CN108186877A - For the Chinese medicine composition of syndrome of blood stasis due to qi deficiency myocardial infarction secondary prevention - Google Patents

Abstract

The invention discloses a traditional Chinese medicine composition for the secondary prevention of myocardial infarction due to qi deficiency and blood stasis syndrome. 60 parts and 3 to 10 parts of leeches. The components of the traditional Chinese medicine composition of the present invention act synergistically, and all the drugs are used in combination in a prescribed proportion to achieve the effects of nourishing qi, activating blood, nourishing the heart and protecting the arteries. Arrhythmia, qi deficiency and blood stasis heart failure, etc. have significant therapeutic and preventive effects, and are suitable for secondary prevention of myocardial infarction, especially for the prognosis of patients with both qi and yin deficiency and blood stasis syndrome myocardial infarction. The traditional Chinese medicine composition of the invention has high safety, little side effects and is suitable for long-term administration.

Description

Traditional Chinese Medicine Composition for Secondary Prevention of Myocardial Infarction with Qi Deficiency and Blood Stasis Syndrome

technical field

The invention belongs to the technical field of traditional Chinese medicine, and in particular relates to a traditional Chinese medicine composition for secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome.

Background technique

Cardiovascular disease is currently the leading cause of death in the world. In recent years, the morbidity and mortality of myocardial infarction among urban and rural residents in my country are still on the rise, and tend to be younger. The persistence of various risk factors related to myocardial infarction is the key to the high mortality and disability rates after myocardial infarction. Therefore, it is imminent to actively do a good job in secondary prevention. Secondary prevention of myocardial infarction refers to preventing re-infarction and sudden death after myocardial infarction, and improving the quality of life of patients. The measures mainly include non-drug treatment, surgical treatment and drug treatment. Drug treatment includes antiplatelet agents, statins, β2 receptors Drugs such as body blockers and angiotensin-converting enzyme inhibitors (ACEIs) are used.

In recent years, many clinical evidences have shown that although the use of drugs and reperfusion therapy can significantly reduce the mortality rate of myocardial infarction, there are still many survivors who are disabled or dead due to secondary reinfarction, severe arrhythmia, heart failure and other cardiovascular events . Western medicine put forward the concept of "polypill" and launched a series of compound western medicine preparations as secondary prevention of myocardial infarction, and tried to reduce the occurrence of serious cardiovascular events by reducing blood lipids and stabilizing atherosclerotic plaques. However, the combined application of multiple drugs virtually increases the incidence of adverse drug reactions, and may not necessarily improve the occurrence of myocardial infarction complications.

In the treatment of cardiovascular diseases, traditional Chinese medicine has accumulated rich experience and has clear curative effects. The mechanism involves expansion of coronary arteries, improvement of myocardial ischemia, reduction of myocardial oxygen consumption, improvement of vascular endothelial function, and regulation of lipid metabolism. , multi-level, multi-target effects, to achieve a comprehensive therapeutic effect of overall regulation. Moreover, the traditional Chinese medicine preparation has mild efficacy, and the compatibility of the medicines reduces the toxicity and enhances the efficacy, and the side effects are small, so it is suitable for long-term use as a secondary preventive medicine for myocardial infarction.

At present, in the field of Chinese patent medicine treatment of coronary heart disease, simple "activating blood circulation and removing blood stasis" is the main method. It is a syndrome of deficiency in origin and excess in superficiality. The deficiency in origin is mainly due to qi deficiency, and the excess in superficiality is mainly caused by blood stasis. It is mostly seen in middle-aged and elderly patients. There is a danger of breaking blood and activating blood, consuming righteous energy, aggravating the deficiency at the root, and making the disease more lingering and difficult to heal.

Qishen Yiqi Dropping Pills (Guoyao Z20030139) is made from astragalus, salvia miltiorrhiza, notoginseng, and Jiangxiang. However, since there is no Yin-nourishing traditional Chinese medicine in the prescription of Qishen Yiqi Dropping Pills, it has been found in practical application that after long-term use of Qishen Yiqi Dropping Pills, some patients have symptoms of dryness and heat such as upset and poor sleep. For patients, use astragalus-type warm-dryness traditional Chinese medicine, and take it for a long time to help heat, which is easy to hurt the yin and move the blood. Therefore, Qishen Yiqi Dropping Pills are not suitable for long-term use by patients with deficiency of both Qi and Yin.

The in-house preparation of Guangdong Provincial Hospital of Traditional Chinese Medicine, Tongguan Capsules, is made of astragalus, salvia miltiorrhiza, leech and borneol. After percutaneous coronary intervention (PCI) in patients with acute myocardial infarction of blood stasis type, Tongguan Capsules added to the basic treatment can reduce cardiac enlargement and improve cardiac diastolic and systolic function, and the effect is more obvious as time goes on. However, since the borneol in Tongguan Capsules is a synthetic borneol, which is industrially synthesized by chemical methods from extracts such as camphor and turpentine, it is pungent in nature and slightly cold in taste, and is easy to consume yang energy. Nausea, vomiting, abdominal pain, Symptoms such as urticaria, so it is not suitable for long-term use.

Chinese patent CN101647856B discloses a pharmaceutical composition whose active ingredients are made of astragalus, salvia miltiorrhiza, and leech, and discloses the curative effect of the pharmaceutical composition for treating coronary heart disease, and the pharmaceutical composition is used for qi deficiency and blood stasis syndrome myocardial infarction II The effect of secondary prevention has not been reported. In addition, anti-heart failure and anti-arrhythmia are the key factors for the secondary prevention of myocardial infarction. The improvement effect of ventricular arrhythmia in the recovery period after infarction has not been reported. However, the medicinal composition also uses astragalus-type warm-dryness traditional Chinese medicine, and there is no Yin-nourishing traditional Chinese medicine in the prescription. It can be predicted that long-term use of the prescription will cause patients to be prone to dry-heat symptoms such as upset and poor sleep.

Therefore, in the field of secondary prevention of myocardial infarction, it is still urgent to develop a new generation of traditional Chinese medicine that treats both symptoms and root causes.

Contents of the invention

The object of the present invention is to provide a traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome, so as to solve at least one of the above technical problems.

Another object of the present invention is to provide a preparation method of the above-mentioned traditional Chinese medicine composition to solve at least one of the above-mentioned technical problems.

Another object of the present invention is to provide the application of the above-mentioned traditional Chinese medicine composition to solve at least one of the above-mentioned technical problems.

According to one aspect of the present invention, there is provided a traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome, the active ingredients of which are made of the following raw materials by weight: 5-60 parts of astragalus, 5-60 parts of salvia miltiorrhiza 5-60 parts of Ophiopogon japonicus and 3-10 parts of leeches.

Preferably, the active ingredients of the traditional Chinese medicine composition of the present invention can be prepared from the following raw materials in parts by weight: 10-50 parts of astragalus, 10-30 parts of salvia miltiorrhiza, 10-50 parts of Ophiopogon japonicus and 3-10 parts of leech.

Preferably, the active ingredients of the traditional Chinese medicine composition of the present invention can be prepared from the following crude drugs in parts by weight: 25 parts of astragalus, 15 parts of salvia miltiorrhiza, 25 parts of Radix Ophiopogon japonicus and 5 parts of leeches.

Academician Chen Keji, a master of traditional Chinese medicine, believes that blood stasis syndrome is the basic pathogenesis of coronary heart disease and a basic syndrome throughout the development of coronary heart disease. Professor Deng Tietao, a master of traditional Chinese medicine, believes that internal causes are the main cause of the disease Reason, the basic pathogenesis of coronary heart disease is " deficiency in origin and excess in superficiality ", and deficiency in origin is the important cause of disease of coronary heart disease. Under the guidance of the academic theoretical system of the above two masters of traditional Chinese medicine, the inventor put forward the theory that "the secondary prevention of myocardial infarction should focus on strengthening the body and eliminating pathogenic factors, nourishing qi, promoting blood circulation and resolving phlegm" after a large number of researches and experimental verifications. The invention of the traditional Chinese medicine composition is formulated under the guidance of this theory. In the prescription of the traditional Chinese medicine composition of the present invention, Radix Astragali is the dried root of Astragalus membranaceus (Fisch.) Bge. or Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Bge.) Hsiao of leguminous plant, and returns to the spleen channel, It is sweet, warm and good at invigorating the middle qi, replenishing qi to help the blood flow, and is the monarch drug; Salvia miltiorrhiza is the dry root and rhizome of the labiatae plant Salvia miltiorrhizBge. The main medicine for heart and chest pain is a ministerial drug; the same as the ministerial drug, Ophiopogom japonicas (L.f) Ker-Gawl is the dried root of Liliaceae plant Ophiopogom japonicas (L.f) Ker-Gawl, sweet in taste, slightly bitter, slightly cold in nature, and returns to the stomach, lungs, The Heart Sutra not only has the function of anti-assist, but also has the effect of nourishing yin and promoting body fluid. It reduces the warmness and dryness of Astragalus membranaceus, nourishes the blood and strengthens the body, and avoids the additional effect of breaking the blood and hurting the body; leeches are the leeches Whitmaniapigra Whitman or The dried whole body of Whitmania acranulata Whitman has a salty and bitter taste, enters the liver meridian and blood, and is good at breaking blood, removing blood stasis and dredging collaterals. The above-mentioned medicines are used in combination in the specified ratio, and they have the effects of nourishing qi, activating blood circulation, nourishing the heart and protecting the arteries. It has remarkable therapeutic and preventive effects, and is suitable for secondary prevention of myocardial infarction, especially for the prognosis of patients with myocardial infarction with Qi and Yin deficiency and blood stasis syndrome.

The traditional Chinese medicine composition of the present invention does not contain borneol, and Radix Ophiopogon japonicus is added as a ministerial drug. Ophiopogon japonicus not only contains natural borneol that can replace part of the artificial borneol, but also extracts of Ophiopogon japonicus saponins can strengthen myocardial contraction and increase cardiac output. And anti-arrhythmia effect. In addition, the combination of Ophiopogon japonicus and Astragalus membranaceus reduces the warming and dryness of Astragalus membranaceus, which not only nourishes the blood and strengthens the body, but also avoids the extra effect of breaking the blood and hurting the body. Therefore, the pharmaceutical composition of the present invention is suitable for long-term administration by patients with myocardial infarction. In addition, during the experiment, the inventors found that adding Ophiopogon japonicus can significantly increase the content of calycosin-7-O-β-D-glucoside, rosmarinic acid, formononetin and astragaloside, etc. in the composition. The content of effective compounds, there is a synergistic effect between Ophiopogon japonicus and other components of the Chinese medicine composition of the present invention.

In some embodiments, the traditional Chinese medicine composition of the present invention can be tablets, capsules, powders, oral liquids, pills, ointments, pills, granules or granules.

The traditional Chinese medicine composition used for the secondary prevention of myocardial infarction of qi deficiency and blood stasis syndrome can be taken orally, and the dose is generally 5-30 g of raw medicinal materials per day, and the specific dose can be determined according to the patient's condition and age.

In the acute toxicity test on mice, the maximum tolerated dose (MTD) of the Chinese medicine composition of the present invention is equivalent to 1001 times the clinical intended dose in kilograms of body weight, which is far higher than the maximum 600 times of the pharmaceutical composition described in patent CN101647856B. The tolerated dose shows that the traditional Chinese medicine composition of the present invention has higher safety and is suitable for long-term administration.

According to another aspect of the present invention, the present invention also provides the preparation method of above-mentioned Chinese medicine composition:

Preparation method 1:

This preparation method comprises the steps:

(1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder;

(2) Weigh astragalus, salvia miltiorrhiza and Ophiopogon japonicus according to the proportion, add 4 to 20 times the total weight of the three raw materials to extract 1 to 3 times, each time for 0.5 to 3 hours, combine the extracts, filter (or centrifuge ), the filtrate (or supernatant) is concentrated under reduced pressure into a thick extract with a water content of 15 to 20% or a dry extract with a water content of 3 to 8%, mixed with the leech fine powder described in step (1) Homogenized, adding pharmaceutically acceptable carriers to make various pharmaceutical preparations.

Or further, the filtrate (or supernatant) obtained by extracting Astragalus, Danshen and Ophiopogon japonicus with water is concentrated under reduced pressure to 1/12-1/6 of the volume of the original filtrate to obtain a concentrated solution, which is added with ethanol to the alcohol content 30% to 90%, set aside to settle, filter (or centrifuge), filtrate (or supernatant) is decompressed to recover ethanol, concentrated into a thick extract with a water content of 15 to 20% or made into a water content of 3 to 8 % dry extract, mixed with the leech fine powder described in step (1), adding pharmaceutically acceptable carriers to make various pharmaceutical preparations.

Preparation method 2:

This preparation method comprises the steps:

(1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder;

(2) Take Radix Astragali, Salvia Miltiorrhiza and Ophiopogon japonicus according to the proportion, add 4 to 20 times the total weight of the three raw materials and extract with ethanol with a volume fraction of 30 to 95% for 1 to 3 times, each time for 0.5 to 3 hours, and combine The extract is filtered (or centrifuged), and the filtrate (or supernatant) is decompressed to recover ethanol, then concentrated under reduced pressure to become a thick extract with a water content of 15-20% or made into a dry-extract with a water content of 3-8% The ointment is mixed with the leech fine powder described in step (1), and a pharmaceutically acceptable carrier is added to prepare various pharmaceutical preparations.

Preparation method 3:

This preparation method comprises the steps:

Weigh Astragalus, Danshen, Ophiopogon japonicus, and leech according to the proportion, add water 4 to 20 times the total weight of the four raw materials, extract 1 to 3 times, each time for 0.5 to 3 hours, combine the extracts, filter (or centrifuge) , the filtrate (or supernatant) is concentrated under reduced pressure into a thick extract with a water content of 15-20% or made into a dry extract with a water content of 3-8%, adding pharmaceutically acceptable carriers to make various medicines preparation.

Or further, the filtrate (or supernatant) obtained by extracting Astragalus, Danshen, Ophiopogon japonicus and leech through water extraction is concentrated under reduced pressure to 1/12-1/6 of the volume of the original filtrate to obtain a concentrated solution, which is added with ethanol to contain The alcohol content is 30-90%, let it stand for precipitation, filter (or centrifuge), and after the filtrate (or supernatant) is decompressed to recover ethanol, it is concentrated under reduced pressure into a thick extract with a water content of 15-20% or made into a thick extract containing The dry extract with a water content of 3-8% is added with a pharmaceutically acceptable carrier to prepare various pharmaceutical preparations.

Preparation method 4:

This preparation method comprises the steps:

Weigh Radix Astragali, Salvia Miltiorrhiza, Ophiopogon japonicus and Leech according to the proportion, add 4 to 20 times the total weight of the four raw materials and extract with ethanol with a volume fraction of 30 to 95% for 1 to 3 times, each time for 0.5 to 3 hours, and combine the extractions liquid, filtered (or centrifuged), and after the filtrate (or supernatant) is decompressed to recover ethanol, it is concentrated under reduced pressure into a thick extract with a water content of 15-20% or a dry extract with a water content of 3-8%. , adding pharmaceutically acceptable carriers to make various pharmaceutical preparations.

Among the above four preparation methods, the extraction method of water extraction can be reflux extraction, ultrasonic extraction or immersion extraction with water at a temperature not lower than 25°C; the extraction method of ethanol extraction can be reflux extraction, ultrasonic extraction, soaking extraction or percolation extract.

In the above four preparation methods, the pharmaceutically acceptable carrier may be one or more of fillers, excipients, disintegrants and the like. The filler can be at least one of soluble starch, lactose, microcrystalline cellulose, dextrin, etc.; the excipient can be at least one of mannitol, lactose, calcium sulfate, etc.; the disintegrant can be hydroxymethyl At least one of sodium starch, modified starch, low-substituted hydroxypropyl cellulose, polyoxyethylene glycol (PEG), and the like.

Compared with the preparation method of combining and extracting the three raw materials of Astragalus, Danshen and Ophiopogon japonicus, the preparation method of combining and extracting leech into Astragalus, Salvia miltiorrhiza and Ophiopogon japonicus can significantly improve the content of each effective chemical component in the traditional Chinese medicine composition. content.

Preparation method 5:

The preparation method comprises the following steps: taking astragalus root, salvia miltiorrhiza, radices japonicus and leech according to the ratio, respectively washing, drying, pulverizing and sieving to obtain fine powder of medicinal materials, which is mixed and packed to make powder.

Using LC-MS combined technology, through DAD and MS detection, standard substance comparison, the chemical composition of the Chinese medicine composition prepared by the above preparation method is analyzed, the Chinese medicine composition of the present invention mainly contains the following chemical composition:

Flavonoids, mainly including acteosin-7-O-β-D-glucoside, formononetin, acteosin, formononetin, (6aR,11aR)-9,10-dimethoxypteroside-3 -O-β-D-glucoside and (3R)-2'-hydroxy-3',4'-dimethoxyisoflavan 7-O-β-D-glucoside, etc.;

Saponins, mainly including ophiopogon saponin, diosgenin and astragaloside IV;

Phenolic acids mainly include salvianolic acid B, salvianolic acid A, rosmarinic acid, shikonic acid, protocatechualdehyde and danshensu.

Adopt ultraviolet-visible spectrophotometry to measure, and in the Chinese medicine composition of the present invention, the weight percentage content of various chemical components is:

Taking calycosin as the reference substance, the total flavonoid content is 0.10-20.0%;

Taking salvianolic acid B as the reference substance, the total phenolic acid content is 1.0-35.0%;

Taking ruscogenin as the reference substance, the total saponin content is 0.02%-5%.

Adopt high performance liquid chromatography to measure content, by weight percentage, the content of main chemical component in the Chinese medicine composition of the present invention is respectively:

The content of calycosin-7-O-β-D-glucoside is 0.001～2%;

The content of formononetin is 0.001-1.6%;

The content of calycosin isoflavones is 0.001-2%;

The salvianolic acid B content is 0.01-10%.

By adopting the antithrombin activity method, the antithrombin activity in the traditional Chinese medicine composition of the present invention is as follows: the range of activity units is 0.1U to 10.0U.

According to another aspect of the present invention, the present invention also provides the application of the above-mentioned traditional Chinese medicine composition in the preparation of a medicament for treating myocardial infarction complicated by qi deficiency and blood stasis syndrome.

Description of drawings

Fig. 1 is a liquid chromatography-mass spectrometry (LC-MS) spectrum of the Chinese patent CN101647856B pharmaceutical composition.

Fig. 2 is the liquid chromatography-mass spectrometry of the Chinese medicine composition that the embodiment of the present invention 4 makes

(LC-MS) spectrum.

Figure 3 shows the plaque formation in the aortic sinus of ApoE-/- mice. Wherein, NOR represents the normal group; AS represents the model group; LPT represents the atorvastatin administration group; TGL, TGM, and TGH are respectively the low, middle and high dose administration groups of the traditional Chinese medicine composition of the present invention.

Fig. 4 is the comparison result of the plaque area in the aortic sinus of ApoE-/- mice. Wherein, NOR represents the normal group; AS represents the model group; LPT represents the atorvastatin administration group; TGL, TGM, and TGH are respectively the low, middle and high dose administration groups of the traditional Chinese medicine composition of the present invention.

Figure 5 shows the plaque formation in the whole section of aorta of ApoE-/- mice. Wherein, NOR represents the normal group; AS represents the model group; LPT represents the atorvastatin administration group; TGL, TGM, and TGH are respectively the low, middle and high dose administration groups of the traditional Chinese medicine composition of the present invention.

Fig. 6 is the comparison result of plaque area size in the whole section of aorta of ApoE-/- mice. Wherein, NOR represents the normal group; AS represents the model group; LPT represents the atorvastatin administration group; TGL, TGM, and TGH are respectively the low, middle and high dose administration groups of the traditional Chinese medicine composition of the present invention.

Figure 7 is a typical echocardiogram of rats in each group 5 weeks after myocardial infarction. Wherein, Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the group administered with the traditional Chinese medicine composition of the present invention.

Figure 8 is the statistical analysis results of left ventricular cardiac function parameters after 1 week (1W) and 5 weeks (5W) of myocardial infarction in rats of each group, wherein, Fig. 8B is the columnar column of statistical analysis of left ventricular ejection fraction (EF) of rats in each group Fig. 8C is a histogram of statistical analysis of fractional shortening (FS) of rats in each group, Fig. 8D is a histogram of statistical analysis of left ventricular end-diastolic inner diameter (LVIDd) of rats in each group, and Fig. 8E is a histogram of left ventricle of rats in each group Statistical analysis histogram of end-systolic inner diameters (LVIDs), . Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition administration group of the present invention; compared with the MI group, ^# P＜0.05, ^## P＜0.01; compared with the Sham group, ^* P＜0.05, ^** P<0.01.

Fig. 9 is the electrocardiogram of ventricular fibrillation induced by 8 S1 and 1 additional S2 programmed electrical stimulation in rats 5 weeks after myocardial infarction.

Fig. 10 is an electrocardiogram of non-sustained ventricular tachycardia induced by 8 S1 and 2 additional S2S3 stimulations 5 weeks after myocardial infarction in rats.

Fig. 11 is an electrocardiogram of rats that failed to induce ventricular tachycardia after 5 weeks of myocardial infarction after 8 S1 and 3 additional S2S3S4 stimulations.

Fig. 12 is the histogram of statistical analysis of ventricular arrhythmia induction rate and score after 5 weeks of rat myocardial infarction, wherein, Fig. 12D is a histogram of statistical analysis of ventricular arrhythmia induction rate of rats in each group, and Fig. 12E is a histogram of each group's large Statistical analysis histogram of rat ventricular arrhythmia score. Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition administration group of the present invention; compared with the MI group, ^# P＜0.05, ^## P＜0.01; compared with the Sham group, ^* P＜0.05, ^** P<0.01.

Fig. 13 is the representative Masson staining figure (scale bar: 100 μ m) of left ventricular myocardial fibrosis after each group of rat myocardial infarction 5 weeks, Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition of the present invention to give medicine group.

Figure 14 is a histogram of the statistical analysis of the percentage of left ventricular myocardial infarction and infarct border area fibrosis in each group of rats after 5 weeks of myocardial infarction; wherein, Figure 14B represents the histogram of the statistical analysis of the percentage of left ventricular myocardial infarction in each group of rats; Figure 14C shows the histogram of statistical analysis of the fibrosis area percentage of the left ventricular infarction border area of each group of rats; Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition administration group of the present invention; compared with the MI group, ^# P<0.05, ^## P<0.01; compared with Sham group, ^* P<0.05, ^** P<0.01.

Figure 15 shows the expression of fibroblast marker protein α-SMA in the left ventricular infarction border area of rats in each group 5 weeks after myocardial infarction. Staining representative picture (magnification is 20 times, the brown area is α-SMA staining part); Figure 15B is the statistical analysis map of α-SMA staining area in the left ventricular infarction border area, and Figure 15C is the α-SMA protein in the left ventricular infarction border area Expressing the Western blot detection results, Figure 15D is a statistical analysis chart of the α-SMA/GAPDH protein expression ratio; Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition administration group of the present invention; compared with the MI group, ^# P<0.05, ^## P<0.01; compared with Sham group, ^* P<0.05, ^** P<0.01.

Figure 16 is the serum TGF-β and MCP-1 content of rats in each group 5 weeks after myocardial infarction, wherein, Figure 16A represents the histogram of the statistical analysis of serum TGF-β content in rats of each group 5 weeks after myocardial infarction, and Figure 16B represents each Sham represents the sham operation group; MI represents the myocardial infarction model group; T represents the Chinese medicine composition administration group of the present invention; compared with the MI group, ^# P＜ 0.05, ^## P<0.01; compared with Sham group, ^* P<0.05, ^** P<0.01.

Detailed ways

The present invention will be described in further detail below by way of examples.

The following enumerates the components, proportions and preparation methods of Examples 1 to 5 of the present invention, and weighs the bulk drug (unit: parts by weight) according to the following proportioning ratio:

Example 1

Prescription: Astragalus 30 parts, Danshen 15 parts, Ophiopogon japonicus 30 parts and leeches 3 parts.

Preparation:

(1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder;

(2) Weigh astragalus, salvia miltiorrhiza and Radix Ophiopogon japonicus according to the proportion, add 10 times the total weight of the three raw materials and reflux extract 3 times, each time for 2 hours, combine the extracts, filter, and concentrate the filtrate under reduced pressure to the original filtrate 1/10 of the volume to obtain a concentrated solution;

(3) Add ethanol to the concentrated solution until the alcohol content is 60%, leave it to settle at 4°C for 24 hours, filter, and after the filtrate is decompressed to recover ethanol, it is concentrated under reduced pressure into a thick extract with a water content of 15-20%, and the steps (1) The leech fine powder is mixed evenly, added with starch to granulate, and packed into capsules to make capsules.

Example 2

Prescription: Astragalus 20 parts, Danshen 30 parts, Ophiopogon japonicus 20 parts and leech 5 parts.

Preparation:

(1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder;

(2) Take Radix Astragali, Salvia Miltiorrhiza and Ophiopogon japonicus in proportion, add 5 times of the total weight of three kinds of crude drugs, and the volume fraction is 70% ethanol reflux extraction 3 times, each 2 hours, merge extract, filter, filtrate reduces After recovering the ethanol, concentrate under reduced pressure to form a thick extract with a water content of 15-20%, mix with the leech fine powder described in step (1), add polyethylene glycol, etc. and mix evenly as the content, gelatin, glycerin, etc. Add water to dissolve as glue, and make soft capsules according to conventional methods.

Example 3

Prescription: 10 parts of Astragalus, 10 parts of Danshen, 10 parts of Ophiopogon japonicus and 3 parts of leeches.

Preparation method: weigh astragalus, salvia miltiorrhiza, Ophiopogon japonicus and leeches according to the proportion, wash, dry, pulverize and sieve respectively to obtain fine powder of medicinal materials, mix them and pack them into powders.

Example 4

Prescription: Astragalus 25 parts, Danshen 15 parts, Ophiopogon japonicus 25 parts and leeches 5 parts.

Preparation method: weigh astragalus, salvia miltiorrhiza, Ophiopogon japonicus and leech according to the proportion, add water 10 times the total weight of the four raw materials, and extract twice, 3 hours each time, combine the extracts, filter, and concentrate the filtrate under reduced pressure to obtain Thick extract with water content of 15-20%, add soluble starch, cyclodextrin, etc. to make granules.

Example 5

Prescription: 50 parts of Astragalus, 15 parts of Danshen, 15 parts of Ophiopogon japonicus and 8 parts of leeches.

Preparation method: Weigh astragalus, salvia miltiorrhiza, Ophiopogon japonicus and leeches according to the proportion, add 20 times the total weight of the four raw materials and extract with ethanol with a volume fraction of 80% for 2 times, each time for 3 hours, combine the extracts, filter, and filtrate After recovering the ethanol under reduced pressure, make a dry extract with a water content of 5%, add starch or microcrystalline cellulose, mix evenly, granulate, compress into tablets, and make tablets.

In order to prove the effect of Chinese medicine composition of the present invention, carry out following pharmacy and pharmacodynamics experiment:

Experiment 1, Chinese medicine composition of the present invention and Chinese patent CN101647856B pharmaceutical composition component liquid chromatography-mass spectrometry (LC-MS) analysis comparison

1. Drugs under test

The drug dry paste powder prepared in Example 4 of the present invention was used as the test drug sample. Take 2.09g of the test drug sample, transfer it into a 10mL measuring bottle, add ultrapure water and make it up to volume, accurately take 1mL, transfer it into a 10mL measuring bottle, and make it up to volume with ultrapure water to obtain a solution equivalent to 0.1g/mL of the crude drug.

The drug capsule prepared in Example 1 of Chinese patent CN101647856B was used as the control drug. Get 5 control drug capsules, get the contents into a 50mL measuring bottle, and adjust to volume with ultrapure water, get 1mL and transfer to a 5mL measuring bottle, and adjust to volume with ultrapure water to obtain a control drug solution.

2. High performance liquid chromatography (HPLC) and mass spectrometry conditions

Chromatographic conditions: the chromatographic column is Phenomenex Kinetex (2.1×100mm, 1.7μm). The mobile phase is composed of acetonitrile (A) and 0.1% formic acid water (B), gradient elution program: 0-2min, A (5%-17%); 2-16min, A (17%-18.4%); 16 ~22min, A (18.4%~27%); 22~45min, A (27%~37%); 45~60min, A (37%~65%). The flow rate was 400 μL·L ^-1 , the column temperature was room temperature, and the injection volume was 5 μL.

Mass spectrometry conditions: AB Triple TOF 5600 quadrupole time-of-flight mass spectrometer adopts electrospray ionization (ESI) source, spray voltage (IS): 4500V, ionization temperature (TEM): 500°C; atomization gas (GS1): 50kPa , auxiliary heating (GS2): 50kPa; curtain gas (CUR): 40kPa; collision gas (CAD): Medium; scanning mode: negative. The acquisition range of the primary mass spectrum is m/z 100-2000, the accumulation time is 250ms, the acquisition range of the secondary mass spectrum is m/z 100-1000, the accumulation time is 100ms, the collision energy (CE): 45V; the collision energy superposition (CES): 15 .

3. Experimental results:

Table 1 Chinese medicine composition of the present invention and the LC-MS analysis component content difference of reference medicine

Fig. 1 is the LC-MS spectra of the chemical components of the control drug (Fig. 1) and the Chinese medicine composition of the present invention (Fig. 2).

From the results of Fig. 1, and 2 and Table 1, it can be seen that compared with the reference drug, calycocetin-7-O-β-D-glucoside, rosmarinic acid, and formononetin in the Chinese medicine composition provided by the invention and astragaloside IV and other effective compounds significantly increased, indicating that the addition of Ophiopogon japonicus can significantly improve the content of effective chemical components of astragalus, salvia miltiorrhiza, and leech in the Chinese medicine composition of the present invention, and the effects of each component of the Chinese medicine composition of the present invention are synergistic.

Experimental example two, Chinese medicine composition of the present invention and Chinese patent CN101647856B pharmaceutical composition component content comparison

1. Tested drug: the drug tablet prepared in Example 5 of the present invention was used as the tested drug, and the drug capsule prepared in Example 1 of Chinese Patent CN101647856B was used as the control drug.

Tested drug sample Take 3 tablets of 0.3g/tablet, transfer it into a 50mL volumetric bottle, add ultrapure water and constant volume, take 1mL and transfer it into a 5mL volumetric bottle, to obtain a test drug solution. For the control drug, the contents of 3 capsules were transferred into a 50mL volumetric bottle, and the volume was fixed with ultrapure water, and 1 mL was transferred into a 5mL volumetric bottle, and the content of the ultrapure water was volumetricized to obtain the drug solution of the control capsule.

2. Experimental method

2.1 Determination method of verbascoisoflavone glycosides

Instrument: Agilent 1200 high performance liquid chromatography, chromatographic column: Agilent Zorbax SB C18, 5 μm, 4.6 × 150mm, mobile phase: acetonitrile-0.1% formic acid Gradient elution: (0～10min: 10%A; 10～20min: 10 %A→20%A), flow rate: 1mL/min, column temperature: 30°C, detection wavelength: 260nm.

2.2 Determination method of total phenolic acid

Each precision draws a certain amount of sample solution and mixed reference substance solution, each placed in a 10mL graduated test tube, each precision adds water to 2.0mL, each precision adds 0.5mL of 10% sodium nitrite solution, shakes well, and then precisely adds 10% sodium nitrite solution Aluminum nitrate solution 1.0mL, shake well, place in dark place at room temperature for 5min, add 6.0mL of 2mol/L sodium hydroxide solution, shake well, place in dark place at room temperature for 10min, use water as blank, measure absorption at 500nm wavelength Spend. Calculate the content by the standard curve method, that is, too.

3. Experimental results

Table 2 Chinese medicine composition of the present invention and contrast medicine representative component content difference

4. Experimental conclusion: as can be seen from the experimental results in Table 2, on the basis of the same clinical reference dosage, after adding Ophiopogon japonicus, the Chinese medicine composition of the present invention is compared with the reference drug, and the content of representative component acteosin glucoside is obvious. increased, while the total phenolic acid content decreased significantly. This shows that the Chinese medicine composition of the present invention and the Chinese patent CN101647856B pharmaceutical composition show obvious differences in representative components. big difference.

Experimental Example 3: Experimental study on the toxicity of the traditional Chinese medicine composition of the present invention to ICR mice after single oral administration 1. Dosage design: This experiment intends to use the maximum dosage method. The medicine dry ointment powder of the embodiment of the present invention 4 is used as the test drug, and the dose converted into mice according to the body surface area of the reference clinical dose is: 2.16g/kg of raw medicinal materials, 0.60g/mL is the test drug dry ointment powder The maximum drug concentration that can pass through the mouse gavage needle smoothly. Through pre-experiment, it is determined that 48g/kg dry paste powder of the test drug is the maximum dosage, and both the negative reference substance and the vehicle are 0.5% carboxymethylcellulose sodium (CMC-Na).

2. Experimental grouping and administration: 40 SPF-grade ICR mice qualified for quarantine were selected and randomly divided into two groups, namely the negative control group and the test drug dry cream powder group, with 20 mice in each group, half male and half male. At least 12 hours of fasting before administration, no water, 40g/kg intragastric administration, once in the morning and once in the afternoon (interval greater than 4h). Observe for 4 hours after administration, and then observe at least once a day for 14 days. The body weight of the animals was measured on the 2nd, 3rd, 5th, 7th, 11th, and 14th days after administration. Dead animals were also weighed.

3. Detection indicators: general observations include changes in animal weight, diet, appearance, behavior, secretions, excretions, etc. Pathological examinations included gross dissection of poisoned dead or dying animals in time, and dislocation of other animals after the end of the observation period for gross dissection. Gross anatomical examination organs include trachea, esophagus, heart, liver, spleen, lung, kidney, adrenal gland, thymus, brain, pancreas, stomach, intestine (duodenum, jejunum, ileum, cecum, colon, rectum), mesenteric lymph nodes , bladder, prostate, testis (with epididymis), uterus, ovary.

4. Experimental results: No animal death was found in the experimental mice in each group, and no pathological abnormal changes were found in the naked eyes and pathological sections of the organs examined in the mice. 14/20 animals (10♂, 4♀) had loose stools after intragastric administration of 48g/kg dry cream powder within 30min to 4h after administration, and 3 animals (2♂, 1♀) ) recovered within 1 hour after drug administration, 1 animal (♀) recovered within 2 hours after drug treatment, and the remaining 10 animals did not recover within 4 hours after drug treatment, and recovered on the next day after drug administration. Compared with the negative control group, the test drug dry paste powder 48g/kg administered the next day can cause the male animals to slow down the body weight growth, and the difference is statistically significant (P<0.05). However, there was no statistical difference in the body weight at other time points compared with the negative control group, and the body weight maintained a normal growth trend within 14 days of the observation period. The body weight of the female animals maintained a normal growth trend within 14 days of the observation period.

5. Experimental conclusion: When the dry ointment powder of the test drug was 48g/kg (converted to 1001 times the clinically intended dosage in terms of kilogram body weight) as the maximum dosage, no obvious toxic reaction in mice occurred. The slow weight gain of male animals on the second day of administration may be related to the loose stools of animals on the day of administration, which affects the weight gain of animals. The traditional Chinese medicine composition of the invention has low toxicity and good safety.

Experimental example 4, experimental research on the effect of Chinese medicine composition of the present invention on ApoE ^-/- mice atherosclerosis

1. Experimental drug: adopt the granules prepared in Example 4 of the present invention, and the high, medium and low dosages are converted into mouse equivalent dosages of 0.8 times, 1.5 times and 2.5 times according to the clinical dosage. The positive control was atorvastatin, which was dissolved with 0.5% sodium carboxymethylcellulose (CMC-Na) to the desired concentration.

2. Experimental animals and grouping: ApoE ^-/- mice (strain C57BL/6J, SPF grade, 5 weeks old), male, weighing 25±5 g, were grouped according to the following experiment, 15 mice in each group.

Normal group: ApoE ^-/- mice were given normal feed for 12 weeks;

Model group: ApoE ^-/- mice were fed with high-fat diet and fed with 0.2mL/10g normal saline every day for 12 weeks;

Atorvastatin administration group: ApoE ^-/- mice were fed with high-fat diet, and at the same time were given an equal volume (0.2mL/10g) of atorvastatin aqueous solution (10mg/kg) orally every day for 12 weeks;

Experimental drug administration group: ApoE ^-/- mice were fed with high-fat feed, and the high, middle, and low dose administration groups were fed with an equal volume (0.2mL/10g) of the corresponding aqueous solution of the traditional Chinese medicine composition at the same time for 12 days. week.

3. Experimental index detection

3.1 Determination of blood lipids: After the experiment, blood was collected from the orbit of the mice to separate serum, and the contents of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were detected;

3.2 Oil red O staining of the entire aorta and the blood vessels at the aortic valve: after blood collection from the mice, the entire aorta (from the heart to the bifurcation of the common iliac artery) and the blood vessels at the aortic valve were separated and washed with PBS. After removing the remaining fat and connective tissue, fix it in 4% paraformaldehyde for more than 12 hours, then cut it longitudinally, with the inner cavity facing upward, soak in 70% ethanol for 2 minutes, stain with Oil Red O for 30 minutes, rinse with 70% ethanol until the tissue Turn white, take pictures after several times of distilled water, and use software to quantitatively analyze the plaque area.

3.3 Detection of mouse serum adiponectin (Adiponectin), monocyte chemoattractant-1 (MCP-1), tumor necrosis factor-α (TNF-α) content: After the experiment, blood was collected from the mouse orbit to separate serum, and The contents of the above indicators were detected by Elisa method.

4. Experimental results:

4.1 Determination of blood lipids: The data in Table 3 show that the serum TC content of the model group mice is significantly higher than that of the normal group. After treatment with atorvastatin, high dose and medium dose of the test drug, the serum TG content of the mice decreased significantly; the serum LDL-C content of the mice in the model group was significantly higher than that of the normal group, while the high dose and medium dose Dosage test drug group can significantly reduce serum LDL-C content. There was no significant difference in serum HDL-C levels among the groups.

Table 3 Detection of blood lipid levels in mice of each group

NOR group: normal group; AS group: model group; LPT group: atorvastatin administration group. ^* or ^** P<0.05 or P<0.01vs NOR group; ^{#or ##} P<0.05 or P<0.01vs AS model group.

4.2 Mouse aorta pathology: see Figure 3-6, after 12 weeks of high-fat feeding of ApoE-/- mice, oil red O staining of the aortic sinus and aorta showed obvious atherosclerosis in the model group After 8 weeks of administration, the test drug can significantly inhibit the formation of aortic sinus plaque induced by high-fat diet, and the area of plaque in the aortic sinus and the entire aorta is significantly reduced. The number of plaques was large, and there was no statistical difference between the plaque area and the model group.

4.3 Detection of mouse serum adiponectin (Adiponectin), MCP-1, TNF-α content: adiponectin is a hormone protein secreted specifically by mature adipocytes, which can enhance insulin sensitivity, regulate glucose and lipid metabolism, resist Inflammatory and anti-atherosclerotic effects. In the serum of patients with atherosclerosis, the content of adiponectin decreased instead. MCP-1 can be secreted by a variety of cells, and it plays an important role in allergy, immunodeficiency disease, kidney disease, atherosclerosis and other diseases. MCP-1 is elevated in macrophage-rich atherosclerotic plaques in humans and primates and recruits monocytes to migrate into atherosclerotic plaques.

The results in Table 4 show that the traditional Chinese medicine composition of the present invention can significantly inhibit the decrease of serum adiponectin content and the increase of serum MCP-1 content in mice caused by high-fat diet, and the effect is better than the positive control drug atorvastatin.

Table 4 Mouse serum adiponectin, MCP-1, TNF-α content determination results

NOR group: normal group; AS group: model group; LPT: group atorvastatin administration group. ^* or ^** P<0.05 or P<0.01vs NOR group; ^{#or ##} P<0.05 or P<0.01vs AS model group.

5. Experimental conclusion: The traditional Chinese medicine composition of the present invention can significantly reduce the levels of serum TG, TC and LDL-C in atherosclerotic mice, and reduce the levels of inflammatory factors related to atherosclerosis. The medium and high doses of the composition can significantly inhibit the induction of high-fat diet. aortic sinus plaque formation, indicating that the traditional Chinese medicine composition of the present invention has obvious anti-atherosclerotic effect in mice.

Experimental Example 5, Experimental Research on the Myocardial Remodeling of the Chinese Medicine Composition of the Present Invention on Rats After Myocardial Infarction . 8-week-old male SD rats were selected. After anesthesia, a non-invasive endotracheal intubation was used to connect to a ventilator. The heart was exposed by opening the chest, and the ligature was passed through the LAD (2-3 mm below the left atrium). Successful myocardial ischemia was confirmed by visual observation (pale color) and ECG monitoring (ST-segment elevation and QRS widening). The sham operation group received the same operation process, but the thread under the LAD was not ligated.

2. Administration and grouping of rats: rats were divided into three groups, sham operation group (sham), myocardial infarction model group (MI), and experimental drug group (T). Referring to the literature of Lee (Circulation, 2006), in order to ensure the uniformity of myocardial infarction size, the model group and the administration group selected rats with an echocardiographic EF<45% after 1 week, and then randomly divided them into groups. The model group was given 0.5% carboxymethyl fiber Sodium plain (CMC-Na) aqueous solution gavage, administration group adopts the Chinese medicine composition extract that the embodiment of the present invention 2 makes, dissolves to 620mg/kg/ with 0.5% carboxymethylcellulose sodium (CMC-Na) d Oral administration for 28 consecutive days.

3. Detection indicators

3.1 Ultrasound examination of the rat heart: Ultra-high-resolution small animal ultrasound imaging system (Vevo2100) was used. After anesthesia, a high-frequency ultrasound probe (21MHz) was used to continuously collect dynamic images of 10 cardiac cycles in 4 different sections. Each group of data The average value of 5 consecutive cardiac cycles was taken to measure the left ventricular end-diastolic inner diameter (LVIDd), left ventricular end-systolic inner diameter (LVIDs), left ventricular pressure (LVP), cardiac function indexes (left ventricular ejection fraction (LVEF)%, Left ventricular fractional shortening (LVFS) %, cardiac output (SV) value) are calculated by the built-in software.

3.2 Detection of susceptibility to ventricular arrhythmia: using programmed electrical stimulation to induce ventricular arrhythmia and synchronously recording the electrocardiogram: hook the positive pole of the electrode to the peripheral area of the infarction, and place the electrode in the sham operation group on the corresponding part of the heart. The pulse width of stimulation was 2ms, and the pacing threshold was measured. If ventricular arrhythmia occurred during the test threshold, the experiment was terminated, and the scores were scored according to the standard. The intensity of electrical stimulation was twice the pacing threshold, and the S1S2S3S4 stimulation method was used. After 8 basic stimulations, a pre-procedural stimulation was performed, starting with S1S2=120ms, and the step length was -4ms, until the induction or refractory period was reached. If the refractory period is reached, S3 and S3S4 are sequentially added before the period. Referring to the literatures of Nguyen (Circulation, 1998) and Xin (Eur J Pharmacol, 2010), the specific scoring criteria are shown in the table below. The higher the score, the higher the susceptibility to ventricular arrhythmia. The ERP stimulated by S1S1=120ms and S1S2 was used as the ERP of the ventricles of rats.

Table 5 Scoring criteria for ventricular arrhythmia

3.3 Determination of transforming growth factor-β (TGF-β) and monocyte chemoattractant-1 (MCP-1) in rat serum: 5 weeks after myocardial infarction, the abdominal aortic blood was anesthetized and centrifuged to prepare serum. Elisa kit manual to measure serum TGF-β and MCP-1 content;

3.4 Rat ventricular Masson staining and α-smooth muscle actin (α-SMA) histochemical detection: Rat left ventricular samples were fixed in 10% formaldehyde solution, embedded in paraffin, sectioned, and stained with Masson to detect the left side of the myocardial infarction area. The change of ventricular fibrosis, the degree of myocardial interstitial fibrosis was reflected by the integral of collagen volume. The area of fibroblast marker protein α-SMA detected by immunohistochemistry represents the content of myocardial fibroblasts.

3.5 Detection of α-SMA protein expression in the left ventricular infarction border area of rats: The myocardial tissue in the left ventricular infarction border area of rats in each group was collected, and Western blot method was used to detect the expression of α-SMA protein.

4. Experimental results

4.1 The effect of the traditional Chinese medicine composition of the present invention on the improvement of left ventricular cardiac function after 5 weeks of myocardial infarction in rats: as shown in Figure 7 and Figure 8, after 1 week of myocardial infarction, the cardiac function of the MI group and the administration group before administration Parameters—left ventricular ejection fraction (EF), fractional shortening (FS), left ventricular end-systolic and end-diastolic diameters (LVIDs, LVIDd) were significantly lower than those in the sham group, and there were no significant differences between the two groups. After 5 weeks, the left ventricular ejection fraction (EF) and short-axis shortening (FS) of the administration group were significantly higher than the MI group, and the left ventricular diameter (LVID) was significantly lower than the MI group, indicating that the Chinese medicine composition of the present invention can obviously improve myocardial Effect of left ventricular function 5 weeks after infarction.

4.2 The effect of the traditional Chinese medicine composition of the present invention on the score of ventricular arrhythmia after 5 weeks of myocardial infarction in rats: the results are shown in Figures 9-12. After 5 weeks of myocardial infarction, 86.7% of the rats (13/15) in the model group induced persistent ventricular premature beats, ventricular tachycardia or ventricular fibrillation, and the average score of ventricular arrhythmia was 5, while only 46.7% of the rats in the treatment group Rats (7/15) had arrhythmia, and most of them needed to give S1S2S3 stimulation to induce premature ventricle, and the average value of arrhythmia score was reduced to 3.6, which shows that the Chinese medicine composition of the present invention can obviously reduce the ventricular rhythm of rat myocardial infarction in chronic phase. Disorder susceptibility.

4.3 The effect of the traditional Chinese medicine composition of the present invention on myocardial fibrosis after 5 weeks of myocardial infarction in rats: the results are shown in Fig. 13 and Fig. 14 . After 5 weeks of myocardial infarction, the myocardial infarction area in the model group accounted for 42% ± 3.6%, and the area of fibrosis in the peri-infarct area was 46.3 ± 3.5%. %, significantly decreased compared with the model group.

4.4 The effect of the traditional Chinese medicine composition of the present invention on myocardial fibroblast marker protein α-SMA in the peri-infarct area of rat myocardial infarction 5 weeks later: the results are shown in FIG. 15 . The average area of α-SMA positive cells in the infarct peripheral area of the model group was 18.6%, and the average area of α-SMA positive cells in the treatment group was 10.4%, which was significantly lower than that of the model group, and the expression of α-SMA protein was also significantly reduced, both statistically significant. difference.

4.5 The effect of the traditional Chinese medicine composition of the present invention on the levels of serum TGF-β and MCP-1 in rats 5 weeks after myocardial infarction: the results are shown in FIG. 16 . After 5 weeks of myocardial infarction, the levels of TGF-β and MCP-1 in the serum of the model group were 1277.6pg/mL and 2398.9pg/mL, while the levels of TGF-β and MCP-1 in the serum of the treatment group were lower than those of the model group. It is statistically significant.

5. Experimental conclusion: the traditional Chinese medicine composition of the present invention can significantly improve the left ventricular cardiac function after 5 weeks of myocardial infarction in rats, reduce the induction rate and arrhythmia score of ventricular arrhythmia after myocardial infarction, and improve the ventricular infarction area and infarction periphery. The degree of tissue fibrosis in the region, its mechanism is related to the reduction of inflammatory factors by the Chinese medicine composition of the present invention, and the inhibition of myocardial myofibroblast transformation and myocardial fibrosis. This shows that the Chinese medicine composition of the present invention inhibits myocardial remodeling (anti-myocardial remodeling) after myocardial infarction. Myocardial fibrosis), promoting the recovery of heart function, and reducing the induction of severe ventricular arrhythmia have obvious effects.

In the experiment of myocardial remodeling after myocardial infarction in rats, the effective dose of the Chinese medicine composition of the present invention is 620 mg/kg, which is far lower than the effective dose of 10 g/kg of the pharmaceutical composition disclosed in CN101647856B using the same test method, indicating that the Chinese medicine composition of the present invention The composition has remarkable medicinal effect, low therapeutic dosage and is suitable for long-term administration.

Experimental example 6. Clinical study of Chinese medicine composition of the present invention on patients with myocardial infarction with Qi deficiency and blood stasis syndrome combined with cardiac insufficiency (Qi deficiency and blood stasis type heart failure)

1. Case source: In order to further demonstrate the clinical effect of the traditional Chinese medicine composition of the present invention on the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome, a prospective randomized controlled study was carried out according to the requirements of evidence-based medicine. 60 patients with cardiac dysfunction after myocardial infarction were used as clinical research objects, including 30 cases in each of the Chinese medicine composition treatment group and the control group of the present invention, and the follow-up time: 0 to 3 months.

2. Inclusion and exclusion criteria

Inclusion criteria: ①Aged between 18 and 80 years old with cardiac insufficiency, in line with Killip grade II-III after acute myocardial infarction or reference plasma BNP level; ②with clinical manifestations of qi deficiency and blood stasis, dialectically as qi deficiency and blood stasis syndrome ;

Exclusion criteria: ① Those with mechanical complications and cardiogenic shock before inclusion; ② Chronic heart failure caused by heart valve disease, congenital heart disease, pericardial disease, arrhythmia or other non-cardiac causes; ③ Severe Patients with liver, kidney, nerve, and blood coagulation disorders, patients with COPD and gout, and those who cannot complete the 6-minute walk test due to waist and leg diseases;

3. Intervention plan

3.1 Western medicine treatment:

(1) Nitrate drugs: nitroglycerin, isosorbide mononitrate (Xinkang);

(2) ACEI/ARB: captopril, candesartan, valsartan, fosinopril, irbesartan;

(3) Receptor blockers: metoprolol tartrate, betaloc, bisoprolol, metoprolol succinate;

(4) Cardiotonic agents: deacetyllanatoside, digoxin.

During the treatment period, the selected patients are not allowed to take traditional Chinese medicine decoctions and other Chinese patent medicines. For patients with primary diseases, symptomatic treatment can be carried out according to current guidelines.

3.2 Intervention treatment

On the basic treatment of western medicine standard, the patients in the treatment group started to take the granules prepared by the embodiment of the present invention 4 orally on the same day; The placebo granules whose texture etc. are consistent with the granules of the present invention. Dosage: 1 bag (6 grams) per time, 3 times a day with boiled water before meals, for 3 consecutive months.

4. Efficacy evaluation: TCM symptom evaluation, 6-minute walking distance, heart enzyme, troponin value, BNP content, NYHA cardiac function classification, qi deficiency and blood stasis syndrome score, safety observation and evaluation.

5. Experimental results

In this study, a total of 60 patients with myocardial infarction and PCI combined with cardiac insufficiency participated in this study, including 30 patients in the treatment group (accounting for 50%), and 30 patients in the control group (accounting for 50%), of which 52 patients Complete this study, including 25 cases in the control group and 27 cases in the treatment group.

5.1 Baseline data

A total of 60 patients were included in this study. There were 24 males and 6 females in the treatment group; 26 males and 4 females in the control group.

Table 6 Demographic data of patients in the two groups

5.2 Comparison of cardiac function classification and curative effect between the two groups

After 3 months of treatment, the heart function of the treatment group was significantly improved compared with the control group, and the difference was statistically significant (P=0.009; P<0.05). The results are shown in Table 7 and Table 8.

Table 7 Comparison of NAYH cardiac function classification between the two groups

5.3 Comparison of Qi Deficiency and Blood Stasis Syndrome Scale Scores between the two groups

There was no significant difference in the score of Qi deficiency and blood stasis syndrome scale between the treatment group and the control group before treatment (P=0.957; P>0.05); The effective rate was 81.2%, and that of the control group was 72%, with significant difference (P=0.007; P<0.05). The results are shown in Table 9 and Table 10.

Table 9 Comparison of the effective rate of Qi Deficiency and Blood Stasis Syndrome Scale scores between the two groups

Table 10 Comparison of Qi Deficiency and Blood Stasis Syndrome Scale scores between the two groups

5.4 Analysis of the 6-minute walking distance of the two groups

The 6-minute walking distance of the two groups is shown in Table 11 and Table 12. Although the walking distance of the treatment group was not significantly different from that of the control group after treatment, there was a significant difference in the walking distance of the two groups after treatment (P=0.009 ; P<0.05).

Table 11 Comparison of 6-minute walking distance of two groups of patients

Table 12 Comparison of 6-minute walking distance increment before and after treatment in two groups of patients

5.5 Comparison of readmission rates

There was a statistically significant difference in the rehospitalization rate and mortality rate between the two groups within 30 days of admission (P<0.05). The results are shown in Table 13.

Table 13 Comparison of the readmission rate of two groups of patients within 3 months (cases)

5.6 Comparison of liver and kidney function between the two groups before and after treatment

There was no significant change in the liver and kidney function indexes of the two groups of patients before and after treatment (P>0.05). The results are shown in Table 14 and Table 15.

Table 14 Comparison of liver function before and after treatment in two groups of patients

Table 15 Comparison of renal function between the two groups of patients before and after treatment

6. Experimental conclusion: the traditional Chinese medicine composition provided by the present invention can improve the subjective heart function-related symptoms of patients with myocardial infarction complicated by qi deficiency and blood stasis syndrome, and improve the symptoms of qi deficiency and blood stasis in patients with acute myocardial infarction after PCI without obvious It has a significant curative effect on the liver and kidney damage function of Qi deficiency and blood stasis type myocardial infarction combined with heart failure.

What have been described above are only some embodiments of the present invention. For those skilled in the art, without departing from the inventive concept of the present invention, several modifications and improvements can be made, and these all belong to the protection scope of the present invention.

Claims (10)

1. The traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome, characterized in that its active ingredients are made of the following raw materials in parts by weight: 5-60 parts of astragalus, 5-60 parts of salvia miltiorrhiza, 5 parts of Ophiopogon japonicus ～60 parts and 3～10 parts of leeches. 2. The traditional Chinese medicine composition for the secondary prevention of myocardial infarction due to qi deficiency and blood stasis according to claim 1, wherein the active ingredients are made of the following raw materials by weight: 10-50 parts of Radix Astragali, 10 parts of Salvia Miltiorrhiza ～30 parts, 10～50 parts of Ophiopogon japonicus and 3～10 parts of leeches. 3. The traditional Chinese medicine composition for the secondary prevention of myocardial infarction due to qi deficiency and blood stasis according to claim 2, wherein its active ingredients are made of the following crude drugs in parts by weight: 25 parts of astragalus, 15 parts of salvia miltiorrhiza, 25 parts of Ophiopogon japonicus and 5 parts of leeches. 4. The traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome according to any one of claims 1 to 3, characterized in that, the traditional Chinese medicine composition is tablet, capsule, powder, oral liquid, Pills, ointments, elixirs, granules or granules. 5. The preparation method of the traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome according to any one of claims 1 to 3, characterized in that the preparation method comprises the following steps: (1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder; (2) Weigh astragalus, salvia miltiorrhiza and Ophiopogon japonicus according to the proportion, add 4 to 20 times the total weight of the three raw materials to extract 1 to 3 times, each time for 0.5 to 3 hours, combine the extracts, filter, and the filtrate reduces Concentrate under pressure to 1/12 to 1/6 of the volume of the original filtrate to obtain a concentrated solution; (3) Add ethanol to the concentrated solution until the alcohol content is 30-90%, let it stand for precipitation, filter, and after the filtrate is decompressed to recover ethanol, it is concentrated under reduced pressure into a thick extract with a water content of 15-20% or made into a water content 3-8% dry extract, mixed with the leech fine powder described in the step (1), adding pharmaceutically acceptable carriers to prepare a pharmaceutical preparation. 6. The preparation method of the traditional Chinese medicine composition for the secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome according to any one of claims 1 to 3, characterized in that the preparation method comprises the following steps: (1) Take the leech according to the proportioning ratio, wash, dry, pulverize, and sieve to obtain the leech fine powder; (2) Take Radix Astragali, Salvia Miltiorrhiza and Ophiopogon japonicus according to the proportion, add 4 to 20 times the total weight of the three raw materials and extract with ethanol with a volume fraction of 30 to 95% for 1 to 3 times, each time for 0.5 to 3 hours, and combine The extract is filtered, and after the ethanol is reclaimed from the filtrate under reduced pressure, it is concentrated under reduced pressure into a thick extract with a water content of 15-20% or a dry extract with a water content of 3-8%. The fine powder is mixed evenly, and a pharmaceutically acceptable carrier is added to make a pharmaceutical preparation. 7. The preparation method of the traditional Chinese medicine composition for the secondary prevention of myocardial infarction according to any one of claims 1 to 3, characterized in that the preparation method comprises the following steps: weighing Astragalus membranaceus, Salvia miltiorrhiza, Radix Ophiopogon japonicus and leech, add water of 4 to 20 times the total weight of the four raw materials and extract 1 to 3 times, each time for 0.5 to 3 hours, combine the extracts, filter, and concentrate the filtrate under reduced pressure to a water content of 15 ~20% thick extract or made into dry extract with a water content of 3-8%, adding pharmaceutically acceptable carriers to make a pharmaceutical preparation. 8. The preparation method of the traditional Chinese medicine composition for the secondary prevention of myocardial infarction due to qi deficiency and blood stasis according to any one of claims 1 to 3, characterized in that the preparation method comprises the following steps: weighing astragalus, Salvia miltiorrhiza, Ophiopogon japonicus and leech, add 4 to 20 times the total weight of the four raw materials and extract with ethanol with a volume fraction of 30 to 95% for 1 to 3 times, each time for 0.5 to 3 hours, combine the extracts, filter, and reduce the filtrate After recovering the ethanol, concentrate under reduced pressure to form a thick extract with a water content of 15-20% or make a dry extract with a water content of 3-8%, and add pharmaceutically acceptable carriers to make various pharmaceutical preparations. 9. The preparation method of the traditional Chinese medicine composition for secondary prevention of myocardial infarction due to qi deficiency and blood stasis syndrome according to any one of claims 1 to 3, characterized in that the preparation method comprises the following steps: weighing Astragalus membranaceus, Salvia miltiorrhiza, Radix Ophiopogon japonicus and leeches are respectively washed, dried, pulverized, and sieved to obtain fine powder of medicinal materials, which are mixed evenly and subpackaged to make a powder. 10. The use of the traditional Chinese medicine composition for secondary prevention of myocardial infarction with qi deficiency and blood stasis syndrome according to any one of claims 1 to 3 in the preparation of a drug for treating myocardial infarction with qi deficiency and blood stasis syndrome combined with cardiac insufficiency.