 | 2011-06-22 | 四川国康药业有限公司 | Drug composition for treating heart cerebrovascular diseases as well as preparation method and application thereof |
-
2013
- 2013-01-08 CN CN201310006120.8A patent/CN103908571B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102048217A (en) * | 2009-10-30 | 2011-05-11 | 常堡同 | Beverage capable of reducing blood pressure and blood fat and production method thereof |
| CN102100833A (en) * | 2011-01-24 | 2011-06-22 | 四川国康药业有限公司 | Drug composition for treating heart cerebrovascular diseases as well as preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103908571A (en) | 2014-07-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101091765A (en) | Medicinal comsns-and usage for preventing and treating diabets mellitus | |
| CN1931217B (en) | Medicine composition of gingko leaf and rhodiola root | |
| CN107441078A (en) | A kind of pharmaceutical composition for treating diabetes and its production and use | |
| CN1327876C (en) | Chinese drugs compositions for treating diabetes and its complications | |
| CN104435034B (en) | A kind of arasaponin and preparation method thereof | |
| CN101390887A (en) | Medicine composition of Panax notoginseng saponins | |
| CN103919854B (en) | Application of butterflybush flower and extract thereof to preparation of medicament | |
| CN103908571B (en) | A kind of Chinese traditional compound medicine for treating heart disease | |
| CN101152285B (en) | Pharmaceutical composition of snakegourd fruit and whitethorn leaf | |
| CN1931233B (en) | Medicine composition of red sage and epimedium for treating cardiac and cerebral vascular diseases | |
| CN104510884A (en) | Compound traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases | |
| CN102475766B (en) | Drug composition for treating heart failure, preparation method and application of drug composition | |
| CN1935178A (en) | Ginkgo leaf extract and barrenwort extract composition | |
| CN1931216B (en) | Medicine composition of safflower and rhodiola root | |
| CN1939383B (en) | Use of redback christmashush in preparing medicine for hepatitis B | |
| CN101152233A (en) | Pharmaceutical composition of snakegourd fruit and folium ginkgo | |
| CN100563682C (en) | A kind of pharmaceutical composition of making by Folium Crataegi and Radix Rhodiolae and preparation method thereof | |
| CN106727498A (en) | Treat the composition and its preparation and the method for inspection of cardiovascular and cerebrovascular disease | |
| CN104510901A (en) | Compound traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases | |
| CN104510902A (en) | Compound traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases | |
| CN1081465C (en) | Rhodiola rosea injection for treating angina pectoris of coronary heart disease | |
| CN104127545B (en) | New application of murraya tetramera huang and extract thereof in preparation of medicines | |
| CN106619972A (en) | Hirsutella hepiali chen et shen mycelium powder hypoglycemic capsule and preparation method and application thereof | |
| CN104510903A (en) | Compound traditional Chinese medicine preparation for treating cardiovascular and cerebrovascular diseases | |
| CN102091131B (en) | New application of lotus plumule or extract thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| DD01 | Delivery of document by public notice |
Addressee: Xiao Yanbin Document name: Notification of Passing Examination on Formalities |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |