CN101618176A - Chinese medicament for treating dyslipidemia and atherosclerosis - Google Patents

Abstract

The invention provides a Chinese medicament for treating dyslipidemia and atherosclerosis, and belongs to a Chinese medicament preparation. The invention provides the Chinese medicament which makes a point by deficiency in origin and excess in superficiality, effects a permanent cure by strengthening the spleen and replenishing qi, reduces the generation of lipid turbidity and improves metabolism; and the Chinese medicament also takes temporary solution by expelling phlegm and absorbing clots and inhibits the generation and the development of the atherosclerosis. The Chinese medicament is prepared from the following components by weight: 15 to 25g of gynostemma pentaphyllum, 10 to 15g of tuckahoe, 5 to 15g of pinellia tuber, 15 to 25g of red sage root, 15 to 25g of rhizome of Sichuan lovage, 10 to 15g of grass-leaaved sweetflag, 10 to 20g of curcuma root, 10 to 25g of hawthorn and 15 to 25g of milkvetch root.

Description

A kind of treatment dyslipidemia and atherosclerotic Chinese medicine

Technical field:

The present invention belongs to Chinese medicine preparation.

Background technology:

Atherosclerosis is that the mankind are disabled, first of the lethal important diseases, atherosclerosis is mainly involved aorta, coronary artery, cerebral arteries and renal artery, causes that the organ blood of being supplied for obstacle, causes these organ generation ischemic pathological changes.Can cause diseases such as angina pectoris, myocardial infarction, cerebrovascular accident, brain atrophy, lower limbs necrosis." damage " of arterial endothelium and the toxicity environment of " dyslipidemia " are to impel atherosclerotic one of the main reasons.

Atherosclerotic clinically at present measure mainly comprises:

1. motion: motion can improve the vasodilation function of endothelium-dependent relaxation, increases the gene expression of nitricoxide synthase.

Exercise therapy has certain effect, but curative effect remains further to be investigated.

2. alternative medicine: replenish protection blood vessel endothelium materials such as L-arginine, prostacyclin analogs.

3. the inhibitor or the antagonist of endothelium source property contracting agent: as angiotensin converting enzyme inhibitor, angiotensin ii receptor antagonist, endothelin antagonist, thromboxane A2 synthase inhibitor and thromboxane receptor antagonist etc., can reduce the generation of materials such as angiotensin, Endothelin, thromboxane A2, have certain effect preventing and treating atherosclerosis.These preparations have been taken many side effect for a long time as cough, hemorrhage etc.

4. transfer the fat treatment: hypercholesterolemia is to bring out atherosclerotic key factor, participates in promoting the formation of tunica intima atheromatous plaque.Clinical trial confirms that statins can raise the expression of endothelial nitric oxide synthase, improves endothelial function, stablizes atherosclerotic plaque, reduces vascular lesion.

But the untoward reaction of statins also more and more comes into one's own: (1) myopathy, this is a kind of rhabdomyolysis than serious adverse effects, muscular tissue is destroyed, discharge a large amount of Myoglobin, the blood creatine, phosphokinase (CPK) content obviously raises (be higher than normal value 10 times), and Myoglobin excretes through kidney, and serious situation can cause renal failure even death.(2) to endocrine influence.The synthetic inhibition of cholesterol can reduce the secretion of steroid hormone in the body, has the report simvastatin can cause erection disturbance.(3) gastrointestinal reaction is more common, and that symptom is generally is nauseating, stomachache, diarrhoea, flatulence.(4) hepatic injury, behind the long-term prescription, the transaminase can be 3 times of normal value.In addition, in the use of statins, it should be noted that the interaction of medicine, its untoward reaction significantly increases when share with other drug, and consequence is also relatively more serious.

5. endothelium plantation and gene therapy: research at present will be cultivated after conduit " plantation " is stripping off on the blood vessel of endothelium from the body blood vessel endothelium, be expected to improve this vascular endothelial function, and then play the atherosclerotic effect of control.

But this technology still is in laboratory stage, not in clinical extensive popularization.

More than many reasons seriously hindered atherosclerotic method of many controls and the application of medicine in clinical.And other blood lipid regulation or improve atherosclerotic method of treatment or curative effect very little, or cost an arm and a leg or still in experimentation.Therefore, utilizing the relative features of smaller of natural drug toxic and side effects, therefrom seek the research focus that the medicine that newly improves blood vessel endothelium is just becoming world's medicine and pharmacology, also is one of important content that promotes the Chinese medicine development in science and technology.The development toxic and side effects little, evident in efficacy, have blood lipid regulation, prevent and treat the compound Chinese medicinal preparation of many target spots of atherosclerosis Comprehensive Treatment effect, not only have the market demand widely, also will bring huge social benefit and economic benefit.

Summary of the invention:

The present invention is exactly at the problems referred to above, provides a kind of and sets forth one's views with deficiency in origin and excess in superficiality, to effect a permanent cure, reduces the generation of turbid fat with invigorating the spleen and benefiting QI, improves metabolism; Blood stasis dispelling eliminate the phlegm simultaneously to take stopgap measures, to prevent the treatment dyslipidemia and the atherosclerotic Chinese medicine of atherosclerotic generation development.

For achieving the above object, the present invention adopts following technical scheme, and the present invention prepares by following weight:

Herb Gynostemmae Pentaphylli 15-25g, Poria 10-15g, Rhizoma Pinelliae 5-15g, Radix Salviae Miltiorrhizae 15-25g, Rhizoma Chuanxiong 15-25g, Rhizoma Acori Graminei 10-15g, Radix Curcumae 10-20g, Fructus Crataegi 10-25g, Radix Astragali 15-25g.

Beneficial effect of the present invention:

Be applied to atherosclerosis due to the disorders of lipid metabolism through experiment showed,, have blood lipid regulation, improve insulin resistant, protect vascular endothelial function and prevent and treat arteriosclerotic effect.

The specific embodiment:

Embodiment 1:

The present invention prepares by following weight:

Herb Gynostemmae Pentaphylli 20g, Poria 10g, Rhizoma Pinelliae 10g, Radix Salviae Miltiorrhizae 15g, Rhizoma Chuanxiong 20g, Rhizoma Acori Graminei 10g, Radix Curcumae 15g, Fructus Crataegi 15g, Radix Astragali 25g.

Embodiment 2:

The present invention prepares by following weight:

Herb Gynostemmae Pentaphylli 25g, Poria 15g, Rhizoma Pinelliae 10g, Radix Salviae Miltiorrhizae 20g, Rhizoma Chuanxiong 15g, Rhizoma Acori Graminei 10g, Radix Curcumae 10g, Fructus Crataegi 20g, Radix Astragali 20g.

Embodiment 3:

The present invention prepares by following weight:

Herb Gynostemmae Pentaphylli 15g, Poria 10g, Rhizoma Pinelliae 5g, Radix Salviae Miltiorrhizae 25g, Rhizoma Chuanxiong 25g, Rhizoma Acori Graminei 10g, Radix Curcumae 20g, Fructus Crataegi 25g, Radix Astragali 15g.

This achievement proves that fully the present invention has obvious blood lipid regulation and improves atherosclerotic effect on pathomorphology, biochemistry, molecule and cytobiology level on the basis of early stage a large amount of clinical researches and a series of Basic Experiment Study.

The accent fat effect of being eliminated the phlegm in blood stasis dispelling side in experiment one

1. materials and methods

1.1 animal

Select the Wister rat for use, body weight 400-450g provides lot number NO:0909120,50 by Shen doctor's Experimental Animal Center

1.2 grouping and modeling method

Rat is divided into five groups at random by body weight, sex.

Normal group: first group

Modeling group: second group: the 3rd group of the blank group of model: the 4th group of metformin group: the 5th group of zhibituo group: the blood stasis dispelling side's group of eliminating the phlegm

Method by Reaven is changed slightly, duplicates the IR rat model.Normal group is fed normal diet, wherein sugar, fat and protein account for 50%, 20% and 30% of total calorie card amount, the modeling group is fed high lipid food, the wherein fatty calorie amount that provides is up to 59%, carbohydrate only provides the heat about 20%, the Main Ingredients and Appearance of fat is a lard stearin, is mainly satisfied fatty acid.Two groups of total calorie card amounts are identical.

1.3 medicine and medication

Metformin hydrochloride: northeast the 6th pharmaceutical factory produces.[defending the accurate word (1996) of medicine No. 000897]

Zhibituo: Chengdu Diao 9 Wang pharmaceutical factory production.[river defend the accurate word of medicine (1996) year No. 012750]

The blood stasis dispelling side of eliminating the phlegm: be made up of the Radix Astragali, Poria etc., water boiling concentration is concentrated into 2ml by 7 times of amounts of adult's dosage, and day is once irritated stomach.

Metformin, zhibituo all by 7 times of amounts of adult's dosage, are made the 2ml suspension, and day is once irritated stomach.Altogether; Eliminate the phlegm blood stasis dispelling side through decocting, be concentrated into 2ml by 7 times of amounts of adult's dosage, day is once irritated stomach, around being total to.

The empty group of normal group and mould gives normal saline 2ml, and day is once irritated stomach, around being total to.

2. reagent and method

2.1 serum levels of triglyceride, cholesterol: adopted enzymic colorimetric, test kit: U.S. Beckman company.The serum high-density LP one-step method, test kit: Japanese First Chemical Corp.; U.S. Beckman CX5 type biochemistry analyzer.

2.2 get blood before the experiment, use etherization, the eye socket vein is got blood.Rat is put to death after getting blood with 20% urethane anesthesia (5ml/kg.ip) carotid artery in the experiment back.

3. statistical procedures

Measurement data is checked with t, the date processing pems of vital statistics teaching and research room of Huaxi Medical Univ statistical package.

4. result:

4.1 body weight change after the modeling

Weight increase is increased obviously than normal group (weight increase 11.89g after its modeling) by 32.74-34.08g after the modeling of modeling group rat, illustrates that high lipid food can cause rat body weight and obviously increase, and is to cause fat one of the main reasons.

4.2 respectively organize the variation of blood fat after the medication

The empty group of mould TG is significantly higher than normal group, P＜0.01, the low and normal group of mould empty group HDL, P＜0.05.Mould empty group TC and normal group compare P＞0.05.The blood stasis dispelling side group TG that eliminates the phlegm is lower than the empty group of mould, P＜0.05.Zhibituo also has the reduction effect to the TG of IR rat, but P＞0.05.Eliminate the phlegm the empty group of blood stasis dispelling side, two biguanide, zhibituo, four groups of TC of losartan and mould relatively, P＞0.05.Eliminate the phlegm the empty group of blood stasis dispelling side, metformin, zhibituo, four groups of HDL of losartan and mould relatively, all P＞0.05.

Experiment two: the regulating action of the insulin resistant that causes is damaged by the blood stasis dispelling side of eliminating the phlegm to high fat

1. animal

Select the Wister rat for use, body weight 400-450g provides lot number NO:0909120,50 by Shen doctor's Experimental Animal Center.

2. divide into groups and modeling method

Rat is divided into five groups at random by body weight, sex.

Normal group: first group

Modeling group: second group: the 3rd group of the blank group of model: the 4th group of metformin group: the 5th group of zhibituo group: the blood stasis dispelling side of eliminating the phlegm

3. medicine and medication

Metformin hydrochloride: northeast the 6th pharmaceutical factory produces.[defending the accurate word (1996) of medicine No. 000897]

Zhibituo: Chengdu Diao 9 Wang pharmaceutical factory production.[river defend the accurate word of medicine (1996) year No. 012750]

The blood stasis dispelling side of eliminating the phlegm: be made up of the Radix Astragali, Poria etc., water boiling concentration is concentrated into 2ml by 7 times of amounts of adult's dosage, and day is once irritated stomach.Metformin, zhibituo all by 7 times of amounts of adult's dosage, are made the 2ml suspension, and day is once irritated stomach.Altogether; Eliminate the phlegm blood stasis dispelling side through decocting, be concentrated into 2ml by 7 times of amounts of adult's dosage, day is once irritated stomach, around being total to.The empty group of normal group and mould gives normal saline 2ml, and day is once irritated stomach, around being total to.

4. statistical procedures

Measurement data is checked with t, the date processing pems of vital statistics teaching and research room of Huaxi Medical Univ statistical package.

5. result

Respectively organizing IRI, ISI before and after the medication changes

Group treatment front and back, the blood stasis dispelling side IRI that eliminates the phlegm has significant difference, P＜0.05, and the two croak IRI treatment of diformazan front and back are P＜0.05 relatively, and zhibituo group IRI variation is not obvious, P＞0.05.Eliminate the phlegm blood stasis dispelling side and metformin group relatively, P＞0.05.

Respectively organize ISI before and after the medication and change comparison sheet

ISI significant difference before and after the treatment, P＜0.05, ISI is changed significantly P＜0.05 before and after losartan, two groups of treatments of metformin.Blood stasis dispelling side and the metformin group of eliminating the phlegm be P＞0.05 relatively.Zhibituo group ISI changes not obvious, P＞0.05.

6. conclusion

Blood stasis dispelling side's group treatment bleeding from anus TG that eliminates the phlegm reduces significantly, P＜0.05, IRI significantly reduces before the treatment, P＜0.05, ISI raises significantly than the treatment back, and P＜0.05 points out the blood stasis dispelling of eliminating the phlegm can obviously improve the insulin sensitivity of rat, improve the insulin resistant phenomenon, it is relevant to infer that its mechanism and its have an effect for reducing blood fat.IRI, ISI are changed significantly before and after losartan, the treatment of metformin group, and also there is the IR of improvement effect P＜0.05.

Test three the present invention to the influence of vascular endothelial injury label vWF and the influence of endothelium form

1 experiment material

1.1 experimental cell: Human umbilical vein endothelial cells cell line (HUVECs) is preserved center (ECV--304) available from the typical thing of China.

1.2 medicine and reagent: hyperlipemia is clear, pastille serum; Calf serum: Hangzhou Sijiqing Biological Engineering Material Co., Ltd.'s product; DMEM-1640 dry powder: U.S. GibcoBr1 company product; MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt, trade name tetrazolium bromide) and trypsin: Huamei Bio-Engrg Co.,'s product; Dimethyl sulfoxide etc. are homemade analytical pure, 1.3 key instrument equipment: CO2 incubator: Fisher Scientific; IX50 type inverted phase contrast microscope: OLYMPUS; Superclean bench: Nantong instrument plant; The XB-K-25 cell counting count board; Microplate reader: Labsystems Multiskan Ascent; 722s spectrophotometer: school, Shanghai light instrument plant; MILLI-Q ultrapure water machine: U.S. MILLIRORE.

2. experimental technique

2.1 cell culture

2.1.1 cell recovery: frozen peace bottle in liquid nitrogen is taken out, putting into 37 ℃ of water-baths rapidly makes it melt at 1 minute, open the peace bottle under the aseptic condition, cell suspension is drawn in the 100ml culture bottle, supplies culture fluid 10ml (10% calf serum+90%DMEM), put CO2 incubator (5%CO2,37 ℃) the middle cultivation, treat cell attachment (after about 48 hours), change culture fluid once, to cell grow up to monolayer and be paved with bottle at the bottom of the time can go down to posterity.

2.1.2 passage: inhale and remove culture fluid, add 1ml 0.25% trypsin, rock culture bottle gently, make trypsin cover the whole bottle end.The Cytoplasm retraction is seen in room temperature (more than 25 ℃) digestion 3-5 minute under the mirror, when intercellular substance increases, add 2ml and contain the serum culture fluid, stops digestion.The elbow suction pipe is drawn culture fluid in the bottle, and piping and druming bottle parietal cell is made the individual cells suspension repeatedly, divides in the new culture bottle of packing into after the counting cells quantity, supplies culture fluid and cultivates, and experimental cell is left and taken in circulation so repeatedly.When treating that cell reaches enough experimental applications, the gained cell is made cell suspension, adjusting cell number is 5 * 106, is used for experiment.

2.1.3 pastille serum cell toxicity test (MTT): with the cell of 0.25% trypsinization monolayer culture, be made into the individual cells suspension with the DMEM culture fluid that contains 10% calf serum, the adjustment cell concentration is 1 * 105/ml, is inoculated in 96 well culture plates, every hole 200 μ l cell suspension.Culture plate is moved in the CO2 incubator, under 37 ℃ of 5%CO2 and saturated humidity condition, cultivate after 24 hours, abandon culture fluid, add the DMEM1640 culture fluid respectively and add various drug serum culture fluid, grouping sees Table 1 in detail.(every group 6 hole).

Table 1MTT experiment grouping and serum ratio

Rat blood serum with no pastille is a matched group, continues to cultivate after 24 hours, 48 hours, adds 0.5% MTT (5mg/ml), every hole 20 μ l, and 37 ℃ were continued to hatch 4 hours.Stop cultivating, the careful suction abandoned culture supernatant in the hole, and every hole adds 150 μ lDMSO, and 37 ℃ vibrated 10 minutes, uses microplate reader and detects the 490nm OD of place value.

The influence of 24 hours and the survival of 48 hours drug serum pair cells can be calculated with cell survival rate.Average OD value * 100% that the average OD value that cell survival rate=medication hole is measured/control wells is measured

2.2EC damage model is made and grouping

2.2.1 biochemical indicator: well-grown endotheliocyte, make the individual cells suspension, adjust cell number and be 5 * 106 and be inoculated in 96 orifice plates (Costar company), supply culture fluid to 200 μ l, treat the cell attachment fusion growth, the culture fluid that inclines adds hyperlipemia by different proportion and reaches pastille serum clearly.Cultivated 24 hours and 48 hours in 37 ℃ of 5%CO2 incubators.Every group 6 hole.

24 hours:

Blank group: ECV+ normal mouse serum 20%----24H

Matched group: the clear 10%+ normal rat of ECV+ hyperlipemia serum 10%----24H

Modeling group: the clear 20%----24H of ECV+ hyperlipemia

Chinese drug-treated group (height): the clear 10%+ Chinese medicine of ECV+ hyperlipemia serum (height) 10%----24H

Chinese drug-treated group (often): the clear 10%+ Chinese medicine of ECV+ hyperlipemia serum (often) 10%----24H

Western medicine group: the clear 10%+ Western medicine of ECV+ hyperlipemia serum 10%----24H

48 hours:

Blank group: ECV+ normal mouse serum 20%--24H removes supernatant+normal mouse serum 20%----24H

Matched group: the clear 20%----24H of ECV+ hyperlipemia removes supernatant+normal mouse serum 20%----24H

The modeling group: the clear 20%----24H of ECV+ hyperlipemia removes the clear 20%----24H of supernatant+hyperlipemia

Chinese drug-treated group (height): the clear 20%----24H of ECV+ hyperlipemia removes supernatant+Chinese medicine serum (height) 20%----24H

Chinese drug-treated group (often): the clear 20%----24H of ECV+ hyperlipemia removes supernatant+Chinese medicine serum (often) 20%----24H

The Western medicine group: the clear 20%----24H of ECV+ hyperlipemia removes supernatant+Western medicine serum 20%----24H

All the other culture fluid add serum-free DMEM

2.2.2 morphological index: cell culture: cell suspension is added in the 100ml culture bottle, supply culture fluid to 10ml, surplus method is the same.Hyperlipemia reaches the ratio that applies of pastille serum clearly: the ratio of various rat blood serums and culture fluid (serum-free DMEM) is 20: 80

Compare p＜0.05 with the modeling group; # and matched group be p＜0.05 relatively; Modeling group 24 hours and 48 hours comparison p＜0.05.

The transmission electron microscope form: the normal endothelial cell form: cell membrane is clear, and more mitochondrion is arranged within the cell, and ridge is high-visible; Rough endoplasmic reticulum is clearly visible, has ribosome on it; Also can see more free ribosome in the kytoplasm; Nuclear membrane is clear.

24 hours: (1) was to making group: fine hair reduces on the cell membrane, visible primary lysosome and secondary lysosome, and mitochondrion has vacuolation.(2) modeling group: fine hair reduces on the cell membrane, and the interior mitochondrial crista fracture of Cytoplasm is serious (but can see); Can see secondary lysosome; Reticulum dilatation.(3) Chinese drug-treated group (often): nucleus has not obvious incisura, and rough endoplasmic reticulum is slightly expanded, and has ribosome on it; Mitochondrial crista is obvious, accidental swelling mitochondrion.(4) Chinese drug-treated group (height): more fine hair is arranged on the cell membrane, and mitochondrion is normal, and mitochondrial crista is clear; Endoplasmic reticulum does not have enlargement, adheres to than polysome; Nucleus has incisura.(5) Western medicine group: fine hair reduces on the cell membrane, and endoplasmic reticulum has slight expansion, accidental cavity line-transect plastochondria, and mitochondrial crista is as seen; Cellular morphology has variation.

48 hours: (1) is to making group: fine hair shortened on the cell membrane, and the mitochondrion vacuolation is more obvious, reticulum dilatation, visible primary lysosome.(2) modeling group: the fine hair fracture disappears the complete vacuolation of mitochondrion in the Cytoplasm on the cell membrane; Reticulum dilatation; The nucleus distortion has incisura.(3) Chinese drug-treated group (often): cellular morphology is normal, and nucleus has incisura, and fine hair reduces on the cell membrane, and endoplasmic reticulum has slight expansion; The accidental cavity sample of mitochondrion, mitochondrial crista can be seen.(4) Chinese drug-treated group (height): nucleus is smooth, and mitochondrial crista is clearly visible; Has ribosome on the rough endoplasmic reticulum; Visible free ribosome within the cell; Fine hair reduces on the cell membrane.(5) Western medicine group: fine hair reduces on the cell membrane, accidental mitochondrion cavity, and primary lysosome is rare, reticulum dilatation, nucleus distortion incisura is obvious.

3. conclusion

The present invention damages the influence of EC vWF clearly to hyperlipemia

3.1vWF damage with EC: vWF is by vascular endothelial cell and megalokaryocyte is synthetic and secretion, is a kind of blood plasma, endothelial cell surface and particulate glycoprotein of platelet α of being present in.VWF has two kinds of main biological functions, and a kind of is to mediate hematoblastic adhesion, gathering and thrombosis at the blood vessel injury place; Another kind is to be combined into the VIII factor/vWF complex with VIII factor precursor, makes the VIII factor active become stable.Vascular endothelial cell contains a certain amount of vWF, but when endothelial cell damage, degeneration, when coming off etc., endotheliocyte produces vWF to be increased.Confirmed extensively that in various vascular conditions the increase of vWF level is one of mark of vascular endothelial cell damage.

3.2 the present invention is to the influence of vWF: this experiment is 24 hours result show, with hyperlipemia clear (10%) and three kinds of drug serums (10%) co-cultivation ECV, after 24 hours with set matched group (10% hyperlipemia clear+10% rat blood serum) relatively, the vWF amount is starkly lower than matched group in its culture fluid, (p＜0.05) has statistical significance; Drug serum group of the present invention is better than Western medicine group (p＜0.05), has statistical significance; And the effect of high dose group is better than normal dosage group (p＜0.05), has statistical significance.Show drug serum of the present invention for prevent hyperlipemia clear due to ECV damage, degeneration, cause that vWF increases in its culture fluid, its effect is better than atorvastatin, and high dose group of the present invention is better than normal dosage group.48 hours result shows, behind hyperlipemia clear (20%) damage ECV, impose drug serum (20%) or rat blood serum (matched group) again, its culture fluid vWF detects and shows that the high agent of Chinese medicine, normal agent group and Western medicine group are starkly lower than matched group (p＜0.05), have statistical significance; Drug serum group of the present invention is better than Western medicine group (p＜0.05), has statistical significance; And the effect of high dose group is better than normal dosage group (p＜0.05), has statistical significance.Show for hyperlipemia clear due to the ECV damage, drug serum of the present invention can reduce vWF content in its culture fluid, its effect is better than atorvastatin, and high dose group of the present invention is better than normal dosage group.

Test the influence of four the present invention to the vascular endothelial cell fibrinolytic system of the clear damage of hyperlipemia

1 experiment material

1.1 experimental cell: the same

1.2 medicine and reagent: hyperlipemia is clear, pastille serum: do certainly as preceding method; Calf serum: Hangzhou Sijiqing Biological Engineering Material Co., Ltd.'s product; DMEM-1640 dry powder: U.S. GibcoBr1 company product; MTT (3-(4,5-dimethylthiazole-2)-2,5-diphenyl tetrazole bromine salt, trade name tetrazolium bromide) and trypsin: Huamei Bio-Engrg Co.,'s product; Dimethyl sulfoxide etc. are homemade analytical pure.Human plasma GMP-140 enzyme marking reagent box, t-PA, PAI determination of activity test kit: Shanghai sun biotech company;

1.3 key instrument equipment: CO2 incubator: Fisher Scientific; IX50 type inverted phase contrast microscope: OLYMPUS; Superclean bench: Nantong instrument plant; The XB-K-25 cell counting count board; Microplate reader: LabsystemsMultiskan Ascent; 722s spectrophotometer: school, Shanghai light instrument plant; MILLI-Q ultrapure water machine: U.S. MILLIRORE.

2 experimental techniques are the same

3. index detects:

3.1 specimen is left and taken PAI-1, t-PA and is all got 96 orifice plate culture fluid supernatants

3.2 detection method t-PA, PAI-1 determination of activity: chromophoric substrate method.

4. statistical method: measurement data with average ± standard deviation (x ± s) expression relatively behind the variance test of homogeneity, does the q check of variance analysis between group, with p＜0.05 for statistical significance is arranged, with the SPSS11.0 deal with data.

5. result: t-PA, PAI-1 determination of activity the results are shown in following table

Annotate: * and modeling group be p＜0.05 relatively; # and matched group be p＜0.05 relatively; Modeling group 24 hours and 48 hours comparison p＜0.05.

6. conclusion: the present invention damages the influence of EC t-PA, PAI-1 clearly to hyperlipemia

The present invention is to the influence of t-PA, PAI-1: modeling group t-PA descends, PAI-1 raises as previously mentioned.With hyperlipemia clear (10%) and three kinds of drug serums (10%) co-cultivation ECV, establish matched group (10% hyperlipemia clear+10% rat blood serum) simultaneously, 24 hours result shows, compare with matched group, the active rising of the t-PA that pastille serum is three groups, the active decline of PAI-1 (p＜0.05) all have statistical significance; Drug serum group of the present invention is better than Western medicine group (p＜0.05), has statistical significance; And the effect of high dose group is better than normal dosage group (p＜0.05), has statistical significance.Show drug serum of the present invention for prevent hyperlipemia clear due to ECV t-PA actively descend, PAI-1 is active raises, and then improves the abnormal fibrinolytic activities due to the damage ECV, also has this effect in atorvastatin.48 hours result shows, behind hyperlipemia clear (20%) damage ECV, imposes drug serum (20%) or rat blood serum (matched group) again, and the high agent of Chinese medicine, normal agent group and Western medicine group PAI-1 activity are starkly lower than matched group (p＜0.05), have statistical significance; Matched group t-PA activity is starkly lower than of the present invention group and Western medicine group (p＜0.05), has statistical significance; Show that of the present invention group and Western medicine group can reduce all that ECV t-PA due to the clear damage of hyperlipemia actively descends, PAI-1 is active raises, reconcile the imbalance of t-PA/PAI-1, effect wherein of the present invention is better than atorvastatin, and the effect of high dose group is better than normal dosage group (p＜0.05), has statistical significance.Show for hyperlipemia clear due to the ECV damage, drug serum of the present invention can be by regulating the imbalance of t-PA/PAI-1, and then improve fibrinolytic function, improve hyperlipemia clear due to the ECV damage.Its effect is better than atorvastatin, and high dose group of the present invention is better than normal dosage group.

Test the influence of five the present invention to ECV P-Selectin

1.1 material

1.1.1 animal and cell: the same

1.1.2 medicine and reagent: atorvastatin (lipitor), the 10mg/ sheet, Pfizer Inc. produces.Granule of the present invention.By Hospital Attached to Liaoning Inst. of Traditional Chinese Medicine Drug Manufacturing Room preparation, the anti-people P-selectin of goat monoclonal antibody, the anti-goat IgG of FITC labelling rabbit, Beijing Zhong Shan Bioisystech Co., Ltd.: the pcr amplification primer: according to document [4] data design primer, primer sequence: upstream: 5 ,-CAA TCA CTT TTA GCC AAATAC C-3; The downstream: 5 ,-TTA TAA TTC AGC AAC CTG AAC TG-3.With beta-actin (β-actin) does confidential reference items: β-actin upstream: 5 ,-AAG GAT TCC TAT GTG GGC-3; The downstream: 5 ,-CAT CTC TTG CTC GAA GTC-3,, synthetic by the biological Dalian of treasured company limited, the explanation dilution is pressed in 20 ℃ of preservations before using.Trizol：GiBCo。Chloroform: Shenyang Sheng Wei chemical reagent work.Isopropyl alcohol, dehydrated alcohol: Shenyang reagent one factory.RT-PCR test kit: precious biological Dalian company limited, lot number: 10815

1.1.3 key instrument equipment: CO2 incubator, PCR instrument: German Whatman.Biometra; BIOFUGE28S low-temperature and high-speed centrifuge: German HZRAZUS; DY89-I type electric driven glass refiner: Ningbo Xin Zhike device institute; The ultrasonic cell pulverization machine of JY92-II: Ningbo Xin Zhike device institute; UV-300 type ultraviolet spectrophotometer: Britain; MILLI-Q ultrapure water machine: U.S. MILLIRORE; DYY-III5 type voltage stabilization and current stabilization electrophresis apparatus: Liuyi Instruments Plant, Beijing; Superclean bench: Nantong instrument plant; MDF382E ultra cold storage freezer: Japanese SANY; SBH circulation ability of swimming vacuum pump: Great Wall, Zhengzhou.H.HWZI.600 type electric heating constant temperature water bath: Huanghua, Hebei province space flight instrument plant; Chemi Imager 5500 gel images acquisition system: AlphaInnotech.

1.1.4 index detects

Adopt double-antibody sandwich insoluble enzyme immunity test method

The PT-PCR method detects EC Ps mRNA

Western-Bloting detects endotheliocyte P-Selectin protein expression

1.2 experimental technique

1.2.1 cell culture: well-grown endotheliocyte, use 0.25% trypsinization, be inoculated in the Tissue Culture Plate of 96 orifice plates with 5 * 106 cells/well, treat the cell fusion growth, add culture fluid and apply factor, 37 ℃ of 5%CO2 are hatched, and are respectively action time 24 hours, 48 hours, detect P-Selectin in the culture fluid supernatant respectively.

1.2.2 the ratio of experiment grouping and various serum: same vWF.

1.2.3 specimen is left and taken: get 96 hole this culture fluid supernatants.

1.3 detection method and step

1.3.1 method: adopt double-antibody sandwich insoluble enzyme immunity test method.

1.3.2PT-PCR method detects EC Ps mRNA

1. the extraction of cell total rna: get cultured cell, cell concentration is about 5 * 106, scrapes cell with scoop, and is centrifugal, clean.Add 1ml Trizol in centrifuge tube, concussion adds the 0.2ml chloroform, shook 15 seconds, centrifugal 15 minutes of 4 ℃/2000rpm gets supernatant 0.5ml, add isopropyl alcohol 0.5ml, concussion ,-20 ℃ of anti-putting 15 minutes, 2000rpm is centrifugal 15 minutes under 4 ℃ of conditions, abandon supernatant, precipitation adds 75% ethanol 1ml, centrifugal 15 minutes of 12000rpm, abandon supernatant, drying at room temperature 20 minutes.Add in the tri-distilled water of 10 μ l DEPC processing.

2. extract the RNA purity detecting: get 1 μ l, add DEPC and handle tri-distilled water 79 μ l, ultraviolet spectrophotometer detects 260nm, the OD of 280nm place value respectively, with calculating RNA purity and concentration (RNA purity=OD260/OD280).The purity of surveying all at 1.76-1.97.

3. the total RNA reverse transcription by cell extraction is cDNA: form inverse transcription reaction liquid (totally 20 μ l) by following condition:

MgCl2 4μl

10×RNA?PCR?Buffer 2μl

RNase?Free?dH2O 8.5μl

DNTP Mixture (each 10mM) 2 μ l

RNase?Inhibitor 0.5μl

AMV?Reverse?Transcriptase*1 1μl

Random?9?mers 1μl

Laboratory sample RNA 1 μ l

Carry out reverse transcription reaction by following condition: 30 ℃ → 10 minutes, 45 ℃ → 30 minutes, 99 ℃ → minute, 5 ℃ → 5 minutes.

4. PCR reaction: pcr amplification P-Selectin segment (210bp), do confidential reference items with-actin (532bp).Prepare PCR reactant liquor (totally 80 μ l) by following composition:

MgCl2 6μl

10×RNA?PCR?Buffer 8μl

Sterile purified water 63.5 μ l

TaKaRa?Taq 0.5μl

Upstream specific PCR primer 1 μ l

Downstream specific PCR primer 1 μ l

Reaction condition: 94 ℃ of pre-degeneration began circulation in 2 minutes, 94 ℃ 30 seconds → 52 ℃ 30 seconds → 72 ℃ 1 minute, 30Cycles altogether.Last 72 ℃ were fully extended 10 minutes.Every pair of primer done a PCR reaction.

5. electrophoresis and imaging: product 2% sepharose electrophoresis (150v 30min) of putting together away, the own ingot of bromine soaked 10 minutes, and the imaging of gel instrument is also analyzed.More than react triplicate, reach an agreement.

1.3.3Western-Bloting detect endotheliocyte P-Selectin protein expression

1. cracking memebrane protein: the cell of leaving and taking is put in the 1.5mlEP pipe, adds 400 μ l protein lysates.Ultrasonication is 20 seconds in 0 ℃ of frozen water, and 20 seconds at interval, 5 times repeatedly.4 ℃ of standing over night.In 4 ℃/12000rpm centrifugal 1 hour, get supernatant and be put in the 1.5mlEP pipe.

2. decide protein concentration (phenol reagent process): sample thief 50 μ l, standard protein (with the BSA of the 1mg/ml of PBS preparation) and each 50 μ l of blank water.More than three groups all add alkaline copper liquid 2.5ml, placed 20 minutes, added the 0.25ml phenol reagent 30 minutes.With the blank well zeroing, measure 650nm wavelength place optical density value on the spectrophotometer.

The protein concentration of surveying=mensuration pipe OD puts/standard protein OD value * 1 (mg/ml).

3. polyacrylamide gel electrophoresis (SDS-PAGE): irritate separation gel during encapsulating earlier, about 20 minutes gelling to be separated solid after, irritate again and concentrate glue, add comb, extract comb after the polymerization.In well, add and respectively organize sample 50 μ g.The wet type electrophoresis tank adds electrophoresis liquid, 120V, 1 hour.

4. transfer printing: (1) cuts 6 3MM filter paper and a nitrocellulose filter, and what its size should be with glue is big or small identical.The 3MM filter paper and the nitrocellulose filter that shear were soaked in transfering buffering liquid 3-5 minute.(2) transfer device successively is installed: the first day of the lunar month is expected that support lies in the pallet that contains transfering buffering liquid, on the first day of the lunar month material support, put a sponge.3 3MM filter paper alignment are placed on the sponge anti-successively cellulose membrane, gel, other 3 3MM filter paper and the sponge put.When placing each layer, all to remove the bubble between them.Clamp above-mentioned each layer with north material support at last, put into electrotransfer groove (cellulose membrane one side is by anodal, and glue one side is by negative pole).(3) energized, voltage 50V, electric current 100mA, transferase 12 hour.(4) after the transfer end, remove north material support, remove each layer successively, use TBS (Tris Buffer Nacl) to wash for 5 seconds cellulose membrane then.

5. sealing: nitrocellulose filter is placed in the plate, adds confining liquid (its amount had been soaked film and got final product).Vibrated gently under the room temperature 1-2 hour, and took out film and wash 5 minutes * 2 times with TBS.

6. with dilution in 1: 300,4 ℃ of overnight incubation .TTBS washed 5 minutes * 2 times with the T-Tween-20 (TTBS) that contains 1%BSA for reaction of target protein and first antibody and second antibody reaction: P-Selectin.The anti-goat IgG of HRP labelling rabbit, with dilution in 1: 2000, incubated at room 2 hours was taken out back TTBS washing 5 minutes * 2, again with TBS washing 5 minutes.

7. colour developing and analysis: film was put in luminescence reagent 1 minute, and exposure is 1 minute in the magazine, develops photographic fixing 5 minutes 5 minutes.Gel images acquisition system analysis result.

1.4 statistical procedures

All data are represented with X ± s, relatively adopt variance analysis between group, use the SPsS10.0 software processes.

1.5 result

The present invention sees the following form to the influence of EC solubility P-Selectin

Annotate: * and modeling group be p＜0.05 relatively; # and matched group be p＜0.05 relatively; △ modeling group 24 hours and 48 hours comparison p＜0.05.

The present invention sees the following form to the influence (FCM) of EC surface P-Selectin

Annotate: * and modeling group be p＜0.05 relatively; # and matched group be p＜0.05 relatively; Modeling group 6 hours and 12 hours comparison p＜0.05.

1.6. conclusion

1. the present invention is to the influence of solubility Ps: the mRNA of Ps molecule exists different splicing forms in megalokaryocyte and the EC, wherein a kind of cytoplasmic domain mRNA encoded protein matter that lacks is secreted in the blood, it is the main source of solubility Ps in the blood plasma (soluble P-selectin sPs), the Ps of EC surface expression also can come off and enter blood simultaneously, becomes sPs.Structurally, sPs generally lacks its corresponding film and wears film and endochylema part in conjunction with adhesion molecule, its molecular weight is little than corresponding membrane in conjunction with Ps also, sPs have usually Ps in conjunction with active, therefore may play a role as the approach that body is regulated the cell adhesion effect.This experimental applications ELISA method detects the Ps of ECV culture supernatant, is equivalent to intravital sPs and changes.Experimental result shows that clear its modeling group of the ECV Ps that cultivates of hyperlipemia has statistical significance apparently higher than matched group and treatment group (p＜0.05), shows that hyperlipemia can cause ECV sPs clearly and increase; Cultivation results showed in 24 hours, and drug serum group sPs of the present invention is starkly lower than modeling group, matched group and Western medicine group, and high dose group is better than normal dosage group, show drug serum of the present invention have the hyperlipemia of preventing clear due to Ps express, and be dose dependent.Atorvastatin group Ps is starkly lower than modeling group, matched group, show the atorvastatin drug serum also have the hyperlipemia of preventing clear due to sPs express.Cultivation results showed in 48 hours, and modeling group Ps has statistical significance apparently higher than 24 hours groups (p＜0.05), showed that the prolongation of hyperlipemia clear action time is expressed influential to Ps.Drug serum group sPs of the present invention is starkly lower than modeling group, matched group and Western medicine group (p＜0.05), all has statistical significance, and high dose group is better than normal dosage group, show drug serum of the present invention have reduce hyperlipemia clear due to the sPs expression, and be dose dependent.Atorvastatin group Ps is starkly lower than modeling group, matched group (p＜0.05), all has statistical significance, show the atorvastatin drug serum also have reduce hyperlipemia clear due to the Ps expression.

2. the present invention expresses influence to EC surface Ps: flow cytometer (Flow Cytometer) is collector injection, laser, air technology, gamma-radiation power spectrum art, and technology and the married instrument of fiber fluorescent photometer such as computer, can detect the active substance expression on individual cells surface.The fluorescence two of this experimental applications FITC labelling resists and ECV surface Ps antibodies, the height that the surperficial Ps of fluorescence intensity explanation ECV that detects by FCM expresses.Experimental result shows, with the clear endothelial cells cultured of same concentrations hyperlipemia, modeling group Ps expresses apparently higher than matched group (p＜0.05) when cultivating 6 hours, cultivating 12 hours Ps expresses also apparently higher than matched group (p＜0.05), and 6 hours Ps expresses apparently higher than 12 hours (p＜0.05), all has statistical significance.Ps normally is stored in static platelet α granule and the endotheliocyte weibel-palade corpusculum, normal endothelial cell and platelet surface do not have the Ps expression or are lasting low expression status, after by stimulations such as inflammatory mediator or endothelial cell damage, along with taking off the particle release effect, membrana granulosa and cell membrane melt membrane interaction, causing Ps to insert to cell surface fast expresses, be expressed in cell surface Ps and can discern mediation mutually by rear and lysosome in its endochylema again and move up into the lysosome degraded in the Ps, minority lacks the Ps that strides rear (or this section is degraded) in film section and the endochylema and then gets back in Weibel-Palade corpusculum or the α-granule.Express with the ECV Ps of this experiment and to be higher than 12 hours result in 6 hours, may with move up into cell or be shed in the culture fluid relevant in Ps is behind the ECV surface expression.

The fluorescence intensity that its endotheliocyte Ps that cultivates 6 hours and 12 hours of drug serum group of the present invention expresses all is starkly lower than modeling group, matched group and Western medicine group (p＜0.05), all have statistical significance, show that drug serum of the present invention has and prevent the Ps expression of hyperlipemia due to clear; Atorvastatin group Ps is starkly lower than modeling group, matched group (p＜0.05), all has statistical significance, show the atorvastatin drug serum also have the hyperlipemia of preventing clear due to the Ps expression.This may can be avoided melting film with cell membrane by stabilised blood platelet membrana granulosa with the present invention, stablizes endotheliocyte, stops Ps to insert to cell surface and expresses relevant.

3. the present invention is to the influence of EC Ps mRNA expression

Hyperlipemia with 10% and 20% is cultivated ECV clearly, shows from 24 hours results, and modeling group Ps mRNA expresses apparently higher than blank group (p＜0.05), has statistical significance, shows that hyperlipemia compares with normal rat serum clearly, and ECV Ps mRNA is expressed to be increased; And bestow the clear modeling group of 20% hyperlipemia, its Ps mRNA expresses apparently higher than bestowing the clear matched group of 10% hyperlipemia (p＜0.05), has statistical significance, shows within the specific limits, along with the increase of the clear content of hyperlipemia, the expression of ECV Ps mRNA is also increased; Show from 48 hours results, modeling group Ps mRNA expresses apparently higher than blank group and matched group (p＜0.05), all has statistical significance, and compare with 24 hours modeling group results, Ps mRNA expressed apparently higher than 24 hours (p＜0.05) in 48 hours, have statistical significance, the hyperlipemia that shows same concentrations is time dependence to the Ps mRNA high expressed due to the endothelial cell damage clearly.

The clear co-cultivation ECV of drug serum of the present invention and hyperlipemia, its Ps mRNA expresses and is starkly lower than matched group and Western medicine group (p＜0.05) after 24 hours, all has statistical significance, and high dose group is better than normal dosage group, shows that drug serum of the present invention can prevent that hyperlipemia is clearly to the damage of ECV and then the Ps mRNA high expressed that causes at the DNA transcriptional level.Atorvastatin group Ps mRNA expresses and is starkly lower than matched group (p＜0.05), has statistical significance, shows that the atorvastatin drug serum also can prevent that hyperlipemia is clearly to the damage of ECV and then the Ps mRNA high expressed that causes at the DNA transcriptional level.The group result showed in 48 hours, cultivate through the clear ECV that damages of hyperlipemia with drug serum of the present invention, its Ps mRNA expresses and is starkly lower than matched group and Western medicine group (p＜0.05), all has statistical significance, and high dose group is better than normal dosage group, show that drug serum of the present invention can improve the Ps mRNA high expressed due to the clear damage of hyperlipemia, both alleviates hyperlipemia from the DNA transcriptional level and clearly the p-of damage ECV is selected plain high expressed.Atorvastatin group Ps mRNA expresses and to be starkly lower than matched group (p＜0.05), has statistical significance, shows that the atorvastatin drug serum also can have at the DNA transcriptional level to alleviate hyperlipemia clearly to the Ps high expressed of damage ECV.

Have experiment to show, after endotheliocyte was exposed to n-LDL 24h, Ps mRNA expresses than matched group to be increased, and conformed to this experimental result, and the mechanism that hyperlipidemia causes Ps mRNA to express increase awaits further experimentation.

4. the present invention influences the Ps protein expression

The western protein quantification analysis result of Ps shows, the modeling group p-of 24 hours and 48 hours selects fibroin apparently higher than blank group and matched group (p＜0.05), all has statistical significance, and the Ps protein expression was apparently higher than 24 hours (p＜0.05) in 48 hours, has statistical significance, show in certain concentration range, the concentration that hyperlipemia is clear increases (from 10% → 20%), the Ps protein expression is increased, and the hyperlipemia that shows same concentrations is time dependence to the high expressed of the Ps protein expression due to the ECV damage clearly; The clear co-cultivation ECV of drug serum of the present invention and hyperlipemia, its Ps protein expression expression is starkly lower than matched group and Western medicine group (p＜0.05) after 24 hours, all has statistical significance, and high dose group is better than normal dosage group, shows that drug serum of the present invention can prevent that hyperlipemia is clearly to the damage of ECV and then the Ps albumen high expressed that causes.Atorvastatin group Ps protein expression is expressed and is starkly lower than matched group (p＜0.05), has statistical significance, shows that the atorvastatin drug serum can prevent that also hyperlipemia is clearly to the damage of ECV and then the Ps protein expression high expressed that causes.Cultivate through the clear ECV that damages of hyperlipemia with drug serum of the present invention, its Ps protein expression is expressed and is starkly lower than matched group and Western medicine group (p＜0.05), all has statistical significance, and high dose group is better than normal dosage group, shows that drug serum of the present invention can improve the high expressed of the Ps protein expression due to the clear damage of hyperlipemia.Atorvastatin group Ps protein expression is starkly lower than matched group (p＜0.05), has statistical significance, shows that the atorvastatin drug serum also can alleviate hyperlipemia clearly to damaging the Ps albumen high expressed of ECV.

Test the influence of six the present invention to ECV E-Selectin

1 experiment material

1.1 experimental cell: the same

1.2 medicine and reagent:, the anti-people E-selectin of goat monoclonal antibody, the anti-goat IgG of FITC labelling rabbit, Beijing Zhong Shan Bioisystech Co., Ltd.: the pcr amplification primer: according to document [4] data design primer, primer sequence: upstream: 5 ,-CAA TCA CTT TTA GCC AAA TAC C-3; The downstream: 5 ,-TTA TAATTC AGC AAC CTG AAC TG-3.With beta-actin (β-actin) does confidential reference items: β-actin upstream: 5 ,-AAG GAT TCC TAT GTG GGC-3; The downstream: 5 ,-CAT CTC TTG CTC GAA GTC-3 is synthetic by the biological Dalian of treasured company limited, and the explanation dilution is pressed in 20 ℃ of preservations before using.Trizol：GiBCo。Chloroform: Shenyang Sheng Wei chemical reagent work.Isopropyl alcohol, dehydrated alcohol: Shenyang reagent one factory.RT-PCR test kit: precious biological Dalian company limited, lot number: 10815

2 experimental techniques

2.1 cell culture: method is the same

2.2 passage: method is the same

2.3EC damage model is made and grouping: method is the same

2.4 index detects:

The PT-PCR method detects EC Es mRNA

Western-Bloting detects endotheliocyte E-Selectin protein expression

3. statistical method: measurement data with average ± standard deviation (x ± s) expression relatively behind the variance test of homogeneity, does the q check of variance analysis between group, with p＜0.05 for statistical significance is arranged, with the SPSS11.0 deal with data.

4. result

4.1 the present invention is to EC E-Selectin mRNA influence (RT-PCR)

Modeling group and matched group compare vWF, TXB2 rising in its culture fluid, PGI2 reduces, E-Selectin mRNA expresses, the E-Selectin protein expression increases, the swelling of common light microscopic visible cell film, profile is unclear, and tangible dark particles is arranged in the cell, see mitochondrial swelling under the transmission electron microscope, ridge disappears, reticulum dilatation, and nucleus incisura equivalent damage changes; The present invention and atorvastatin drug serum group and modeling group are relatively, vWF, TXB2 reduce in its culture fluid, PGI2 raises, and EC surface E-Selectin mRNA expresses, the E-Selectin protein expression descends, and the damaging change of visible EC takes a turn for the better under common light microscopic and the transmission electron microscope; Of the present invention group is better than atorvastatin, and its high dose group is better than normal dosage group

5. conclusion:

1. the present invention has the control hyperlipemia clearly to the damage of EC, improves the EC form, the effect of protection EC function.

2. the present invention protects the mechanism of EC function and its reduction EC vWF, TXB2 content, improves PGI2 content, adjusts TXB2, PGI2 balance; 3. the present invention has the effect of preventing EC E-Selectin to express.4. the present invention protects EC, reduces E-and selects one of plain main mechanism of action that act as its control AS of expressing.

Claims (4)

1, a kind of treatment dyslipidemia and atherosclerotic Chinese medicine is characterized in that by following weight preparation:

Herb Gynostemmae Pentaphylli 15-25g, Poria 10-15g, Rhizoma Pinelliae 5-15g, Radix Salviae Miltiorrhizae 15-25g, Rhizoma Chuanxiong 15-25g, Rhizoma Acori Graminei 10-15g, Radix Curcumae 10-20g, Fructus Crataegi 10-25g, Radix Astragali 15-25g.

2, a kind of treatment dyslipidemia according to claim 1 and atherosclerotic Chinese medicine is characterized in that by following weight preparation:

Herb Gynostemmae Pentaphylli 20g, Poria 10g, Rhizoma Pinelliae 10g, Radix Salviae Miltiorrhizae 15g, Rhizoma Chuanxiong 20g, Rhizoma Acori Graminei 10g, Radix Curcumae 15g, Fructus Crataegi 15g, Radix Astragali 25g.

3, a kind of treatment dyslipidemia according to claim 1 and atherosclerotic Chinese medicine is characterized in that by following weight preparation:

Herb Gynostemmae Pentaphylli 25g, Poria 15g, Rhizoma Pinelliae 10g, Radix Salviae Miltiorrhizae 20g, Rhizoma Chuanxiong 15g, Rhizoma Acori Graminei 10g, Radix Curcumae 10g, Fructus Crataegi 20g, Radix Astragali 20g.

4, a kind of treatment dyslipidemia according to claim 1 and atherosclerotic Chinese medicine is characterized in that by following weight preparation:

Herb Gynostemmae Pentaphylli 15g, Poria 10g, Rhizoma Pinelliae 5g, Radix Salviae Miltiorrhizae 25g, Rhizoma Chuanxiong 25g, Rhizoma Acori Graminei 10g, Radix Curcumae 20g, Fructus Crataegi 25g, Radix Astragali 15g.