CN104586931A - Traditional Chinese medicine for treatment of myocardial ischemia and preparation method as well as quality test method thereof - Google Patents
Traditional Chinese medicine for treatment of myocardial ischemia and preparation method as well as quality test method thereof Download PDFInfo
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Abstract
The invention belongs to the field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine composition for treatment of myocardial ischemia. The title of invention is a traditional Chinese medicine for treatment of myocardial ischemia and preparation method as well as a quality test method thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: adding Twoanther Mosla in a volatile oil extractor for distillation, extracting volatile oil, and obtaining the Twoanther Mosla volatile oil for later use; collecting the aqueous solution formed after distillation of the Twoanther Mosla in another container for later use; adding water for decocting the Twoanther Mosla dregs formed after volatile oil extraction with Lysidice rhodostegia Hance, Schoepfia jasminodora sieb.et and beet root for 2-4 times, decocting for 0.5-2 hours each time, combining decoction solution, filtering, combining filtrate with the aqueous solution formed after distillation of the Twoanther Mosla, performing static settlement for 12-36 hours, filtering, concentrating filtrate to form dilute paste with relative density of 1.00-1.10 at 60 degrees centigrade, adding sucrose powder and dextrin into the dilute paste, placing the mixture into a dry granulator to prepare granules, adding the Twoanther Mosla volatile oil, and mixing evenly, thereby obtaining the traditional Chinese medicine composition. The traditional Chinese medicine composition can be used for preparing the traditional Chinese medicine for treatment of myocardial ischemia.
Description
technical field:
The invention belongs to the field of Chinese medicines, relate to a kind of Chinese medicine composition with treatment myocardial ischemia be made up of miliaria alba grass, ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae.
technical background:
Myocardial ischemia refers to that the hemoperfusion of heart reduces, and cause the oxygen supply of heart to reduce, energy metabolism of myocardial is abnormal, can not support a kind of pathological state of normal heart action.Coronary heart disease causes main, the modal cause of disease of myocardial ischemia.Along with the raising of living standards of the people, current myocardial ischemia is the trend risen year by year in the prevalence of China.Myocardial ischemia is commonly encountered diseases and the frequently-occurring disease of middle-aged and elderly people.
The common reason of myocardial ischemia is coronary atherosclerosis, secondly also has inflammation (rheumatic, syphilis, mucocutaneous lymphnode syndrome and thromboangitis obliterans etc.), spasm, thromboembolism, connective tissue disease, wound and congenital malformation etc. multiple.Epidemiological study finds, the important risk factor relevant to atherosclerosis is hyperlipemia, hypertension, diabetes, smoking, obesity, homocysteine increases, physical exertion is few, advanced age and male etc.
Clinical manifestation: 1. after there is breastbone during tired or psychentonia or precordial fullness pain, or tighten sample pain, and take on left, left upper arm radiation, continue 3 ~ 5 minutes, spontaneous remission person after having a rest, time with profuse sweating.2. there is uncomfortable in chest, cardiopalmus during physical exertion, breathe hard, spontaneous remission person during rest.3. occur and the relevant throat pain of motion and burn feeling, constriction, have a toothache.4. heavy meal, cold, drink after there is chest pain, person uncomfortable in chest.5. when sleeping at night medicated pillow is low, feel chest distress, need high pillow clinostatism side to feel comfortable person; Sleep soundly or daytime is flat sleeping time unexpected chest pain, cardiopalmus, dyspnea, need sit up immediately or stand can the person of alleviation.6. sexual life or firmly defecation time occur nervous, uncomfortable in chest, out of breath or chest pain is uncomfortable.7. the bradycardia happened suddenly, blood pressure reduce or the person of fainting.
summary of the invention:
The object of this invention is to provide a kind of Chinese medicine composition for the treatment of myocardial ischemia.
Another object of the present invention is to provide the preparation method of Chinese medicine composition.
The present invention also provides the quality determining method of this Chinese medicine composition.
The present invention also provides the application of this Chinese medicine composition in preparation treatment myocardial ischemia and myocarditis Chinese medicine.
The object of the invention is to realize in the following manner:
Treating a Chinese medicine composition for myocardial ischemia, is be made up of the raw material of following weight portion: miliaria alba grass 1 ~ 5, ferrum Brassaiopsis glomerulata (Bl.) Regel 1 ~ 5, gristle wind 1 ~ 5, Radix Betae 1 ~ 5.
Described Chinese medicine composition is preferably made up of the raw material of following weight portion: miliaria alba grass 2, ferrum Brassaiopsis glomerulata (Bl.) Regel 3, gristle wind 2, Radix Betae 3.
Described Chinese medicine composition is preferably made up of the raw material of following weight portion: miliaria alba grass 3, ferrum Brassaiopsis glomerulata (Bl.) Regel 2, gristle wind 3, Radix Betae 2.
The miliaria alba grass 1 that described Chinese medicine composition is preferably made up of the raw material of following weight portion, ferrum Brassaiopsis glomerulata (Bl.) Regel 1, gristle wind 1, Radix Betae 1.
Described Chinese medicine composition adopts pharmaceutical methods conventional in pharmacy to be prepared into oral formulations.
The oral formulations that described Chinese medicine composition is prepared into is tablet, pill, hard capsule, granule, oral liquid.
Described Chinese medicine composition is adopted and is prepared with the following method: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 2 ~ 4 times, each 0.5 ~ 2h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 12 ~ 36h; filter; it is the clear paste of 1.00 ~ 1.10 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin, puts in spray-drying pelleting machine; make granule; add miliaria alba grass volatile oil, mixing, to obtain final product.
Described Chinese medicine composition is preferably adopted and is prepared with the following method: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin, puts in spray-drying pelleting machine; make granule; add miliaria alba grass volatile oil, mixing, to obtain final product.
The detection method of described Chinese medicine composition is:
(1) chromatographic condition: adopt C18 chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg.
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
Described Chinese medicine composition can be used for the Chinese medicine preparing treatment myocardial ischemia.
Described Chinese medicine composition can be used for the myocarditic Chinese medicine of preparation treatment.
Technique effect of the present invention is verified by following experimentation:
Experimental example one: the pharmacodynamic experiment research of Traditional Chinese Medicine Anti myocardial ischemia of the present invention
1 material
1.1 animal
Kunming mouse, male and female half and half, body weight 19 ~ 22g; Wistar rat, male and female half and half, body weight 200 ~ 220g, provides by University Of Yantai's Experimental Animal Center.
1.2 medicines and reagent
Chinese medicine of the present invention: get dry miliaria alba grass 250.0g, ferrum Brassaiopsis glomerulata (Bl.) Regel 250.0g, gristle wind 250.0g ， Radix Betae 250.0g, heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter; aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h, filters; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities; qinghuo reagent is put in spray-drying pelleting machine, makes granule, adds miliaria alba grass volatile oil; mixing, to obtain final product.Put Refrigerator store, be mixed with desired concn with distilled water before use.
Contrast Chinese medicine I: get dry miliaria alba grass 333.3g, ferrum Brassaiopsis glomerulata (Bl.) Regel 333.3g, gristle wind 333.3g, heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle wind decoct with water 3 times, each 1h, collecting decoction; filter; aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h, filters; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities; qinghuo reagent is put in spray-drying pelleting machine, makes granule, adds miliaria alba grass volatile oil; mixing, to obtain final product.Put Refrigerator store, be mixed with desired concn with distilled water before use.
Contrast Chinese medicine II: get dry ferrum Brassaiopsis glomerulata (Bl.) Regel 333.3g; gristle wind 333.3g Radix Betae 333.3g, gets ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decocts with water 3 times, each 1h; collecting decoction; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent is put in spray-drying pelleting machine; make granule, to obtain final product.Put Refrigerator store, be mixed with desired concn with distilled water before use.
Contrast Chinese medicine III: get dry gristle wind 333.3g ， Radix Betae 333.3g, miliaria alba grass 333.3g, heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter; aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h, filters; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities; qinghuo reagent is put in spray-drying pelleting machine, makes granule, adds miliaria alba grass volatile oil; mixing, to obtain final product.Put Refrigerator store, be mixed with desired concn with distilled water before use.
Contrast Chinese medicine IV: get dry Radix Betae 333.3g, miliaria alba grass 333.3g, ferrum Brassaiopsis glomerulata (Bl.) Regel 333.3g, heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter; aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h, filters; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities; qinghuo reagent is put in spray-drying pelleting machine, makes granule, adds miliaria alba grass volatile oil; mixing, to obtain final product.Put Refrigerator store, be mixed with desired concn with distilled water before use.
FUFANG DANSHEN PIAN (Beijing Science and Technology Ltd. of Tongrentang).
Glutamic oxaloacetic transaminase, GOT (AST), lactic acid dehydrogenase (LDH), creatine kinase isozyme (CK-MB) test kit (Bioengineering Research Institute is built up in Nanjing).
1.3 instrument
ECG6511 type electrocardiograph (Shanghai Photoelectricity Medical electron Instrument Co., Ltd.), BP-211D type electronic balance (Sartorius AG), 80-2 type table-type low-speed centrifuge (Shanghai Medical apparatus company limited), SABA-18 type automatic clinical chemistry analyzer (Italian AMS company).
2 methods
2.1 rat coronary artery ligation causes Acute Myocardial Ischemia in Rats
2.1.1 divide into groups, administration and modeling: adopts literature method (Li Jufeng, Lin Shaozhu, Tang Shaoli, etc. Rhizoma Zingiberis Recens saponin injection function of resisting myocardial ischemia studies ［ J ］. Chinese crude drug .2013.36(8): 1322) set up Model Rats with Acute Myocardial Ischemia.Get rat 80, be divided into 8 groups at random, sham operated rats (normal saline 20mgkg
-1), model group (normal saline 20mgkg
-1), Chinese drug-treated group (220mgkg of the present invention
-1), contrast Chinese medicine I group (220mgkg
-1), contrast Chinese medicine II group (220mgkg
-1), contrast Chinese medicine III group (220mgkg
-1), contrast Chinese medicine IV group (220mgkg
-1) and FUFANG DANSHEN PIAN group (250mgkg
-1), often organize 10.After Rat Fast 12h, ig administration, administration every day 1 time, continuous 7d.1h after last administration, with 12% chloral hydrate (360mgkg
-1) ip anesthesia, after measuring normal II lead electrocardiogram, cut skin, muscle layer, circular layer makes purse string suture, cut off the 4th rib, heart is extruded thoracic cavity, between pulmonary conus and left auricle, find out left anterior descending coronary artery, except sham operated rats only threading not ligation, all the other are respectively organized and all cause myocardial ischemia with No. 5/0 line ligation immediately.Sew up wound after ligation, with syringe pump breast intracavity gas before sewing up completely, animal is steam again and raises.More than perform the operation and all aseptically operate.
2.1.2 ECG ST section change: to record respectively before ligation and after ligation 5, II lead electrocardiogram of 30,60,120min, with preoperative II lead electrocardiogram as normal ECG, with the changing value of Rat Ecg ST section T ripple after administration for representing degree of myocardial ischemia.
2.1.3 the detection of Enzyme target in serum: get blood in rat aorta after 24h, centrifugal, separation of serum, illustrates mensuration serum AST by test kit, LDH, CK-MB level.
2.1.4 myocardial infarct size measures: put to death rat and win heart, normal saline flushing, weigh quality whole-heartedly.Become the left ventricular tissues of 1 ~ 2mm to cut into slices from the apex of the heart to heart base portion crosscut, normal saline is cleaned, and is placed in 1%TTC(triphenyltetrazolium chloride) solution, 37 DEG C of water-bath dyeing 15min.Normal saline flushing excess dyestuff, do not dye in infarcted region, non-infarcted region is dyed to redness, uncoloured infarcted region cut, and claims weight in wet base, calculates left ventricle infarction size (IS).
IS=infarcted myocardium quality/left ventricular mass × 100%
The airtight hypoxia endurance test of 2.2 mice
Mice 70, is divided into 7 groups at random, is respectively normal saline group, Chinese drug-treated group (220mgkg of the present invention
-1), contrast Chinese medicine I group (220mgkg
-1), contrast Chinese medicine II group (220mgkg
-1), contrast Chinese medicine III group (220mgkg
-1), contrast Chinese medicine IV group (220mgkg
-1) and FUFANG DANSHEN PIAN group (360mgkg
-1).Mice fasting 12h, ig administration, administration every day 1 time, continuous 7d.1h after last administration, puts into the airtight wide mouthed bottle of 250mL volume respectively, puts into sodica calx 10g in bottle by mice, bottleneck is coated with vaseline.From putting into mice, bottle covered and enclosed is started timing, the time-to-live of record mice.
2.3 statistical analysis
Adopt SPSS13.0 software analysis, the data obtained represents with ± s, compares and adopt t inspection, with P < 0.05 for there being statistical significance between group.
3 results
3.1 impacts on ECG ST section
As shown in table 1, compare with sham operated rats, after model group rats coronary ligation, ECG ST section occurs that significance raises > 0.1mV(P < 0.01 at once).Compare with model group, Chinese drug-treated group of the present invention can resist raising of ECG ST section caused by coronary ligation, the amplitude of raising (P < 0.05) of 30 ~ 120min ECG ST section after remarkable reduction ligation, antagonism effect is even better than FUFANG DANSHEN PIAN.Contrast Chinese medicine I group then not obvious to contrast Chinese medicine IV group action effect.
Table 1 Chinese medicine of the present invention on the impact of rats with myocardial ischemia ECG ST section (
± s, n=10)
Note: compare * P < 0.05, * * P < 0.01 with sham operated rats; #P < 0.05, ##P < 0.01 is compared with model group
3.2 impacts on Serum fibrosis markers
As shown in table 2, compare with sham operated rats, model group rats serum AST, LDH, CK-MB vigor all significantly raises (P < 0.01).Chinese drug-treated group of the present invention can reduce the rising of the serum cardiac enzyme activity caused by coronary ligation, significantly can reduce the rising (P < 0.01) of the serum cardiac enzyme activity caused by coronary ligation.Contrast Chinese medicine I group then not obvious to contrast Chinese medicine IV group action effect.
Table 2 Chinese medicine of the present invention on the impact of rats with myocardial ischemia Serum fibrosis markers (
± s, n=10)
Note: compare * P < 0.05, * * P < 0.01 with sham operated rats; #P < 0.05, ##P < 0.01 is compared with model group
3.3 impacts on myocardial infarct size
As shown in table 3, compare with sham operated rats, model group rats myocardial infarct size significantly raises (P < 0.01); Compare with model group, Chinese drug-treated group myocardial infarct size of the present invention significantly reduces (P < 0.01).Contrast Chinese medicine I group then not obvious to contrast Chinese medicine IV group action effect.
Table 3 Chinese medicine of the present invention on the impact of rats with myocardial ischemia myocardial infarct size (
± s, n=10)
| Group | Dosage/mgkg -1 | Left ventricle weight/mg | Infarcted region weight/mg | Infarction size/% |
| Sham operated rats | － | 960.95±126.25 | 00.00±00.00 | 00.00±00.00 |
| Model group | － | 974.33±121.54 | 163.31±70.96 | 16.76±8.31** |
| Chinese drug-treated group of the present invention | 200 | 1004.23±141.73 | 102.34±93.15 | 10.16±6.77## |
| Contrast Chinese medicine I group | 200 | 1007.35±123.16 | 142.78±61.00 | 14.19±4.58 |
| Contrast Chinese medicine II group | 200 | 1005.24±136.53 | 140.42±82.11 | 13.92±9.63 |
| Contrast Chinese medicine III group | 200 | 1009.39±123.18 | 147.57±61.06 | 14.65±4.54 |
| Contrast Chinese medicine IV group | 200 | 1005.20±136.59 | 140.47±82.16 | 13.95±9.64 |
| FUFANG DANSHEN PIAN group | 250 | 1009.44±140.73 | 103.92±79.41 | 10.32±5.63## |
Note: compare * P < 0.05, * * P < 0.01 with sham operated rats; #P < 0.05, ##P < 0.01 is compared with model group
3.4 impacts on mice hypoxia-bearing capability
As shown in table 4, the time-to-live of positive drug FUFANG DANSHEN PIAN energy significant prolongation mice, demonstrate the feasibility of this experimental system.After successive administration 7d, Chinese drug-treated group of the present invention has the effect (P < 0.05) of the time-to-live obviously extending mice, suitable with FUFANG DANSHEN PIAN effect.Contrast Chinese medicine I group then not obvious to contrast Chinese medicine IV group action effect.
Table 4 Chinese medicine of the present invention on the impact of mice oxygen deficit tolerance time-to-live (
± s, n=10)
| Group | Dosage/mgkg -1 | Time-to-live/s |
| Normal saline group | － | 906±126 |
| Chinese drug-treated group of the present invention | 200 | 1078±137* |
| Contrast Chinese medicine I group | 200 | 919±100 |
| Contrast Chinese medicine II group | 200 | 987±99 |
| Contrast Chinese medicine III group | 200 | 916±105 |
| Contrast Chinese medicine IV group | 200 | 984±93 |
| FUFANG DANSHEN PIAN group | 360 | 1022±112* |
Note: compare * P < 0.05, * * P < 0.01 with normal saline group.
4 conclusions
In sum, Chinese medicine of the present invention all has comparatively positive effect to Acute Myocardial Ischemia in Rats and the airtight anoxia enduring of mice, shows that Chinese medicine of the present invention has certain protective effect to ischemic myocardium.And be better than contrast Chinese medicine I, contrast Chinese medicine II, contrast Chinese medicine III, contrast Chinese medicine IV, illustrate that the compatibility in pharmaceutical composition of the present invention between each flavour of a drug is superior, indispensable, combination between each flavour of a drug creates obvious synergistic function, has the prospect being developed to the new Chinese medicine that resists myocardial ischemia.
Experiment two: treatment by Chinese herbs silent myocardial ischemia 30 example of the present invention
Silent myocardial ischemia (SMI) is very common in patients with coronary heart disease, also known as silent myocardial ischemia, refer to that coronary artery pathological changes or coronary vasospasm patient have the objective evidence of myocardial ischemia, but lack the coronary heart disease of angina pectoris clinical symptoms, it is a kind of specific type of coronary heart disease, because it is invisible easily out in the cold, it may develop into the serious cardiac event such as acute myocardial infarction, malignant arrhythmia, heart failure, cardiogenic shock, sudden death, and its potential danger is very big.
1 data and method
1.1 physical data
Choose SMI patient 60 example in year October my institute in January, 2014 to 2014, there is clear and definite history of coronary heart disease, all meet SMI diagnostic criteria (as if Chen Xin. yellow clinical electrocardiography [M]. Beijing: People's Health Publisher .2009:75-79): ST section is that horizontal type or declivity type move down >=0.1mv, persistent period >=1min, except other heart diseases; All cases are all difficult lower situation and occur: acute myocardial infarction in nearly month, nearly classical angina outbreak in February, heart failure, hepatic and kidney function obstacle, severe arrhythmia; The inspections such as all patient's row 24 hr Ambulatory EKG Monitoring, routine blood test, blood fat, blood glucose, hepatic and renal function, Color Sonography.Above patient is divided into treatment group and matched group at random, often organize each 30 example, wherein treatment group man 16 example, female 14 example, 47 ~ 69 years old age, the mean age (55 ± 6) year, the course of disease 1 ~ 5 year; Matched group man 15 example, female 15 example, 45 ~ 69 years old age, the mean age (54 ± 8) year, the course of disease 1 ~ 5 year, two groups of physical data no significant difference (P > 0.05).
1.2 method
60 routine SMI patients give the conventional therapies such as nitric acid lipid, aspirin, beta-blocker, treatment group adds with Chinese medicine 0.6Bid of the present invention on basis of conventional therapy, serve on six weeks, before and after paired observation treatment, two groups of blood fat, c reactive protein (CRP), ambulatory electrocardiograms change.
1.3 criterion of therapeutical effect
Effective: transference cure or substantially disappear, electrocardiogram recovers normally or roughly normal;
Effective: symptom obviously alleviates, electrocardio diagram ST section forces down rise >=0.05mv, and dynamic electrocardiogram diagram myocardial ischemic attacks number of times reduces over half;
Invalid: symptom is improved not obvious, electrocardiogram improvement does not reach effective standard.
1.4 statistical procedures
Application SPSS17.0 software processes, measurement data is used (
± s) represent, adopt t inspection, numeration data χ
2inspection, P < 0.05 has statistical significance for difference.
2 results
Experimental result is in table 5 and table 6.
Table 5 liang group treatment front and back blood fat and CRP change are compared (
± s)
Note: compare with before treatment, * P < 0.05, compares with matched group, #P < 0.05.
Table 6 liang group comparitive study (n, %)
| Group | Number of cases | Effective | Effectively | Invalid | Total effective rate |
| Treatment group | 30 | 15 | 13 | 2 | 93.3 |
| Contrast groups | 30 | 9 | 11 | 10 | 66.7 |
P < 0.01, treatment group total effective rate is significantly higher than matched group
3 conclusions
This research SMI gives treatment by Chinese herbs of the present invention on coronary dilating, antiplatelet aggregation basis, TC, LDL-L, TG, CRP level, the clinical symptoms improvement of result display treatment group are all obviously better than matched group (P < 0.05), treatment group total effective rate 93.3%, matched group total effective rate 66.7%, treatment group total effective rate is significantly higher than matched group, observed result is without liver function injury, without untoward reaction such as creatine phosphokinase rising and myalgias, treatment by Chinese herbs silent myocardial ischemia clinical efficacy of the present invention is definite, drug safety, clinic is promoted.
Experiment three: the preparation of Chinese medicine granules of the present invention and quality determining method are studied
1 instrument and reagent
1.1 instrument
High performance liquid chromatograph (Agilent110 high performance liquid chromatograph and work station, G1311Aquat pump, G1314 UV-detector); PGL-5A type spray-drying pelleting machine (Chongqing Seiko pharmaceutical machine Co., Ltd).
1.2 reagent
Absinthol reference substance (Chinese pharmaceutical biological product calibrating academy); Chinese medicine granules of the present invention: self-control; Methanol (chromatograph alcohol, biochemical work auxiliary reagent factory, Shanghai); Other reagent are analytical pure.
2 methods and result
2.1 prescription
Miliaria alba grass 250.0g, ferrum Brassaiopsis glomerulata (Bl.) Regel 250.0g, gristle wind 250.0g ， Radix Betae 250.0g
2.2 preparation
Heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin; put in spray-drying pelleting machine, make granule, add miliaria alba grass volatile oil; mixing, makes 1000g, is sub-packed in plastic bag; every bag of 5g, to obtain final product.
The assay of 2.3 absinthols
2.3.1HPLC chromatographic condition
Adopt C18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: methanol-acetonitrile (40:60); Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L.Under this chromatographic condition, reference substance and sample chromatogram peak are well, noiseless to mensuration without miliaria alba grass negative control.
2.3.2 the preparation of reference substance solution
It is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and make the solution of every 1mL containing 0.2mg.
2.3.3 the preparation of need testing solution and negative controls
Precision takes Chinese medicine granules 10g of the present invention, adds methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merges n-butanol extracting liquid, washs 3 times with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product.Separately do not contain the granule agent negative controls of miliaria alba grass in the preparation of prescription ratio, be made in the same way of negative controls.
2.3.4 the drafting of standard curve
It is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, makes 10.4,20.8,41.6,83.2,166.4 μ gmL with methanol
-1the solution of series concentration, precision measures each 10 μ L of above-mentioned 5 kinds of strength solution respectively, injects high performance liquid chromatograph and measures.Carry out linear regression with peak area ratio and concentration, obtaining regression equation is: A=21.2762C-0.1392, r=0.9998.Show that absinthol is at 10.2 ~ 166.6 μ gmL
-1in good linear relationship in concentration range.
2.3.5 stability test
Accurate absorption need testing solution 10 μ L, respectively at 0,1,2,4,8h sample introduction, and calculates absinthol content.In result 8h, RSD is 0.45%(n=5).Show that sample solution is stable in 8h.
2.3.6 replica test
By 5 parts, need testing solution preparation method parallel processing sample, measure absinthol content in accordance with the law and calculate.It is 0.12mgg that result records absinthol average content
-1, RSD is 1.3%.
2.3.7 Precision Experiment
Accurate absorption absinthol reference substance solution, repeats sample introduction 5 times, measures peak area in accordance with the law.Result RSD is 0.23%(n=5).Show that precision is better.
2.3.8 response rate experiment
Precision takes the sample of the same lot number of known absinthol content, and (lot number: 141016) 5 parts adds appropriate absinthol reference substance solution by high, medium and low concentration is accurate respectively, operates, measure in accordance with the law, calculate the response rate by under sample determination item.Result average recovery rate is 100.2%, RSD is 0.44%(n=5).
2.3.9 sample size measures
Measure reference substance solution and need testing solution respectively appropriate, filter with microporous filter membrane, each sample introduction 10 μ L, measures 3 batch samples by above-mentioned chromatographic condition, parallel assay 5 times.By external standard method with the content of calculated by peak area need testing solution absinthol.This product should be containing absinthol and indicates 95% ~ 105% of content, in every 1g sample containing absinthol, must not be less than 0.12mg.3 batch sample content are respectively 100.2%(RSD=1.1%), 101.7%(RSD=1.2%), 99.2%(RSD=1.2%).
detailed description of the invention:
embodiment 1:
Prescription: miliaria alba grass 250g, ferrum Brassaiopsis glomerulata (Bl.) Regel 250g, gristle wind 250g ， Radix Betae 250g.
Method for making: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin; put in spray-drying pelleting machine, make granule, add miliaria alba grass volatile oil; mixing, makes 1000g, is sub-packed in plastic bag; every bag of 5g, to obtain final product.
Detection method:
(1) chromatographic condition: adopt C18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg.
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
(5) testing result: count 0.14mg containing absinthol in every gram of Chinese medicine of the present invention
embodiment 2:
Prescription: miliaria alba grass 200g, ferrum Brassaiopsis glomerulata (Bl.) Regel 300g, gristle wind 200g ， Radix Betae 300g.
Method for making: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin; put in spray-drying pelleting machine, make granule, add miliaria alba grass volatile oil; mixing, makes 1000g, is sub-packed in plastic bag; every bag of 5g, to obtain final product.
Detection method:
(1) chromatographic condition: adopt C18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg.
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
(5) testing result: count 0.13mg containing absinthol in every gram of Chinese medicine of the present invention
embodiment 3:
Prescription: miliaria alba grass 300g, ferrum Brassaiopsis glomerulata (Bl.) Regel 300g, gristle wind 200g ， Radix Betae 200g.
Method for making: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin; put in spray-drying pelleting machine, make granule, add miliaria alba grass volatile oil; mixing, makes 1000g, is sub-packed in plastic bag; every bag of 5g, to obtain final product.
Detection method:
(1) chromatographic condition: adopt C18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg.
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
(5) testing result: count 0.15mg containing absinthol in every gram of Chinese medicine of the present invention
embodiment 4:
Prescription: miliaria alba grass 300g, ferrum Brassaiopsis glomerulata (Bl.) Regel 200g, gristle wind 300g ， Radix Betae 200g, Semen Vitis viniferae 10g, pueraria root powder 5g.
Method for making: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter, it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin; put in spray-drying pelleting machine, make granule, add miliaria alba grass volatile oil; mixing, makes 1000g, is sub-packed in plastic bag; every bag of 5g, to obtain final product.
Detection method:
(1) chromatographic condition: adopt C18(4.0mm × 125mm, 5 μm) chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg.
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
(5) testing result: count 0.17mg containing absinthol in every gram of Chinese medicine of the present invention.
Miliaria alba grass in Chinese medicine composition of the present invention is Labiatae Herba Moslae Scabrae platymiscium flower thinning Ji thuja
the herb of Mosla dianthera (Buch.-Ham.) Maxim. [M. Remotiflora Sun]; Ferrum Brassaiopsis glomerulata (Bl.) Regel is pulse family instrument flower plant instrument flower
lysidice rhodostegia Hanceroot; Gristle wind is Olacaceae Ramulus et Folium Schoepfiae chinesis platymiscium Ramulus et Folium Schoepfiae chinesis
schoepfia jasminodora Sieb. et Zucc. [Schoepfiopsis jasminodora (Sieb. et Zucc.) Miers]herb; Radix Betae is Chenopodiaceae Radix Betae platymiscium Chard dish
beta vulgaris L. var. cruenta Alef.and preserved carrot
b. vulgaris L. var. saccharifera Alef.root.
Claims (10)
1. treat a Chinese medicine composition for myocardial ischemia, it is characterized in that, this Chinese medicine composition is made up of the raw material of following weight portion: miliaria alba grass 1 ~ 5, ferrum Brassaiopsis glomerulata (Bl.) Regel 1 ~ 5, gristle wind 1 ~ 5, Radix Betae 1 ~ 5.
2. as claim 1 Chinese medicine composition, it is characterized in that, this Chinese medicine composition is made up of the raw material of following weight portion: miliaria alba grass 2, ferrum Brassaiopsis glomerulata (Bl.) Regel 3, gristle wind 2, Radix Betae 3.
3. as claim 1 Chinese medicine composition, it is characterized in that, this Chinese medicine composition is made up of the raw material of following weight portion: miliaria alba grass 3, ferrum Brassaiopsis glomerulata (Bl.) Regel 2, gristle wind 3, Radix Betae 2.
4. as claim 1 Chinese medicine composition, it is characterized in that, this Chinese medicine composition is made up of the raw material of following weight portion: miliaria alba grass 1, ferrum Brassaiopsis glomerulata (Bl.) Regel 1, gristle wind 1, Radix Betae 1.
5. as the Chinese medicine composition in Claims 1 to 4 as described in any one, it is characterized in that, this Chinese medicine composition adopts pharmaceutical methods conventional in pharmacy to be prepared into tablet, pill, hard capsule, granule, oral liquid.
6. Chinese medicine composition as claimed in claim 5, is characterized in that, this Chinese medicine composition is adopted and prepared with the following method: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 2 ~ 4 times, each 0.5 ~ 2h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 12 ~ 36h; filter; it is the clear paste of 1.00 ~ 1.10 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin, puts in spray-drying pelleting machine; make granule; add miliaria alba grass volatile oil, mixing, to obtain final product.
7. Chinese medicine composition as claimed in claim 6, is characterized in that, this Chinese medicine composition is adopted and prepared with the following method: heat-obtaining miliaria grass is placed in volatile oil extractor and distills, and extracts volatile oil, and gained miliaria alba grass volatile oil is for subsequent use; The another device of aqueous solution after the distillation of miliaria alba grass is collected, for subsequent use; Miliaria alba medical herbs slag after extraction volatile oil and ferrum Brassaiopsis glomerulata (Bl.) Regel, gristle Feng, Radix Betae decoct with water 3 times, each 1h, collecting decoction; filter, the aqueous solution after filtrate and miliaria alba grass distill merges, and staticly settles 24h; filter; it is the clear paste of 1.05 that filtrate is concentrated into 60 DEG C of relative densities, and qinghuo reagent adds cane sugar powder and dextrin, puts in spray-drying pelleting machine; make granule; add miliaria alba grass volatile oil, mixing, to obtain final product.
8. the Chinese medicine composition as described in Claims 1 to 4 any one, is characterized in that, the detection method of this Chinese medicine composition is:
(1) chromatographic condition: adopt C18 chromatographic column; Mobile phase: methanol-acetonitrile, ratio is 40:60; Determined wavelength: 238nm; Column temperature: 15 ~ 25 DEG C; Sensitivity: 0.1AUFS; Flow velocity: 1.0mLmin
-1; Sample size: 10 μ L;
(2) preparation of reference substance solution: it is appropriate that precision takes 80 DEG C of absinthol reference substances being dried to constant weight, adds methanol and makes the solution of every 1mL containing 0.2mg;
(3) preparation of need testing solution and negative controls: precision takes Chinese medicine granules 10g of the present invention, add methanol 40mL, reflux 4h, extracting solution reflux solvent is also concentrated into dry, residue add water 10mL dissolve, extract 5 times with water saturated n-butyl alcohol jolting, each 20mL, merge n-butanol extracting liquid, 3 times are washed with ammonia solution, each 15mL, n-butanol extracting liquid recycling design is to dry, and residue adds dissolve with methanol and is transferred in 10mL volumetric flask, add methanol to scale, shake up, filter, get filtrate and get final product;
(4) sample size measures: it is appropriate to measure reference substance solution and need testing solution respectively, filters, each sample introduction 10 μ L with microporous filter membrane.
9. the Chinese medicine composition as described in Claims 1 to 4 any one, is characterized in that, the application of this Chinese medicine composition in the Chinese medicine of preparation treatment myocardial ischemia.
10. the Chinese medicine composition as described in Claims 1 to 4 any one, is characterized in that, the application of this Chinese medicine composition in preparation treatment myocarditis Chinese medicine.
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Application publication date: 20150506 |