CN102973676B - Medicament for preventing and treating cardiovascular diseases and preparation method thereof - Google Patents
Medicament for preventing and treating cardiovascular diseases and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a medicament for preventing and treating cardiovascular diseases and a preparation method thereof. The medicament for preventing and treating cardiovascular diseases comprises total saponins of American ginseng stem and leaf and total saponins of red peony root. The preparation method comprises the following steps: preparation of total saponins of American ginseng stem and leaf: decocting American ginseng stem and leaf, filtering the decoctions, combining the filtrates, absorbing through macroporous resin, eluting with water, eluting with 50-70 volume percent ethanol, collecting eluate, and concentrating into extract with relative density between 1.06 and 1.08 at 80 DEG C; and preparation of the total saponins of red peony root: performing reflux extraction to red peony root with ethanol, combining filtrates, reclaiming ethanol under reduced pressure, concentrating, diluting with water, refrigerating for 20 to 28 hours at a temperature between 0 and 4 DEG C, filtering, adsorbing with macroporous resin, washing with water until the reducing sugar is negative, and eluting with ethanol. The medicament is simple in manufacture process and low in cost, is completely prepared from natural plants, does not contain hormone or pigment and other chemical synthetic products, and does not have toxic or side effect to a human body after being used continuously.
Description
Technical field
The present invention relates to Chinese medicine, particularly medicine of a kind of prevention and cure of cardiovascular disease and preparation method thereof.
Background technology
Cardiovascular disease, is called again blood circulation diseases, is the disease that a class relates to blood circulation, as: hypertension, coronary heart diseases and angina pectoris and acute myocardial infarction etc., serious caused exercise related sudden death.Relevant with the life style that arteriosclerosis, hyperlipemia, obesity, smoking etc. are bad.
Angina pectoris in cardiovascular disease belongs to the thoracic obstruction, the angina pectoris of the traditional Chinese medical science, the category of precordial pain with cold limbs.Primary disease is often with hypertension, hyperlipemia, atherosclerosis, diabetes etc.Chinese medicine thinks that coronary heart disease is the Primary Asthenia-Secondary Sthenia Syndrome of simulataneous insufficiency and excessive.Clinical manifestation is with the individual different very big differences that have, and when opinion is controlled, depending on change of illness state, its pathogenesis is stagnation of QI-blood, impatency breast sun, stagnation of QI and blood may bring about pain.
Symptomatic treatment in acute condition, radical treatment in chronic case, or treating both the principal and the secondary aspects of a disease at the same time, unfold the sun in ambition, and blood vessels operation is unimpeded.Effect a permanent cure and adopt the method for warming YANG and invigorating QI, nourishing YIN and benefiting blood; Take stopgap measures with cold relieving, eliminating phlegm, the method such as invigorate blood circulation.In a word, debate deficiency and excess, bright specimen is carried out tonify deficiency or removing the relative excess, or giving consideration to both the incidental and fundamental, carry out medicine typing treatment, just can obtain good effect.
Summary of the invention
For the deficiency existing in existing drug technique, the object of the present invention is to provide a kind of completely by natural plants make, compatibility is simple and the medicine of true medicable prevention and cure of cardiovascular disease, and provides the process for preparing medicine of the simple prevention and cure of cardiovascular disease of a kind of processing technology.
Technical scheme of the present invention is achieved in that a kind of medicine of prevention and cure of cardiovascular disease, contains stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides.
The medicine of above-mentioned prevention and cure of cardiovascular disease, the mass ratio of stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides is: stem and leaf of Radix Panacis Quinquefolii total saponins: Radix Paeoniae Rubra total glycosides=1:3～2:1.
The medicine of above-mentioned prevention and cure of cardiovascular disease, the mass ratio of stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides is: stem and leaf of Radix Panacis Quinquefolii total saponins: Radix Paeoniae Rubra total glycosides=1:2.
The preparation method of the medicine of above-mentioned prevention and cure of cardiovascular disease, produces respectively stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides;
Producing of stem and leaf of Radix Panacis Quinquefolii total saponins comprises the steps: to get stem and leaf of Radix Panacis Quinquefolii, decoct with water, decoct with water for the first time 1.5～2.5 hours, decoct with water for the second time 1～2 hour, decocting liquid filters, filtrate merges, by D101 macroporous adsorbent resin, water elution is colourless to eluent, then is 50～70% ethanol elutions by volume fraction, collects the eluent while being 50～70% ethanol elution by volume fraction, the extractum that while then eluent being concentrated into 80 ℃, relative density is 1.06～1.08, vacuum drying, pulverizes, and obtains stem and leaf of Radix Panacis Quinquefolii total saponins.
Producing of Radix Paeoniae Rubra total glycosides comprises the steps: to get Radix Paeoniae Rubra powder, with alcohol reflux, more than twice or twice, each 1～2 hour, filters merging filtrate; Decompression recycling ethanol, concentrated, thin up; Then be placed in 0～4 ℃ of cold preservation 20～28 hours, filter; Filtrate is by D101 macroporous resin adsorption, and first water rinses to reducing sugar in flushing liquor and is negative, then uses ethanol elution, and eluent reclaim under reduced pressure, obtains the thick cream of Radix Paeoniae Rubra extract; Thick cream adds ethyl acetate hot reflux, cooling after, filter, filtrate decompression is concentrated, obtains the extracting solution of Radix Paeoniae Rubra, vacuum drying, obtains Radix Paeoniae Rubra total glycosides.
Above-mentioned preparation method, producing of stem and leaf of Radix Panacis Quinquefolii total saponins comprises the steps: to get stem and leaf of Radix Panacis Quinquefolii decoction, decoct is that 1:8～12 decoct with water according to the mass ratio of stem and leaf of Radix Panacis Quinquefolii and water at every turn, decoct with water for the first time 2 hours, decoct with water for the second time 1.5 hours, decocting liquid filters, filtrate merges, by D101 macroporous adsorbent resin, first water elution is colourless to eluent, by volume fraction, be 60% ethanol elution again, collect the eluent while being 60% ethanol elution by volume fraction, the extractum that while being concentrated into 80 ℃, relative density is 1.06～1.08, vacuum drying, pulverize, obtain stem and leaf of Radix Panacis Quinquefolii total saponins.
Producing of Radix Paeoniae Rubra total glycosides comprises the steps: to get Radix Paeoniae Rubra powder, and the mass ratio of Radix Paeoniae Rubra powder and 70% ethanol of take is 1:6～10 reflux, extract, three times, each 1.5 hours, filters merging filtrate; Decompression recycling ethanol, is concentrated into every liter containing 1000 grams of crude drugs, adds the water dilution of 3 liters 60 ℃ according to every liter of concentrated solution; Then be placed in 0～4 ℃ of cold preservation 24 hours, filter, filtrate is by D101 macroporous resin adsorption, water rinses to reducing sugar in flushing liquor and is negative, using volume fraction instead is 30% ethanol elution, elution flow rate: 3mL/min, and eluant consumption is 3 times of amount resin column volumes, eluent reclaim under reduced pressure, obtains the thick cream of Radix Paeoniae Rubra extract; Thick cream adds ethyl acetate hot reflux, cooling after, filter, filtrate decompression is concentrated, obtains the extracting solution of Radix Paeoniae Rubra, vacuum drying, obtains Radix Paeoniae Rubra total glycosides.
The invention has the beneficial effects as follows:
The medicine of prevention and cure of cardiovascular disease of the present invention contains stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides, the two is made according to a certain percentage, can be made into granule, capsule, soft capsule, electuary, drop pill and soft extract, its processing technology is simple, and cost is low, by natural plants, is made completely, not containing hormone, do not add other synthetics such as pigment, after using continuously, human body is had no side effect yet.This Chinese medicine contains ginsenoside, the Radix Paeoniae Rubra total glycosides composition being easily absorbed by the body; can be effective to be applicable to the treatment of angina pectoris, acute myocardial infarction, cardiac function is had to protective effect, can resisting fatigue; the effect of enhancing immunity, has the effect of benefiting QI for activating blood circulation, blood-activating analgetic.
Stem and leaf of Radix Panacis Quinquefolii total saponins is dry stem and leaf by the Radix Panacis Quinquefolii total saponins through being processed into, micro-smelly, bitter in the mouth.The stem and leaf total saponins that Main Ingredients and Appearance is comprised of ginsenoside Rg1, Re, Rc, Rb2, Rd etc.The effects such as it has arrhythmia, resist myocardial ischemia, anti-oxidation stress, strengthening cardiac contractility ability.Can be used for the patients with coronary heart disease such as deficiency of both QI and YIN, palpitation and short breath, also can be for the treatment of the diseases such as hypertension, acute myocardial infarction, cerebral thrombosis.
Radix Paeoniae Rubra total glycosides is that Radix Paeoniae Rubra Chinese crude drug is through the total effective parts-Radix Paeoniae Rubra total glycosides (TotalPaeony Glycoside, TPG) of extraction, purification refine, mainly containing paeoniflorin, phenolic acid compound.Radix Paeoniae Rubra has clearing away heat and cooling blood, blood circulation promoting and blood stasis dispelling.Main maculae caused by violent heat pathogen, hematemesis and epistaxis, discharging fresh blood stool, conjunctival congestion and swelling pain etc.Modern pharmacology experimental results show that there is antiplatelet aggregation, antithrombotic, also can blood fat reducing and antiatherogenic effect, cardiovascular system is had to coronary artery dilator, increase the function of blood flow.Can be used for angina pectoris, the treatments such as the rehabilitation of acute myocardial infarction, hyperlipemia.
Stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides adopt macroporous resin adsorption isolation technics to extract and form, and compatibility forms in proportion.Have active constituent content high, yield is high, the curative effect feature such as clearly.
Function cures mainly: boosting qi and nourishing yin, blood circulation promoting and blood stasis dispelling.Be applicable to the Treating Patients of Angina Pectoris of type of deficiency of both QI and YIN, also can be used for acute myocardial infarction, hypertension, ARR auxiliary treatment, palpitation and short breath, atherosclerosis and cardiac insufficiency etc. are had to improvement effect.Disease sees that chest pain is uncomfortable in chest, shortness of breath and palpitation, spiritlessness and weakness, spontaneous perspiration, and color of the lip is purple dark, and purplish tongue is dark or have an ecchymosis, deficient and weak pulse or puckery, knot generation etc.
During < < Plain Questions visceral-qi method, opinion > > has: the discussion of " heart patient; chest pain; discomfort and fullness in the hypochondrium; pain over the hypochondrium; bear pain between back of the body omoplate; pain in the medial sides of both arms ", at < < Ling Shu Miraculous Pivot or Divine Axis, faint disease > > when coronary heart disease is sent out to angina pectoris attacks, description during to acute myocardial infarction: " angina pectoris; bluish colour of the limbs extending up to the elbows and knees; very pained, onset of disease in the morning and dying in the evening, onset of disease in the evening and dying in the next morning." common motive virtual loss or deficiency of both QI and YIN pattern of syndrome clinically, showing as moving tachypnea, spontaneous perspiration cardiopalmus, spiritlessness and weakness, pale complexion, sound are humble etc., and lip color is purple dark, and purplish tongue is dark or have an ecchymosis, deficient and weak pulse or puckery, knot generation etc.Acute stage,, expectorant, the stasis of blood, heat, the evil heap soil or fertilizer over and around the roots of poison were contained as mentioned above, and chronic phase is main mainly with retention of blood stasis.Accordingly, treatment should be take supplementing QI and nourishing YIN, warming YANG and invigorating QI, promoting blood circulation to remove blood stasis as main.
Stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides compatibility have the effect of warming YANG and invigorating QI, benefiting QI for activating blood circulation, yin nourishing blood stasis dispelling.Blood stasis due to qi deficiency or qi depression to blood stasis, phlegm stagnation in collateral is the main Chinese pathology basis of angina pectoris, myocardial infarction.Ancient Chinese medicine doctor application warming YANG and invigorating QI, activating blood and removing stasis Method are the Therapeutic Principle of the treatment thoracic obstruction, pained and angina pectoris.
Accompanying drawing explanation
Fig. 1 is rats in sham-operated group heart general pathology (HE).
Fig. 2 is model group rat heart general pathology (HE).
Fig. 3 is Western medicine group rat heart general pathology (HE).
Fig. 4 is Radix Panacis Quinquefolii Radix Paeoniae Rubra group rat heart general pathology (HE).
Fig. 5 is sham operated rats cardiac ultrasonic kinetocardiogram.
Fig. 6 is model group cardiac ultrasonic kinetocardiogram.
Fig. 7 is Western medicine group cardiac ultrasonic kinetocardiogram.
Fig. 8 is Radix Panacis Quinquefolii Radix Paeoniae Rubra group cardiac ultrasonic kinetocardiogram.
The specific embodiment
The extraction of stem and leaf of Radix Panacis Quinquefolii total saponins: get stem and leaf of Radix Panacis Quinquefolii, be cut into 1-2 centimetre of section, the decocting that medical material adds 10 times of amounts boils [decoct is that 1:10 decocts with water according to the mass ratio of stem and leaf of Radix Panacis Quinquefolii and water at every turn], decoct with water for the first time 2 hours, decoct with water for the second time 1.5 hours, decocting liquid filters, filtrate merges, by D101 macroporous adsorbent resin, first water elution is colourless to eluent, by volume fraction, be 60% ethanol elution again, collect the eluent while being 60% ethanol elution by volume fraction, and the extractum that relative density is 1.06～1.08 when eluent is concentrated into 80 ℃, vacuum drying, pulverize, obtain stem and leaf of Radix Panacis Quinquefolii total saponins, average yield is 3.5%.
The extraction of Radix Paeoniae Rubra total glycosides: get Radix Paeoniae Rubra powder, take mass ratio that Radix Paeoniae Rubra powder and volume fraction be 70% ethanol as 1:8 reflux, extract, three times, each 1.5 hours, filter merging filtrate.Decompression recycling ethanol, is concentrated into every liter containing 1000 grams of crude drugs [that is: every 1000 grams of Radix Paeoniae Rubra powder obtain concentrated solution 1L], adds the hot water dilution of 3 liters 60 ℃ according to every liter of concentrated solution.Then be placed in 0-4 ℃ of cold preservation 24 hours, filter, filtrate is by D101 macroporous resin adsorption, water rinses to reducing sugar in flushing liquor and is negative (5 times of amounts of water reducible resin bed volume), and using volume fraction instead is 30% ethanol elution, elution flow rate: 3mL/min, eluant consumption is 3 times of amount resin column volumes (volume ratio that is eluant and resin is 3:1), eluent reclaim under reduced pressure, obtains the thick cream of Radix Paeoniae Rubra extract, and average yield is 4.5%.。Thick cream adds ethyl acetate hot reflux, cooling after, filter, filtrate decompression is concentrated, obtains the extracting solution of Radix Paeoniae Rubra, vacuum drying, obtains Radix Paeoniae Rubra total glycosides, average yield is 3%.
Test a stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides compatibility to acute myocardial infarction of rat after cardiac muscle protective effect
1 materials and methods
1.1 drug study medications, stem and leaf of Radix Panacis Quinquefolii total saponins 50mg, Radix Paeoniae Rubra total glycosides 100mg.Western medicine group be valsartan capsule (trade name: DAIWEN), 80mg/ grain, lot number: X1092, the accurate word of traditional Chinese medicines: H20040217, is produced by Novartis Pharma AG.
1.2 140 of animal health Wistar rats, clean level, male and female half and half, body weight (200-220) g, is provided the quality certification number by Beijing Vital River Experimental Animals Technology Co., Ltd.: SCXK(capital) 2007-0001.Rat is raised in clean level animal housing after purchasing, indoor temperature (23～25) ℃, humidity (50%～70%), animal ad lib and take the photograph water, illumination and dark each 12h.Animal adaptability is tested after raising 7d.
1.3 reagent Type B brain natriuretic peptide (BNP) radioimmunological kits, Beijing China English biotechnology research provides, lot number: 20120115; Cardiac troponin T (cTNT) radioimmunological kit, Beijing China English biotechnology research provides, lot number: 20120128.
1.4 instrument ECG-6511 type electrocardiographs, Shanghai photoelectric instrument company limited; Olympus BH-2 polarizing microscope, Japanese Olympus company; 80-2 type centrifuge, Shanghai Surgical Operation Equipment Factory; R-911 type automatically radiates immune gamma counter, and industry head office of Chinese University of Science and Technology produces; HDI5000 type diasonograph and CL15-7 type high-frequency linear array (7-15MHZ) probe, Philip (China) Investment Co., Ltd. produces.
1.5 method
1.5.1 model preparation experiment rat, 4% chloral hydrate intraperitoneal injection of anesthesia (0.1mL/kg body weight) is fixed on its dorsal position on animal operation table after animal loses autonomic activities ability.Preserved skin, sterile surgical is wild, determines the strength of apex beat, and left border of sternum 3-4 intercostal is longitudinally cut off skin, and successively blunt separation subcutaneous tissue and thoracic wall muscle, to exposing rib and intercostal space.With curved forceps, scratch gently intercostal space, expose heart, cut off pericardium, with 3/8 circle 2 * 6 swaged needles, 1/3 place's projected position threading ligation blood vessel on ramus descendens anterior arteriae coronariae sinistrae, to block coronary blood flow.Stablize 2min, with ECG-6511 electrocardiograph, record II lead electrocardiogram, with electrocardiogram occur ST section back of a bow sample raise (with) or T wave height alarm, it is ligation success that ligation blood vessel blood supply district cardiac muscle is kermesinus.Send rapidly the heart after ligation back to thoracic cavity, extrude air in thoracic cavity, layer-by-layer suture is closed otch.Postoperative muscle injection penicillin sodium 200,000 U/d, for three days on end prevention infection.Only not ligation of threading of sham operated rats, the rat that after modeling, success in 24 hours is survived enters to organize experimentation.
1.5.2 experiment grouping and administration are divided into 6 groups by the Wistar rat that after modeling, success is survived random (table of random number), and 15 every group, male and female half and half.Grouping and administration are as follows: 1. sham operated rats: gavage normal saline 0.2mL/ (kgd); 2. model group: gavage normal saline 0.2mL/ (kgd); 3. Western medicine group: give valsartan rat dosage gavage: 7.2mg/kgd; 4. Radix Panacis Quinquefolii Radix Paeoniae Rubra group: (mass ratio is stem and leaf of Radix Panacis Quinquefolii total saponins: Radix Paeoniae Rubra total glycosides=1:2) to give stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides.
Trial medicine is all by the same equimultiple of distilled water diluting, by 0.1mL/kg body weight gavage.Sham operated rats and model group often daily give the distilled water gavage of equivalent, continuously gastric infusion 28d.After rat AMI modelling completes, in gastric infusion process, rat is total to dead 8.Wherein sham operated rats is dead 1, each 2 of model group and Western medicine groups, dead 3 of Radix Panacis Quinquefolii Radix Paeoniae Rubra group.Analyze the cause of death and be due to heart failure, pulmonary infection and the gavage misoperation after heart infarction.
1.5.3 each treated animal gavage of echocardiography 28d, during 28d, water 12h is can't help in fasting.29d, 4% chloral hydrate intraperitoneal anesthesia (0.1mL/kg body weight).With 8% sodium sulfide solution, to the depilation of rat front, the successful rat dorsal position of preserved skin is fixed on to the up Cardiac ultrasound of operation plate.Probe is placed in its left breast, after obtaining satisfied parasternal left-ventricular short-axis two dimensional image, at mitral level, M type sample line is obtained to M type ultrasoundcardiogram perpendicular to interventricular septum and left ventricular posterior wall.Ultrasoundcardiogram measurement index comprises: left chamber end systolic diameter (the 1eft ventricular end-systoledimension of same cardiac cycle, LVDs), LVED (Left Ventricular End Systolic Dimension) (1eft ventricular end-diastoledimension, LVDd), interventricular septum end-diastolicthickness (interventricular septumend-diastole thickness, IVSTd), interventricular septum end-systole thickness (interventricularseptum end-systole thickness, IVSTs), LVPW end-diastolicthickness (1eftventricular posterior wall end-diastole thickness, LVPWTd), LVPW end-systole thickness (1eft ventricular posterior end-systole thickness, LVPWTs), Left Ventricular Ejection Fraction (1eft ventricular ejection fraction, LVEF).The meansigma methods of 3 cardiac cycles of every group of initial data METHOD FOR CONTINUOUS DETERMINATION during measurement, ultrasound procedure personnel and analyst all do not know rat grouping situation (single blind).
1.5.4 serology detects after ultrasound detection finishes, abdominal aortic blood, and separation of serum-80 ℃ Refrigerator store is to be measured.The mensuration of serum cTNT, BNP adopts radioimmunology, and the mensuration of inflammatory factor TNF-α, IL-1 β adopts radioimmunology.
1.5.5 pathomorphology detects and gets after blood, and every group of rat chosen 5 at random, extracts heart, and normal saline flushing, puts into 10% neutral formalin solution and preserve after filter paper suck dry moisture, in order to pathological observation.Rat heart muscle structure is observed in conventional embedding, dewaxing, section, dyeing, haematoxylin-Yihong (hematoxylinand eosin, HE) dyeing, the morphosis of normal optical Microscopic observation rat heart muscle tissue.
1.5.6 statistical method
All data are processed by SPSSl7.0 statistical software, with mean ± standard deviation
represent, while meeting normal distribution and homogeneity of variance, with one factor analysis of variance (ONE-WAY ANOVA), further between group, multiple comparisons adopts LSD method.Do not meet homogeneity of variance and by Welch method, F value is proofreaied and correct, further Dunnett T3 method for multiple comparisons, the P < 0.05 of take has statistical significance as difference.
2 results
2.1 respectively organize rat heart structure (heart general pathology) and cardiac muscular tissue's pathological change
The visible ventricle wall of sham operated rats heart is normal, and the chambers of the heart is without remarkable expansion, and morphosis is normal.The remarkable attenuation of the visible locular wall of model group, the chambers of the heart expands, whole ventricular chamber expansion, distortion.The animal locular wall attenuation of valsartan, Radix Panacis Quinquefolii Radix Paeoniae Rubra group and chamber expand situation and all compared with model group, alleviate.
Sham operated rats myocardial cell marshalling, without degeneration, intercellular substance is without edema, hemorrhage, cardiac muscular tissue's muscle fiber band and nucleus dyeing are clear.There is extensive amalgamation pathological changes in model group cardiac muscle, myolin arrangement disorder, and part is corrugated and changes, part muscle fiber liquefacttus, myocardium interstitial can occur that inflammatory cell invades profit; Myocardial cell karyopycnosis, cracked or dissolving disappearance, part cardiac muscle is kitchen range shape and lamellar is downright bad.Infarcted region karyopycnosis or dissolving after animal model, cardiac muscle fiber is red dyes, gap broadening, band disappears, and fracture is remarkable, and has the degeneration of part cardiac muscle fiber sample, and edge, infarcted region is congested as seen, hemorrhage band and inequality neutrophil infiltration.The myocardial cell inflammatory infiltration of valsartan, Radix Panacis Quinquefolii Radix Paeoniae Rubra treated animal reduces, and henle's fissures edema alleviates significantly, and the degeneration of cardiac muscle fiber necrocytosis is alleviated, and Radix Panacis Quinquefolii Radix Paeoniae Rubra group is comparatively remarkable.See accompanying drawing 1-4.
2.2 respectively organize Cardiac Function in Rat detects
Echocardiography shows after heart infarction modeling 4 weeks, and rat left chamber expands, and the contractile function of whole heart reduces.With sham operated rats comparison, LVDd, the LVDs of model group and each administration group rat (the P < 0.05 that all significantly raises, P < 0.01), IVSTd, IVSTs, LVEF all significantly reduce (P < 0.01), prompting modeling success, conforms to research report in the past.With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group rat LVDd all significantly reduces (P < 0.05), and LVDs reduces significantly (P < 0.05).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group IVSTd, IVSTs be significantly rising (P < 0.05) all.With sham operated rats comparison, model and Radix Panacis Quinquefolii Radix Paeoniae Rubra group rat LVEF all significantly reduce (P < 0.01).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group rat LVEF is significantly rising (P < 0.05) all.In Table 1 and Fig. 5-8.
Table 1 is respectively organized Cardiac Function in Rat comparison
Note: with sham operated rats comparison,
*p < 0.05,
*p < 0.01; With model group comparison,
△p < 0.05,
△ △p < 0.01
2.3 respectively organize rat blood serum cTnT, BNP level
Each is organized rat blood serum myocardial injury markers cTnT content and relatively has no significant difference (P > 0.05), and 4 weeks rear serum cTnT levels of prompting modeling have recovered normal.With sham operated rats comparison, model group and Radix Panacis Quinquefolii Radix Paeoniae Rubra group rat blood serum BNP content significantly raise (P < 0.01).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group serum BNP level significantly reduces (P < 0.05), though all the other two groups of serum BNP levels have reduction, not statistically significant (P > 0.05).In Table 2.
Each administration group impact on rat blood serum cTnT, BNP of table 2
| Group | N | cTnT(ng/mL) | BNP(pg/mL) |
| Sham operated rats | 14 | 0.041±0.03 | 8.52±1.81 △△ |
| Model group | 13 | 0.046±0.03 | 16.29±1.99 ** |
| Western medicine group | 13 | 0.037±0.02 | 14.90±1.92 **△ |
| Radix Panacis Quinquefolii Radix Paeoniae Rubra group | 12 | 0.034±0.01 | 14.58±1.35 **△ |
Note: with sham operated rats comparison,
*p < 0.05,
*p < 0.01; With model group comparison,
△p < 0.05,
△ △p < 0.01
3 conclusions
After acute myocardial infarction (AMI), the prevention of heart failure is an important step can not be ignored.The generation of early stage remodeling ventricle (VR), main manifestations is the swollen exhibition of infarcted region cardiac muscle, locular wall attenuation, chamber expansion.Think that at present the main pathological basis that chronic heart failure occurs after heart infarction is remodeling ventricle, remodeling ventricle is considered to the sign of heart failure long-term prognosis and is the target spot of control.4 weeks rear rat heart general structures of modeling have been observed in this research, find that the attenuation of model group heart chamber wall, chamber expand, and the morphosis that meets VR changes.The Pathologic Characteristics of VR comprises myocardial cell phenotypic alternation, cardiomyocyte cell death, and fibrosis, blood vessel lacks, cardiac myocyte hypertrophy, mitochondrion lacks, calcium ion transport obstacle, myosin bradykinesia etc.This research finds that after 4 weeks, medication therapy groups myocardial cell inflammatory infiltration significantly reduces, and henle's fissures edema significantly alleviates, karyopyknosis, cracked or dissolution phenomena minimizing.
Ultrasoundcardiogram is commonly used to objective evaluation cardiac function clinically, and result of study shows after AMI 4 weeks, and at tangent planes such as the left chamber of model group parasternal major axis, apical four-chambers, all visible left chamber antetheca, front side wall all have outwards bulging in various degree, locular wall attenuation.Infarcted region is without shrinking or systole adverse movement, the remarkable bulge of apex, and left ventricular interior diameter significantly expands simultaneously, the generation proof modeling success of prompting VR.After taking the medicine of stem and leaf of Radix Panacis Quinquefolii total saponins Radix Paeoniae Rubra total glycosides compatibility, rat heart muscle shrinks and ED left ventricular interior diameter and significantly reductions of model group, Left Ventricular Ejection Fraction is also significantly improved simultaneously, and after the treatment of prompting drug application, the cardiac function of rat improves.
Serum cardiac troponin T (cardiac troponin T, cTNT) be the blood serum designated object for reflecting myocardium necrosis of generally acknowledging, result of study showed after 4 weeks, it is all lower that each organizes rat blood serum cTnT level, between group, compare there was no significant difference, the acute stage of prompting cardiac damage, is mistake, and after 4 weeks, serum cTnT has been down to normally.BNP (B-type natriuretic peptide, BNP) be a kind of sharp natriurtic peptide of mainly being secreted by ventricular muscle cell, result of study shows, heart infarction modeling after 4 weeks model group and each administration group serum BNP level all significantly raise and point out cardiac insufficiency after heart infarction and the generation of VR.After taking the medicine of stem and leaf of Radix Panacis Quinquefolii total saponins Radix Paeoniae Rubra total glycosides compatibility, BNP level all has decline compared with model group.Illustrate after Drug therapy, rat left chamber's pressure and locular wall tension force reduce, and ventricular dilatation alleviates, and cardiac function improves.
Test two stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides compatibility on acute myocardial infarction of rat after the impact of inflammatory factor and extracellular matrix
1 materials and methods
1.1 medicines and animal are with experiment one.
1.2 reagent: tumor necrosis factor-alpha (TNF-α) radioimmunological kit, lot number: 20120125; Interleukin-1 ' beta ' (IL-1 β) radioimmunological kit, lot number: 20120123; Laminin (LN) radioimmunological kit, lot number: 20120122; HA radioimmunological kit, lot number: 20120128; III procollagen type ammonia end peptide (P III NP) radioimmunological kit, lot number: 20120127; AngiotensionⅡ (Ang II) radioimmunological kit, lot number: 20120129; Aldosterone (ALD) radioimmunological kit, lot number: 20120125; Transforming factor-β 1(TGF-β 1) radioimmunological kit, lot number: 20120125.Above test kit provides by Beijing China English biotechnology research.
Collagen I antibody, lot number: 990184W; Collagen III antibody, lot number: 909881W; Metalloproteases-the 9(MMP-9 of periplast) antibody, lot number: 990394W; TIMP-1 (TIMP-1) antibody, lot number: 999999W; Osteopontin (OPN) antibody, lot number: 909984W; Tendon glycoprotein-C(TN-C) antibody, lot number: 900196W; Primary antibodie all derives from rabbit, and above antibody is provided by Beijing Bo Aosen Bioisystech Co., Ltd.The anti-goat IgG antibody-HRP of rabbit polymer, concentrated type diaminobenzidine test kit (Diaminobenzidine, DAB), by Beijing, Bioisystech Co., Ltd of Zhong Shan Golden Bridge provides.
1.3 instrument Olympus BH-2 polarizing microscopes, Japanese Olympus company; T18 high speed dispersor, German IKA company; R-911 type automatically radiates immune gamma counter, industry head office of Chinese University of Science and Technology; 7020 type full automatic biochemical apparatus, HIT; The micro-chromatic image processing system of DpxView Pro type, Denmark DeltaPix company.
1.4 method
The making of 1.4.1 animal grouping, administration and model is with experiment one.
1.4.2 every group of rat of cardiac bistiocyte's factors check chosen 6 hearts at random, get under anterior descending branch ligature 2mm to the left chamber of apex ischemic myocardial tissue 300mg, add 1mL low-temperature normal saline (0 ℃), after the homogenate of T18 high speed dispersor, centrifuging and taking supernatant, to be measured in-80 ℃ of preservations.
1.4.3 immunohistochemistry detects and utilizes abiotic plain two step method to carry out immunohistochemical detection, and the formaldehyde that myocardium specimen configures with 10% neutral buffered liquid is 18h fixedly.Dehydration, transparent, waxdip, paraffin embedding conventional treatment.Approximately 5 μ m are thick for serial section, drag on the microscope slide that sheet processed in poly-D-lysine anticreep.According to abiotic plain two step method test kit description operation, step is as follows: rat heart muscle section is conventional de-cured, pretreatment; 3%H2O2 deionized water is hatched 5-10min, blocking-up endogenous peroxydase, and phosphate buffer (PBS) rinses, 2min * 3 time; Drip respectively primary antibodie TN-C receptor antibody (dilution ratio, 1:300, OPN receptor antibody (1:300), Collagen I receptor antibody (1:300), collagen III receptor antibody (1:300), TIMP-1 receptor antibody (1:400), MMP-9 receptor antibody (1:300) down together),, hatch 1h for 37 ℃, PBS rinses, 2min * 3 time; Drip the anti-goat IgG antibody-HRP of rabbit polymer, incubated at room 30min, PBS solution rinses, 2min * 3 time; Select DBA colour developing, micro-Microscopic observation, controls the response time under mirror, sucks nitrite ion when organizing painted and back end paler colour at once; Distilled water fully rinses, and haematoxylin is slightly redyed, ethanol dehydration, transparent, neutral gum mounting.
Choose with batch micro-Microscopic observation of stained.3 continuous visuals field (* 600) are chosen in every section at random, every amounts to 15 visuals field, with the endochylema at myocardial damage position, dye the positive cell sign of sepia, adopt the professional pathological image analysis software of Imagepro-Plus 6.0 to measure integral optical density value (the integrated optical density in cardiac muscular tissue's section positive expression district, IOD), average as IOD value.
1.4.4 statistical method is with experiment one.
2 results
2.1 respectively organize variation and the sham operated rats comparison of rat blood serum inflammatory factor level, and each organizes rat blood serum inflammatory factor TNF-alpha content significantly raise (P < 0.05, P < 0.01); With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group TNF-alpha content significantly reduces (P < 0.05).With sham operated rats comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group serum inflammatory factors of senile IL-1 β content significantly raise (P < 0.05, P < 0.01).In Table 3.
The impact of table 3. Radix Panacis Quinquefolii Radix Paeoniae Rubra group on rat blood serum inflammatory factor level
| Group | N | TNF-α(pg/mL) | IL-1β(ng/mL) |
| Sham operated rats | 14 | 1.18±0.18 △△ | 0.27±0.05 △△ |
| Model group | 13 | 1.81±0.49 ** | 0.49±0.08 ** |
| Western medicine group | 13 | 1.52±0.41 **△ | 0.41±0.10 **△ |
| Radix Panacis Quinquefolii Radix Paeoniae Rubra group | 12 | 1.49±0.28 *△ | 0.33±0.11 △△ |
Note: with sham operated rats comparison,
*p < 0.05,
*p < 0.01; With model group comparison,
△p < 0.05,
△ △p < 0.01
2.2 respectively organize variation and the sham operated rats comparison of rat blood serum fibrosis factor level, and each organizes rat blood serum LN level significantly raise (P < 0.05).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group LN level significantly reduces (P < 0.05).With sham operated rats comparison, each organizes HA level significantly raise (P < 0.01).With model group comparison, stem and leaf of Radix Panacis Quinquefolii total saponins Radix Paeoniae Rubra total glycosides group HA level significantly reduces (P < 0.05), though Western medicine group HA level has reduction, and not statistically significant (P > 0.05).With sham operated rats comparison, each organizes P III NP level significantly raise (P < 0.01).With model group comparison, Western medicine group, Radix Panacis Quinquefolii Radix Paeoniae Rubra group P III NP level significantly reduce (P < 0.05).The results are shown in Table 4.
The impact of table 4. Radix Panacis Quinquefolii Radix Paeoniae Rubra group on rat blood serum fibrosis factor level
Note: with sham operated rats comparison,
*p < 0.05,
*p < 0.01; With model group comparison,
△p < 0.05,
△ △p < 0.01
2.3 respectively organize variation and the sham operated rats comparison that rat heart muscle is organized Ang II, ALD, TGF-β 1 content, and each is organized rat heart muscle and organizes Ang II content significantly raise (P < 0.05, P < 0.01).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group Ang II content significantly reduces (P < 0.01).With sham operated rats comparison, each is organized rat heart muscle and organizes ALD content (the P < 0.05 that significantly raises, P < 0.01), with model group comparison, the ALD content of Radix Panacis Quinquefolii Radix Paeoniae Rubra group all significantly reduces (P < 0.01).With sham operated rats comparison, each organizes rat heart muscle tissue T GF-β 1 content significantly raise (P < 0.05, P < 0.01).With model group comparison, Western medicine group and Radix Panacis Quinquefolii Radix Paeoniae Rubra group TGF-β 1 content significantly reduce (P < 0.01).The results are shown in Table 5.
Table 5. Radix Panacis Quinquefolii Radix Paeoniae Rubra group is organized the impact of Ang II, ALD, TGF-β 1 content on rat heart muscle
| Group | n | AngⅡ(pg/mL) | ALD(pg/mL) | TGF-β1(ng/L) |
| Sham operated rats | 6 | 34.66±5.61 △△ | 6.42±1.25 △△ | 51.15±9.20 △△ |
| Model group | 6 | 56.54±7.81 ** | 10.78±1.54 ** | 86.44±11.40 ** |
| Western medicine group | 6 | 50.20±3.50 **△ | 8.30±1.06 *△△ | 71.01±9.55 **△ |
| Radix Panacis Quinquefolii Radix Paeoniae Rubra group | 6 | 42.89±7.18 *△△ | 8.12±1.50 *△△ | 64.81±9.77 *△△ |
Note: with sham operated rats comparison, * P < 0.05, * * P < 0.01; With model group comparison, △ P < 0.05, △ △ P < 0.01
2.3 respectively organize rat heart muscle organizes the variation I of I, III collagen content, III Collagen Type VI to be positioned myocardial cell interstitial, expresses the positive and is brown color.In sham operated rats cardiac muscular tissue, the expression of type Ⅰ and Ⅲ collagen is very faint.With model group comparison, Western medicine group and Radix Panacis Quinquefolii Radix Paeoniae Rubra group Collagen I are expressed and are significantly reduced (P < 0.05).With model group comparison, Western medicine group, Radix Panacis Quinquefolii Radix Paeoniae Rubra group collagen III are expressed and are significantly reduced (P < 0.05, P < 0.01).The results are shown in Table 6.
The impact that table 6. Radix Panacis Quinquefolii Radix Paeoniae Rubra group is expressed rat heart muscle tissue collagen I, III
Note: with sham operated rats comparison, * P < 0.05, * * P < 0.01; With model group comparison, △ P < 0.05, △ △ P < 0.01
2.5 respectively organize rat heart muscle organizes variation OPN and the TN-C of OPN, TN-C expression to be all expressed in myocardial cell interstitial, and positive expression is that granule palm fibre is dyed.The accidental brown particle of sham operated rats myocardial cell interstitial.With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group OPN expresses and significantly reduces (P < 0.01).With model group comparison, Radix Panacis Quinquefolii Radix Paeoniae Rubra group TN-C expresses and significantly reduces (P < 0.05, P < 0.01), though Western medicine group has the not statistically significant (P > 0.05) of reduction.The results are shown in Table 7.
The impact that table 7. Radix Panacis Quinquefolii Radix Paeoniae Rubra group organizes OPN and TN-C to express on rat heart muscle
| Group | n | OPN | TN-C |
| Sham operated rats | 6 | 802.80±105.41 △△ | 875.80±279.98 △△ |
| Model group | 6 | 4138.60±994.25 ** | 2052.80±529.74 ** |
| Western medicine group | 6 | 3671.60±411.28 ** | 1709.20±438.89 ** |
| Radix Panacis Quinquefolii Radix Paeoniae Rubra group | 6 | 2924.80±658.36 **△△ | 1505.80±278.33 *△ |
Note: with sham operated rats comparison,
*p < 0.05,
*p < 0.01; With model group comparison,
△p < 0.05,
△ △p < 0.01
3 discuss
3.1 Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibility Chinese medicines are repaired in the process of reconstruct the immune activation state with multiple inflammatory cytokine to affect AMI and the myocardial damage of inflammatory factor, cause a series of relevant inflammatory reactions, wherein TNF-α, IL-1 β are one of cytokines of Main Function, they can with the receptors bind on various kinds of cell surface, and then start recruitment and the activation that cascade reaction causes neutrophilic granulocyte, mononuclear cell, macrophage, cause structural deterioration and the dysfunction of myocardial cell.This result of study shows, the heart infarction modeling visible significant hyperemia in rear Microscopic observation rat heart muscle pathology discovery edge, infarcted region in 4 weeks, hemorrhage band, and cardiac muscular tissue's inflammatory cell infiltration is remarkable.Detection by quantitative serum inflammatory factors of senile TNF-α, IL-1 β level significantly raise, after the treatment of Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibility, serum inflammatory factors of senile level significantly reduces, results suggest medicine may be by reducing the inflammatory reaction after heart infarction, the release that reduces inflammatory factor further reduces the expression of MMPs, alleviates VR.
The extracellular matrix that affects cardiac muscle (ECM) main component that 3.2 Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibilities form collagen is collagen fiber, and heart collagenous network is to maintaining cardiac structure and functional completeness plays a very important role.Especially the hypertrophy of NTx, makes I type and III Collagen Type VI ratio become large.After myocardial infarction, the change of Collagen I/III ratio has reflected the reconstruction of collagen grid structure.After the modeling of this result of study demonstration heart infarction, model group and administration group rat heart muscle type Ⅰ and Ⅲ collagen all have remarkable expression, and the expression of Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibility group significantly reduces compared with model group, the collagen that prompting Chinese medicine can reduce after heart infarction is synthetic, thereby alleviate myocardial collagen, piles up, and alleviates ECM reconstruct.
Although 3.3 Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibilities are considered to the outstanding feature thing in fibrotic processes to the impact collagen of myocardial fibrosis, remaining ECM albumen has also been brought into play important function in the character of determining ECM.This research discovery, in 4 weeks rear myocardium tissues of heart infarction, fibrosis factor level significantly raises, consistent with bibliographical information.The rising of fibrosis factor level has reflected the formation of myocardial fibrosis, and the generation of heart failure.Give after Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibility Chinese medicine, in serum, the level of the fibrosis factor significantly reduces, and shows that medicine can be reduced to fibrocellular hypertrophy, reduces its activity, alleviates the heart failure that cardiac damage and VR cause.
3.4 Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibilities also provide remaining important regulatory factor in cicatrization on the fact that heart ECM is comprised of different albumen that affects of OPN, TN-C in the adaptive while of having explained 26S Proteasome Structure and Function.OPN is a kind of important functional protein in the ECM in recent years finding, because its mediation bone and its cells is connected with bone matrix, participates in bone matrix mineralising and gains the name with heavy absorption process.On Rat of Myocardial Infarction model, observe left anterior descending coronary artery ligation after 4 weeks, the OPN of cardiac muscular tissue expresses significantly to be increased.Rat occurs that significant collagen fiber deposition, left ventricle dilatation, non-infarcted region cardiac myocyte hypertrophy and cardiac function decline simultaneously.The demonstration of this result of study, coronary ligation is respectively organized rat heart muscle OPN and is expressed significantly.Compare with model group, Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibility group OPN expresses significantly to reduce and points out OPN can promote the VR after heart infarction as ECM important composition albumen, and medicine can be by reducing generation and the expression inhibiting ECM reconstruct of OPN.
Tendon glycoprotein is to have highly bioactive ECM glycoprotein, and first the important member in TN-C Shi Jian glycoprotein family finds its expression in fetal development in Ye Shigai family.Originally studies show that, after modeling, the expression of rat heart muscle TN-C significantly increases, and after drug application treatment, the expression of TN-C reduces, and wherein the expression of Radix Panacis Quinquefolii and Radix Paeoniae Rubra compatibility group reduces more remarkable.And the reconstruct that medicine can alleviate ECM by reducing the generation of TN-C.
3.5 Radix Panacis Quinquefolii Radix Paeoniae Rubra compatibilities play a significant role in the physiology of cardiovascular system and pathology are replied on the RAAS that affects of neuroendocrine factor, wherein Ang II not only mediates vasoconstriction and regulates blood pressure as its main component, and increasing evidence its in congestive heart failure and inflammation damnification, play the part of important role.Ang II has the factor of TGF-β 1 to participate in the mechanism of cardiac fibroblast effect, and it is the key that Ang II causes myocardial collagen remodeling with activating that TGF-β 1 generates increase.After this research application Radix Panacis Quinquefolii and the treatment of Radix Paeoniae Rubra compatibe drug, Ang II, ALD, TGF-β 1 content all significantly reduce, and prompting medicine may be also by blocking-up RAAS, to reduce generation and TGF-β 1 secretion of Ang II.
In a word, stem and leaf of Radix Panacis Quinquefolii total saponins Radix Paeoniae Rubra total glycosides compatibility can improve the early stage remodeling ventricle after rat AMI, improve cardiac ejection fraction, improve cardiac function, its mechanism may relate to inflammation-inhibiting reaction, reduces the activation of neuroendocrine factor, regulate extracellular matrix metabolism relevant, there is control acute myocardial infarction, reduce myocardial infarction area, improve the effect of cardiac function.
Above-described embodiment is only for the invention example is clearly described, and the not restriction to the invention specific embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here exhaustive without also giving all embodiments.And the apparent variation of being amplified out thus or change are still among the protection domain in the invention claim.
Claims (2)
1. a medicine for prevention and cure of cardiovascular disease, is characterized in that, by stem and leaf of Radix Panacis Quinquefolii total saponins and Radix Paeoniae Rubra total glycosides, according to the ratio of 2:1, formed, wherein,
Described stem and leaf of Radix Panacis Quinquefolii total saponins is to be produced and obtained by following method: get stem and leaf of Radix Panacis Quinquefolii and decoct, decoct is that 1:8～12 decoct with water according to the mass ratio of stem and leaf of Radix Panacis Quinquefolii and water at every turn, decoct with water for the first time 2 hours, decoct with water for the second time 1.5 hours, decocting liquid filters, filtrate merges, by D101 macroporous adsorbent resin, first water elution is colourless to eluent, by volume fraction, be 60% ethanol elution again, collect the eluent while being 60% ethanol elution by volume fraction, the extractum that while being concentrated into 80 ℃, relative density is 1.06～1.08, vacuum drying, pulverize, obtain stem and leaf of Radix Panacis Quinquefolii total saponins,
Described Radix Paeoniae Rubra total glycosides is to be produced and obtained by following method:
Get Radix Paeoniae Rubra powder, take mass ratio that Radix Paeoniae Rubra powder and volume fraction be 70% ethanol as 1:6～10 reflux, extract, three times, each 1.5 hours, filter merging filtrate; Decompression recycling ethanol, is concentrated into every liter containing 1000 grams of crude drugs, adds the water dilution of 3 liters 60 ℃ according to every liter of concentrated solution; Then be placed in 0～4 ℃ of cold preservation 24 hours, filter, filtrate is by D101 macroporous resin adsorption, water rinses to reducing sugar in flushing liquor and is negative, using volume fraction instead is 30% ethanol elution, elution flow rate: 3mL/min, and eluant consumption is 3 times of amount resin column volumes, eluent reclaim under reduced pressure, obtains the thick cream of Radix Paeoniae Rubra extract; Thick cream adds ethyl acetate hot reflux, cooling after, filter, filtrate decompression is concentrated, obtains the extracting solution of Radix Paeoniae Rubra, vacuum drying, obtains Radix Paeoniae Rubra total glycosides.
2. the preparation method of the medicine of prevention and cure of cardiovascular disease described in a claim 1, it is characterized in that, producing of stem and leaf of Radix Panacis Quinquefolii total saponins comprises the steps: to get stem and leaf of Radix Panacis Quinquefolii decoction, decoct is that 1:8～12 decoct with water according to the mass ratio of stem and leaf of Radix Panacis Quinquefolii and water at every turn, decoct with water for the first time 2 hours, decoct with water for the second time 1.5 hours, decocting liquid filters, filtrate merges, by D101 macroporous adsorbent resin, first water elution is colourless to eluent, by volume fraction, be 60% ethanol elution again, collect the eluent while being 60% ethanol elution by volume fraction, the extractum that while being concentrated into 80 ℃, relative density is 1.06～1.08, vacuum drying, pulverize, obtain stem and leaf of Radix Panacis Quinquefolii total saponins,
Producing of Radix Paeoniae Rubra total glycosides comprises the steps: to get Radix Paeoniae Rubra powder, take mass ratio that Radix Paeoniae Rubra powder and volume fraction be 70% ethanol as 1:6～10 reflux, extract, three times, each 1.5 hours, filters merging filtrate; Decompression recycling ethanol, is concentrated into every liter containing 1000 grams of crude drugs, adds the water dilution of 3 liters 60 ℃ according to every liter of concentrated solution; Then be placed in 0～4 ℃ of cold preservation 24 hours, filter, filtrate is by D101 macroporous resin adsorption, water rinses to reducing sugar in flushing liquor and is negative, using volume fraction instead is 30% ethanol elution, elution flow rate: 3mL/min, and eluant consumption is 3 times of amount resin column volumes, eluent reclaim under reduced pressure, obtains the thick cream of Radix Paeoniae Rubra extract; Thick cream adds ethyl acetate hot reflux, cooling after, filter, filtrate decompression is concentrated, obtains the extracting solution of Radix Paeoniae Rubra, vacuum drying, obtains Radix Paeoniae Rubra total glycosides.
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