CN109568448A - A kind of Chinese medicine composition and preparation method thereof for treating acute coronary syndrome - Google Patents

A kind of Chinese medicine composition and preparation method thereof for treating acute coronary syndrome Download PDF

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CN109568448A
CN109568448A CN201910007680.2A CN201910007680A CN109568448A CN 109568448 A CN109568448 A CN 109568448A CN 201910007680 A CN201910007680 A CN 201910007680A CN 109568448 A CN109568448 A CN 109568448A
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parts
chinese medicine
radix
medicine composition
water
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邓悦
于秀华
陈声武
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AFFILIATED HOSPITAL TO CHANGCHUN UNIVERSITY OF CHINESE MEDICINE
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Abstract

The present invention provides a kind of Chinese medicine compositions and preparation method thereof for treating acute coronary syndrome, belong to acute coronary syndrome treatment technology field, the Chinese medicine composition includes the raw material of following parts by weight: 20~50 parts of honeysuckle, 20~50 parts of root of kirilow rhodiola, 5~20 parts of leech, 10~50 parts of Snakegourd Fruit, 10~30 parts of radix scrophulariae, 20~40 parts of Radix Angelicae Sinensis, 10~30 parts of rhizoma nardostachyos, 20~40 parts of Radix Salviae Miltiorrhizae, 5~10 parts of Radix Glycyrrhizae.The Chinese medicine composition is mainly used for the treatment of coronary heart disease after coronary heart disease unstable angina, Non-ST Elevation Acute type myocardial infarction, hematological reconstitution, and the adjuvant treatment of ST sections of elevation myocardial infarctions, preclinical pharmacodynamics of San research confirms that the Chinese medicine composition can improve the myocardial ischemia of Experimental Rats of Myocardial Ischemia model, with anti-oxidative stress, the index of myocardium enzyme and myocardial injury markers can be improved, function of vascular endothelium can be improved, have the effects that anti-inflammatory, stable patch.

Description

A kind of Chinese medicine composition and preparation method thereof for treating acute coronary syndrome
Technical field
The present invention relates to acute coronary syndrome treatment technology field more particularly to a kind of acute coronary syndromes for the treatment of Chinese medicine composition and preparation method thereof.
Background technique
Acute coronary syndrome (ACS) be with Coronary Atherosclerotic Plaque rupture or invade, it is secondary completely or Incomplete occluding thrombus is formed as one group of clinical syndrome of pathologic basis, including Acute pathologic and chronic health evaluation, urgency Property non-ST segment elevation myocardial infarction and unstable angina pectoris (UA).
ACS is a kind of common serious cardiovascular disease, is a kind of serious types of coronary heart disease.It is common in old, male Property and postmenopausal women, smoking, hypertension, diabetes, hyperlipidemia, Central obesity and the trouble for having premature coronary heart disease family history Person.ACS patient often appears as the symptoms such as ictal pectoralgia, uncomfortable in chest, can lead to arrhythmia cordis, heart failure, even dies suddenly, sternly Ghost image rings quality of life and the service life of patient.If taken appropriate therapeutic modality in time, then case fatality rate can be substantially reduced, and reduce Complication improves the prognosis of patient.
It at this stage, mainly include thromboembolism treatment, percutaneous coronary intervention for the treatment of acute coronary syndrome (PCI) it treats and carries out antithrombotic treatments and the treatment etc. that resists myocardial ischemia by taking western medicine class drug.But still some Crowd, it is necessary to have the tcm products of clinical effectiveness the reason of because of intervention time window and economic condition.And the Chinese medicine being currently known Product is largely confined to the treatment of this hypotype of coronary disease and angina pectoris, without for including that the coronary heart disease unstability heart twists Bitterly, in the complete treatment of the acute coronary syndrome after Non-ST Elevation Acute type myocardial infarction and hematological reconstitution including coronary heart disease Medicine product.
Summary of the invention
The purpose of the present invention is to provide a kind of Chinese medicine compositions and preparation method thereof for treating acute coronary syndrome, should Chinese medicine composition can be used in coronary disease after coronary heart disease unstable angina, Non-ST Elevation Acute type myocardial infarction and hematological reconstitution The treatment of disease and the adjuvant treatment of ST sections of elevation myocardial infarctions.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of Chinese medicine composition for treating acute coronary syndrome, the raw material including following parts by weight: 20~50 parts of honeysuckle, 20~50 parts of root of kirilow rhodiola, 5~20 parts of leech, 10~50 parts of Snakegourd Fruit, 10~30 parts of radix scrophulariae, Radix Angelicae Sinensis 20~ 40 parts, 10~30 parts of rhizoma nardostachyos, 20~40 parts of Radix Salviae Miltiorrhizae and 5~10 parts of Radix Glycyrrhizae.
Preferably, the Chinese medicine composition includes the raw material of following parts by weight: 30~40 parts of honeysuckle, root of kirilow rhodiola 30~40 Part, 10~15 parts of leech, 20~40 parts of Snakegourd Fruit, 15~25 parts of radix scrophulariae, 25~35 parts of Radix Angelicae Sinensis, 15~25 parts of rhizoma nardostachyos, Radix Salviae Miltiorrhizae 25~ 35 parts and 6~9 parts of Radix Glycyrrhizae.
The present invention provides the preparation methods of the Chinese medicine composition described in above scheme, comprising the following steps:
1) Radix Salviae Miltiorrhizae being mixed with ethanol water, carries out refluxing extraction, filtering collects filtrate concentration, obtains the first thick paste, Collection filter residue is first dregs of a decoction；
2) honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos are mixed with water, carry out the first water extraction, collection obtain volatile oil, extracting solution and Second dregs of a decoction；
3) by step 1) first dregs of a decoction, step 2) second dregs of a decoction and root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae, Radix Glycyrrhizae It is mixed with water, carries out the second water extraction, filtering, concentrated filtrate obtain molten cream；
4) it stands, extracts after mixing step 3) the molten cream with ethanol water, filtering, concentrated filtrate obtain Second thick paste；
5) quiet after mixing step 1) first thick paste, step 4) second thick paste and the step 2) extracting solution It sets, filtering, dried filtrate, dried object crush to obtain medicinal powder；
6) by volatile oil beta-cyclodextrin inclusion compound described in step 2), inclusion compound is obtained；
7) inclusion compound described in step 6) is mixed with medicinal powder described in step 5), obtains Chinese medicine composition.
There is no temporal restriction relations for step 5) and step 6).
Preferably, the mass ratio of Radix Salviae Miltiorrhizae and ethanol water is 1:(7~9 in the step 1)).
Preferably, the percent by volume of ethyl alcohol is 80%~95% in ethanol water in the step 1).
Preferably, the temperature of the refluxing extraction in the step 1) is 60~80 DEG C；The number of the refluxing extraction be 2~ 4 times.
Preferably, the mass ratio of honeysuckle, the gross mass of Radix Angelicae Sinensis and rhizoma nardostachyos and water is 1:(9~11 in the step 2)).
Preferably, the temperature that the second water extracts in the step 3) is 80~100 DEG C；The number that second water extracts is 2~4 times；The time extracted every time is 1~3h.
Preferably, the temperature stood in the step 5) is 1~5 DEG C；The time of the standing is 18~30h.Preferably, The partial size of the step 5) traditional Chinese medicine powder is 80~100 mesh.
Beneficial effects of the present invention: the present invention provides a kind of Chinese medicine composition for treating acute coronary syndrome and its systems Preparation Method, the Chinese medicine composition include the raw material of following parts by weight: 20~50 parts of honeysuckle, 20~50 parts of root of kirilow rhodiola, leech 5~ 20 parts, 10~50 parts of Snakegourd Fruit, 10~30 parts of radix scrophulariae, 20~40 parts of Radix Angelicae Sinensis, 10~30 parts of rhizoma nardostachyos, 20~40 parts of Radix Salviae Miltiorrhizae and Radix Glycyrrhizae 5 ~10 parts；The Chinese medicine composition is removed obstruction in channels to relieve pain with promoting blood circulation of detoxifying as therapeutic rules.Wherein, honeysuckle, property is sweet cold, is good at heat-clearing Removing toxic substances, therefore reusing is main ingredient；Its is sweet in flavor and weigh for Radix Angelicae Sinensis, is apt to promoting blood circulation to remove blood stasis；Its sweet in flavor, bitter cold of radix scrophulariae, the function with purging fire for removing toxin； Radix Salviae Miltiorrhizae, bitter slightly cold, promoting blood circulation to remove blood stasis；Radix Glycyrrhizae gas sweet taste has the benefits of removing summer-heat hundred is malicious, reinforce with honeysuckle it is clearing heat and detoxicating it Power is to assist altogether.Complete square compatibility 5, clearly without dry, cold and not clot, profit it is oiliness, grow and can lead to, play to detoxify altogether and invigorate blood circulation, dredging collateral stops The function of pain.Add keeps its full side's removing toxic substances promoting blood circulation effect stronger with drugs such as Radix Salviae Miltiorrhizaes.Chinese medicine composition of the invention is mainly used for coronary disease The treatment of coronary heart disease after sick unstable angina, Non-ST Elevation Acute type myocardial infarction, hematological reconstitution, preclinical pharmacodynamics of San research It confirms that the Chinese medicine composition can improve the myocardial ischemia of Experimental Rats of Myocardial Ischemia model, there is anti-oxidation stress to make With the index of myocardium enzyme and myocardial injury markers can be improved, by showing the Chinese medicine composition to its study on mechanism Function of vascular endothelium can be improved, have the effects that anti-inflammatory, stable patch.
Detailed description of the invention:
Fig. 1 shows the preparation method flow charts of Chinese herbal granules；
Fig. 2 indicates the case where one center creatase of experiment of embodiment 4 and Applications of Cardiac Markers are expressed；
Fig. 3 indicates the situation of change in the experiment two of embodiment 4 before modeling with electrocardiogram after modeling；
Fig. 4 indicates the myocardial infarct size of different experiments group in the experiment two of embodiment 4；
Fig. 5 indicates the case where two center creatase of experiment of embodiment 4 and Applications of Cardiac Markers are expressed；
Fig. 6 indicates the myocardial pathological morphology of different experiments group in the experiment two of embodiment 4, and wherein Fig. 6-a is sham-operation group Myocardial pathological morphology；Fig. 6-b is model group myocardial pathological morphology；Fig. 6-c is low dose group myocardial pathological morphology；During Fig. 6-d is Dosage group myocardial pathological morphology；Fig. 6-e is high dose group myocardial pathological morphology；6-f is positive drug group myocardial pathological morphology；
Fig. 7 indicates the case where expression of CRP, IL-6 and TNF-α in the experiment three of embodiment 4；
Fig. 8 shows TXA in the experiment three of embodiment 42And PGI2The case where expression；
NF-KB immunohistochemistry positive expression in different experiments group rat myocardial cell in the experiment three of Fig. 9 expression embodiment 4 The case where；Wherein Fig. 9-a is sham-operation group；Fig. 9-is model group；Fig. 9-c is low dose group；Fig. 9-d is middle dose group；Fig. 9-e For high dose group；9-f is positive drug group.
Specific embodiment
The present invention provides a kind of Chinese medicine composition for treating acute coronary syndrome, the raw material including following parts by weight: 20~50 parts of honeysuckle (dry flower of caprifoliaceae plant honeysuckle Lonicera japonica Thunb. or with the flower just opened), Root of kirilow rhodiola (the dry root and root of crassulaceae plants flower red-spotted stonecrop Rhodiola crenulata (Hook.f.et Thoms.) H.Ohba Stem) 20~50 parts, leech (Hirudinidae animal leech Whitmania pigra Whitman, leech Hirudo nipponica Whitman's or whitmania acranulata Whitman Whitmania acranulata Whitman is dry all) 5~20 parts, Snakegourd Fruit (plant by Curcurbitaceae Object snakegourd Trichosanthes kiriloxvii Maxim. or trichosanthes rosthornii Harms Trichosanthes rosthornii Harms Dry mature fruit) 10~50 parts, radix scrophulariae (scrophulariaceae radix scrophulariae Scrophularia ningpoensis Hemsl.'s Dry root) 10~30 parts, Radix Angelicae Sinensis (dry root of umbelliferae angelica Angelica sinensis (Oliv.) Diels) 20~ 40 parts, 10~30 parts of rhizoma nardostachyos (the drying root and rhizome of patrinia herb rhizoma nardostachyos Nardostachys jatamansi DC.), pellet Join 20~40 parts of (dry root and rhizome of Lamiaceae plant Radix Salviae Miltiorrhizae Salvia miltiorrhiza Bge.) and Radix Glycyrrhizae (pulse family plant Object Radix Glycyrrhizae Glycyrrhiza uralensis Fisch., swollen fruit Radix Glycyrrhiza inflata Bat. or glycyrrhiza glabra The dry root and rhizome of Glycyrrhiza glabra L.) 5~10 parts.
Chinese medicine composition of the present invention includes 20~50 parts of honeysuckle, preferably 30~40 parts, more preferably 32~38 Part；Organic acid reacts rapidly with peroxy radical in the honeysuckle, protects vascular endothelial cell damage, avoids blood platelet living Change, thus the aggregation of blocking platelet.
Chinese medicine composition of the present invention includes 10~30 parts of radix scrophulariae, preferably 15~25 parts, more preferably 18~22 parts； The radix scrophulariae can inhibit myocardial fibrosis, delay the development process of left ventricular remodeling；And radix scrophulariae alcohol medicinal extract aqueous solution can obviously increase Add coronary blood flow；There is similar beta-blocker effect in radix scrophulariae, have slight inhibiting effect to heart rate, cardiac contractile force；It removes Except this, radix scrophulariae also has the function of platelet aggregation-against.
Chinese medicine composition of the present invention includes 20~40 parts of Radix Angelicae Sinensis, preferably 25~35 parts, more preferably 28~32 parts； The Radix Angelicae Sinensis can increase coronary blood flow, reduce heart infarction range, enhance anti-anoxia ability, inhibit platelet aggregation.
Chinese medicine composition of the present invention includes 20~40 parts of Radix Salviae Miltiorrhizae, preferably 25~35 parts, more preferably 28~32 parts, The Radix Salviae Miltiorrhizae has coronary artery dilator and peripheral blood vessel, increases coronary blood flow, and reducing heart rate mitigates myocardial ischemic injury Degree；It can promote fibrinolysis and have anticoagulation；In addition, Radix Salviae Miltiorrhizae has the free radical for understanding ishemic part, enhance the heart Myotility improves hypoxia-bearing capability, reduces the effect of myocardial infarct size.
Chinese medicine composition of the present invention includes 5~10 parts of Radix Glycyrrhizae, preferably 6~9 parts, more preferably 7~8 parts；It is described Glycyrrhizin in licorice can reduce the content of Blood Cholesterol and free fatty acid, reduce the heap of triglycerides Product；And the extract of Radix Glycyrrhizae can extend blood plate thrombus and form time, reinforcing fiber protein dissolution activity.
Chinese medicine composition of the present invention includes 20~50 parts of root of kirilow rhodiola, preferably 30~40 parts, more preferably 32~38 Part；The effect of the root of kirilow rhodiola is qi and activate blood circulation, promotes blood circulation and relievings asthma.
Chinese medicine composition of the present invention includes 5~20 parts of leech, preferably 10~15 parts, more preferably 12~14 parts； The effect of the leech is blood-breaking promoting menstruation, by stagnation resolvation disease.
Heretofore described Chinese medicine composition includes 10~50 parts of Snakegourd Fruit, preferably 20~40 parts, and more preferable 25~35 parts； The effect of the Snakegourd Fruit is clearing away heat and eliminating phlegm, relieving stuffiness of the chest by dispersing aggregation of pathogens, moisturizing dryness and lubricating intestine.
Heretofore described Chinese medicine composition includes 10~30 parts of rhizoma nardostachyos, preferably 15~25 parts, and more preferable 18~22 parts； The effect of the rhizoma nardostachyos is regulating qi-flowing for relieving pain, opens strongly fragrant be amusing.
In the present invention, the Chinese medicine composition has the effect of dispelling toxins and dredging collaterals, blood stasis dispersing and fresh blood promoting.
The present invention provides the preparation methods of the Chinese medicine composition described in above scheme, comprising the following steps:
1) Radix Salviae Miltiorrhizae being mixed with ethanol water, carries out refluxing extraction, filtering collects filtrate concentration, obtains the first thick paste, Collection filter residue is first dregs of a decoction；
2) honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos are mixed with water, carry out the first water extraction, collection obtain volatile oil, extracting solution and Second dregs of a decoction；
3) by step 1) first dregs of a decoction, step 2) second dregs of a decoction and root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae, Radix Glycyrrhizae It is mixed with water, carries out the second water extraction, filtering, concentrated filtrate obtain molten cream；
4) it stands, extracts after mixing step 3) the molten cream with ethanol water, filtering, concentrated filtrate obtain Second thick paste；
5) quiet after mixing step 1) first thick paste, step 4) second thick paste and the step 2) extracting solution It sets, filtering, dried filtrate, dried object crush to obtain medicinal powder；
6) by volatile oil beta-cyclodextrin inclusion compound described in step 2), inclusion compound is obtained；
7) inclusion compound described in step 6) is mixed with medicinal powder described in step 5), obtains Chinese medicine composition.
Before preparation method of the present invention, preferably to raw material honeysuckle, root of kirilow rhodiola, leech, Snakegourd Fruit, radix scrophulariae, Radix Angelicae Sinensis, Rhizoma nardostachyos, Radix Salviae Miltiorrhizae and Radix Glycyrrhizae are selected, are cleaned, eluted and are dried pretreatment；It is described to select, clean, elute and dry pretreatment It is carried out using this field conventional means and standard.
The present invention mixes after pre-processing to Chinese medicine material, by Radix Salviae Miltiorrhizae with ethanol water, carries out refluxing extraction, mistake Filtrate concentration is collected in filter, obtains the first thick paste, and collection filter residue is first dregs of a decoction；The volume hundred of ethyl alcohol in the ethanol water Divide than being 80%~95%, preferably 85%~90%；The mass ratio of the Radix Salviae Miltiorrhizae and ethanol water is 1:(7~9), preferably For 1:8；
In the present invention, the temperature of the refluxing extraction is 60~80 DEG C, preferably 65~75 DEG C, more preferably 70 DEG C；Institute The number for stating refluxing extraction is 2~4 times, preferably 3 times；The time of each refluxing extraction is preferably 1~3h, more preferably 2h；The mode of the filtering is preferably membrane filtration；The aperture of the filter membrane is preferably 80~120 mesh, more preferably 100 mesh； The mode of the concentration is preferably concentrated under reduced pressure；The pressure of the reduced pressure is preferably 0.08~0.09Mpa；The decompression is dense The temperature of contracting is preferably 45~55 DEG C, and more preferably 50 DEG C；The time of the reduced pressure is preferably 2~5h, more preferably 3~ 4h；The relative density of first thick paste is preferably 1.10~1.15.
In the present invention, honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos are mixed with water, carry out the first water extraction, collection obtain volatile oil, Extracting solution and second dregs of a decoction；The mass ratio of the honeysuckle, the gross mass of Radix Angelicae Sinensis and rhizoma nardostachyos and water is 1:(9~11), preferably 1:10；The time that first water extracts is preferably 5~7h, more preferably 6h；The number that first water extracts is preferably 1 It is secondary；The temperature that first water extracts is preferably 80~100 DEG C, and more preferably 90 DEG C；The equipment that first water extracts is preferred For Chinese medicine extracting tank.
In the present invention after obtaining first dregs of a decoction and second dregs of a decoction, by first dregs of a decoction, second dregs of a decoction and root of kirilow rhodiola, Snakegourd Fruit, Leech, radix scrophulariae, Radix Glycyrrhizae and water mixing, carry out the second water extraction, and filtering, concentrated filtrate obtain molten cream；First dregs of a decoction, The mass ratio of two dregs of a decoction, root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae and the gross mass of Radix Glycyrrhizae and water is preferably 1:(5~7), preferably 1: 6。
In the present invention, the equipment that second water extracts is preferably Chinese medicine extracting tank；The temperature that second water extracts is 80~100 DEG C, preferably 85~95 DEG C, more preferably 90 DEG C；The number that second water extracts is 2~4 times, preferably 3 times； The time extracted every time is 1~3h, preferably 2h；The mode of the filtering is preferably membrane filtration；The aperture of the filter membrane is excellent It is selected as 80~120 mesh, more preferably 100 mesh；The mode of the concentration is preferably concentrated under reduced pressure；The pressure of the reduced pressure is excellent It is selected as 0.08~0.09Mpa；The temperature of the reduced pressure is preferably 45~55 DEG C, and more preferably 50 DEG C；The reduced pressure Time be preferably 2~5h, more preferably 3~4h；The relative density of the molten cream is preferably 1.10~1.15.
The present invention mixes after obtaining molten cream, by molten cream with ethanol water, extracts, and filtering, concentrated filtrate obtain To the second thick paste；The percent by volume of ethyl alcohol is preferably 75%~85% in the ethanol water, and more preferably 80%；It is described The mass ratio of ethanol water and molten cream is preferably 5~7:3~5, more preferably 6:4.
In the present invention, the temperature of the standing is preferably 1~5 DEG C, and more preferably 4 DEG C；The time of the standing is preferably 18~30h, more preferably for 24 hours；The mode of the filtering is preferably membrane filtration；The aperture of the filter membrane is preferably 80~120 Mesh, more preferably 100 mesh；The mode of the concentration is preferably concentrated under reduced pressure；The pressure of the reduced pressure is preferably 0.08~ 0.09Mpa；The temperature of the reduced pressure is preferably 45~55 DEG C, and more preferably 50 DEG C；The time of the reduced pressure is preferred For 2~5h, more preferably 3~4h；The relative density of second thick paste is preferably 1.10~1.15.
The present invention is after obtaining the first thick paste, the second thick paste and extracting solution, by the first thick paste, the second thick paste and extracting solution It is stood after mixing, filtering, dried filtrate, dried object crush to obtain medicinal powder；The temperature of the standing be 1~5 DEG C, preferably 4 ℃；The time of the standing is 18~30h, preferably for 24 hours；The mode of the drying is preferably dried under reduced pressure；The decompression is dry Dry temperature is preferably 60~80 DEG C, and more preferably 70 DEG C；The time being dried under reduced pressure is preferably 48~96h, more preferably 72h；The partial size of the medicinal powder is 80~100 mesh, preferably 90 mesh.
The present invention, by the volatile oil beta-cyclodextrin inclusion compound, obtains inclusion compound after obtaining volatile oil.
The present invention provides the preparation methods of the inclusion compound, comprising the following steps:
A) volatile oil is mixed with ethanol solution, obtains the ethanol solution of volatile oil；
B) ethanol solution, beta-cyclodextrin and water of the step a) volatile oil are mixed, obtains beta-cyclodextrin saturation Aqueous solution carries out inclusion processing, and refrigeration filters, and washing filter residue is dried to obtain inclusion compound.
In the present invention, the volatile oil is mixed with ethanol solution, obtains the ethanol solution of volatile oil；Institute The volume ratio for stating volatile oil and ethanol solution is preferably 4~6:4~6, more preferably 1:1.
The present invention is after obtaining the ethanol solution of volatile oil, by the volatile oil ethanol solution, beta-cyclodextrin It is mixed with water, obtains beta-cyclodextrin saturated aqueous solution, carry out inclusion processing, refrigeration filters, and washing filter residue is dried to obtain inclusion Object；The volume mass ratio of volatile oil and cyclodextrin in the volatile oil ethanol solution is preferably 1mL:6~10g, More preferably 1mL:7~9g；The equipment of the inclusion processing is preferably magnetic stirring apparatus；It is described inclusion processing temperature be preferably 40~60 DEG C, more preferably 50 DEG C；The time of the inclusion processing is preferably 1~3h, more preferably 2h；The chilling treatment Temperature is preferably 1~5 DEG C, and more preferably 4 DEG C；The time of the chilling treatment is preferably 18~30h, more preferably for 24 hours；It is described Suction filtration mode is preferably that filter paper filters；The solvent of the washing is preferably petroleum ether；The anhydrous second of the petroleum ether and volatile oil The volume ratio of alcoholic solution is preferably 1:(9~11), more preferably 1:10；The temperature of the petroleum ether is preferably 60~90 DEG C, more Preferably 79~80 DEG C；The number of the washing is preferably 2~4 times, and more preferably 3 times；The equipment of the drying is preferably true Empty drying box；The temperature of the drying is preferably 40~60 DEG C, and more preferably 50 DEG C；The time of the drying is preferably 3~5h, More preferably 4h.
The present invention mixes after obtaining inclusion compound and medicinal powder, by inclusion compound with medicinal powder, obtains Chinese medicine composition.
The present invention provides the preparation methods of the Chinese herbal granules comprising Chinese medicine composition described in above scheme, and step is such as Under: after Chinese medicine composition, dextrin and ethanol water are mixed, is pelletized, is dried to obtain Chinese herbal granules；The ethanol water Solution is wetting agent.Wherein, the preparation method flow chart of the Chinese herbal granules is shown in Fig. 1.
In the present invention, the percent by volume of ethyl alcohol is preferably 70%~80% in the ethanol water, more preferably 75%；The mass ratio of the Chinese medicine composition and dextrin is preferably 1:0.5~2, more preferably 1:1~1.5；The temperature of the drying Preferably 70~90 DEG C, more preferably 80 DEG C of degree；The time of the drying is preferably 12~36h, more preferably for 24 hours；In described The water content of medicine granule is preferably≤8%；The partial size of the Chinese herbal granules is preferably 10~80 mesh, more preferably 30~50 Mesh.
Below with reference to embodiment to a kind of Chinese medicine composition for treating acute coronary syndrome provided by the invention and its system Preparation Method is described in detail, but they cannot be interpreted as limiting the scope of the present invention.
A kind of Chinese herbal granules for treating acute coronary syndrome of embodiment 1
1. pretreatment: extracting honeysuckle 200g, root of kirilow rhodiola 200g, leech 50g, Snakegourd Fruit 100g, radix scrophulariae 100g, Radix Angelicae Sinensis 200g, Rhizoma nardostachyos 100g, Radix Salviae Miltiorrhizae 200g and Radix Glycyrrhizae 50g, and each taste medicine is chosen to miscellaneous, elution, drying；
2. taking pretreated raw material Radix Salviae Miltiorrhizae, adding the percentage by volume of 8 times of Radix Salviae Miltiorrhizae quality is 80% ethanol water, is adopted It is extracted, 60 DEG C, is extracted 3 times, each 1h, 100 mesh membrane filtrations with Effect of Alcohol Extracting Procedureto Total, the dregs of a decoction are spare；Merging filtrate obtains second Alcohol extract and first dregs of a decoction；
3. (pressure 0.08Mpa) 2h is concentrated under reduced pressure in ethanol extract under the conditions of 50 DEG C in step 2, relative density is obtained 1.10 the first thick paste；
4. taking pretreated raw material honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos, add 10 times of honeysuckles, Radix Angelicae Sinensis and rhizoma nardostachyos quality Water is extracted volatile oil 1 time, time 6h using water extraction process, and extraction obtains volatile oil；Filtering, obtains extracting solution and the second medicine Slag.
5. first dregs of a decoction, second dregs of a decoction and pretreated raw material root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae and Radix Glycyrrhizae are mixed, The water of 6 times of said mixture quality is added to extract 80 DEG C, 3 times, each 1h, 100 mesh membrane filtrations are concentrated under the conditions of 50 DEG C of filtrate Obtain the molten cream that relative density is 1.10；
6. the ethyl alcohol that cream molten in step 5 is added 80% is stirred evenly until ethyl alcohol quality accounts for the 60% of total weight, 4 DEG C quiet It sets and obtains ethanol for 24 hours；Said mixture is filtered using 100 mesh filter membranes, being recovered under reduced pressure and being concentrated to get relative density is 1.10 The second thick paste；
7. the first thick paste in step 3 is poured into the second thick paste of step 6, it is uniformly mixed, the extraction in step 4 is added Liquid, mixing, 4 DEG C stand for 24 hours, filtering, 70 DEG C be dried under reduced pressure, crushing to obtain partial size for dried object is 80 mesh medicinal powder；
8. by volatile oil beta-cyclodextrin inclusion compound in step 4: taking (50% volatilization of 50% volatile oil ethanol solution of 1mL Oily ethanol solution is that will volatilize oil and dehydrated alcohol with solution that volume ratio is that 1:1 is mixed to get), according to volatile oil It is configured in saturated aqueous solution, is placed in magnetic stirring apparatus for the amount of 1mL:6g with cyclodextrin envelope-bulk to weight ratio, setting inclusion temperature Degree is 40 DEG C, constant temperature stirs 1h, is cooled to room temperature, and for 24 hours, filter paper filters for 4 DEG C of refrigerations, discards filtrate, 60 DEG C of petroleum ethers of filter residue Washing 3 times, each 10mL, discards petroleum ether washing lotion, and filter residue sets 4h in 50 DEG C of vacuum ovens, takes out, obtains inclusion；
9, inclusion is mixed with step 7 traditional Chinese medicine powder, according to weight ratio, 75% ethyl alcohol of mixed powder and dextrin 1:0.5 Aqueous solution is wetting agent, and granulation, granular size is 80 mesh, and 80 DEG C of air drying 12h obtain Chinese herbal granules.
A kind of Chinese herbal granules for treating acute coronary syndrome of embodiment 2
1. pretreatment: extracting honeysuckle 500g, root of kirilow rhodiola 500g, leech 200g, Snakegourd Fruit 500g, radix scrophulariae 300g, Radix Angelicae Sinensis 400g, Rhizoma nardostachyos 300g, Radix Salviae Miltiorrhizae 400g and Radix Glycyrrhizae 100g, and each taste medicine will be chosen to miscellaneous, elution, drying；
2. taking pretreated raw material Radix Salviae Miltiorrhizae, adding the percentage by volume of 9 times of Radix Salviae Miltiorrhizae quality is 85% ethanol water, is adopted It is extracted, 80 DEG C, is extracted 3 times, each 2h, 100 mesh membrane filtrations with Effect of Alcohol Extracting Procedureto Total, the dregs of a decoction are spare；Merging filtrate obtains second Alcohol extract and first dregs of a decoction；
3. 2h is concentrated under reduced pressure under conditions of 50 DEG C, pressure 0.09Mpa in ethanol extract in step 2, relative density is obtained For 1.15 the first thick paste；
4. taking pretreated raw material honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos, add 10 times of honeysuckles, Radix Angelicae Sinensis and rhizoma nardostachyos quality Water is extracted volatile oil 1 time, time 6h using water extraction process, and extraction obtains volatile oil；Filtering, obtains extracting solution and the second medicine Slag.
5. first dregs of a decoction, second dregs of a decoction and pretreated raw material root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae and Radix Glycyrrhizae are mixed, The water of 6 times of said mixture quality is added to extract 90 DEG C, 3 times, each 2h, 100 mesh membrane filtrations are concentrated under the conditions of 50 DEG C of filtrate Obtain the molten cream that relative density is 1.15；
6. the ethyl alcohol that cream molten in step 5 is added 80% is stirred evenly until ethyl alcohol quality accounts for the 60% of total weight, 4 DEG C quiet It sets and obtains ethanol for 24 hours；Said mixture is filtered using 100 mesh filter membranes, being recovered under reduced pressure and being concentrated to get relative density is 1.10 The second thick paste；
7. the first thick paste in step 3 is poured into the second thick paste of step 6, it is uniformly mixed, the extraction in step 4 is added Liquid, mixing, 4 DEG C stand for 24 hours, filtering, 70 DEG C be dried under reduced pressure, crushing to obtain partial size for dried object is 80 mesh medicinal powder；
8. by volatile oil beta-cyclodextrin inclusion compound in step 4: taking (50% volatilization of 50% volatile oil ethanol solution of 1mL Oily ethanol solution is that will volatilize oil and dehydrated alcohol with solution that volume ratio is that 1:1 is mixed to get), according to volatile oil With cyclodextrin volume mass than being configured in saturated aqueous solution for the amount of 1mL:6g, it is placed in magnetic stirring apparatus, setting inclusion temperature Degree is 40 DEG C, constant temperature stirs 1h, is cooled to room temperature, and for 24 hours, filter paper filters for 4 DEG C of refrigerations, discards filtrate, 60 DEG C of petroleum ethers of filter residue Washing 3 times, each 10mL, discards petroleum ether washing lotion, and filter residue sets 4h in 50 DEG C of vacuum ovens, takes out, obtains inclusion；
9, inclusion is mixed with step 7 traditional Chinese medicine powder, according to mixed powder and (w:w) ratio of dextrin 1:0.5,75% ethyl alcohol Aqueous solution is wetting agent, and granulation, granular size is 80 mesh, and 80 DEG C of air drying 36h obtain Chinese herbal granules.
A kind of Chinese herbal granules for treating acute coronary syndrome of embodiment 3
1. pretreatment: extracting honeysuckle 300g, root of kirilow rhodiola 400g, leech 100g, Snakegourd Fruit 300g, radix scrophulariae 200g, Radix Angelicae Sinensis 300g, Rhizoma nardostachyos 200g, Radix Salviae Miltiorrhizae 300g and Radix Glycyrrhizae 75g, and each taste medicine will be chosen to miscellaneous, elution, drying；
2. taking pretreated raw material Radix Salviae Miltiorrhizae, adding the percentage by volume of 10 times of Radix Salviae Miltiorrhizae quality is 90% ethanol water, It is extracted, 80 DEG C, is extracted 3 times, each 3h, 100 mesh membrane filtrations using Effect of Alcohol Extracting Procedureto Total, the dregs of a decoction are spare；Merging filtrate obtains Ethanol extract and first dregs of a decoction；
3. 2h is concentrated under reduced pressure under conditions of 50 DEG C, pressure 0.09Mpa in ethanol extract in step 2, relative density is obtained For 1.15 the first thick paste；
4. taking pretreated raw material honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos, add 10 times of honeysuckles, Radix Angelicae Sinensis and rhizoma nardostachyos quality Water is extracted volatile oil 1 time, time 6h using water extraction process, and extraction obtains volatile oil；Filtering, obtains extracting solution and the second medicine Slag.
5. first dregs of a decoction, second dregs of a decoction and pretreated raw material root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae and Radix Glycyrrhizae are mixed, The water of 6 times of said mixture quality is added to extract 100 DEG C, 3 times, each 3h, 100 mesh membrane filtrations are dense under the conditions of 50 DEG C of filtrate Contracting obtains the molten cream that relative density is 1.15；
6. the ethyl alcohol that cream molten in step 5 is added 80% is stirred evenly until ethyl alcohol quality accounts for the 60% of total weight, 4 DEG C quiet It sets and obtains ethanol for 24 hours；Said mixture is filtered using 100 mesh filter membranes, being recovered under reduced pressure and being concentrated to get relative density is 1.10 The second thick paste；
7. the first thick paste in step 3 is poured into the second thick paste of step 6, it is uniformly mixed, the extraction in step 4 is added Liquid, mixing, 4 DEG C stand for 24 hours, filtering, 70 DEG C be dried under reduced pressure, crushing to obtain partial size for dried object is 80 mesh medicinal powder；
8. by volatile oil beta-cyclodextrin inclusion compound in step 4: taking (50% volatilization of 50% volatile oil ethanol solution of 1mL Oily ethanol solution is that will volatilize oil and dehydrated alcohol with solution that volume ratio is that 1:1 is mixed to get), according to volatile oil With cyclodextrin volume mass than being configured in saturated aqueous solution for the amount of 1mL:6g, it is placed in magnetic stirring apparatus, setting inclusion temperature Degree is 40 DEG C, constant temperature stirs 1h, is cooled to room temperature, and for 24 hours, filter paper filters for 4 DEG C of refrigerations, discards filtrate, 60 DEG C of petroleum ethers of filter residue Washing 3 times, each 10mL, discards petroleum ether washing lotion, and filter residue sets 4h in 50 DEG C of vacuum ovens, takes out, obtains inclusion；
9, inclusion is mixed with step 7 traditional Chinese medicine powder, according to mixed powder and (w:w) ratio of dextrin 1:0.5,75% ethyl alcohol Aqueous solution is wetting agent, and granulation, granular size is 80 mesh, and 80 DEG C of air drying 36h obtain Chinese herbal granules.
The Chinese herbal granules of 4 embodiment 3 of embodiment are to Non-ST Elevation Acute type Rat of Myocardial Infarction caused by coronary artery ligation Influence
A Model Rats with Acute Myocardial Ischemia is tested to establish and evaluate
Wistar rat 40, half male and half female, weight (230 ± 10) g, by the zoopery of preclinical medicine institute, Jilin University The heart provides.Credit number: FCXK (Ji) -2011-0004.Rat is fed in standard animal chambers, and rat adaptable fed 1 week starts Experiment.By above-mentioned 40 Wistar rats, it is divided into sham-operation group, model group by random digits table, every group 10, male and female are each Half.
1 establishes Model Rats with Acute Myocardial Ischemia, the method is as follows:
1) preoperative fasting in 24 hours, gives suitable water；2) etherization layback position is fixed on operating table；3) center is shaved Hair, does purse string suture with No. 10 line, with scalpel midsection, removes intercostal muscle with haemostatic clamp, 3~4 intercostal is opened from left side Chest, exposure heart；4) coronary artery left anterior descending branch is found out between pulmonary conus and atrium sinistrum, sham-operation group, which only threads, not to ligature outside, Model group ligatures left anterior descending branch with No. 0 line immediately；It sends heart back to thoracic cavity, tightens pocket suture rapidly after squeezing out chest chamber air Close thoracic cavity.
Indices are acquired after 2 modelings 24 hours
2.1 electrocardiographic recorder
Coronary ligation is preoperative, and standard limb II lead electrocardiogram under normal condition is recorded after etherization.By three aciculiforms Electrode by specified position (it is yellow --- right upper extremity, it is black --- right lower extremity, it is red --- left lower extremity) under insertion rat four limbs cortex.It is postoperative 24 hours, after 20% urethane (1.0g/kg) intraperitoneal anesthesia, record standard limbs II lead electrocardiogram again.Calculate each mark ST sections of variation millivolt numbers of measuring point.
The measurement of 2.2 myocardial infarct sizes
Heart is taken out, with normal saline flushing, filter paper is blotted, and by ventricle crosscutting 5, sets nitroblue tetrazolium chloride (NB- T it) in phosphate buffer, is taken out after 37 DEG C of water bath with thermostatic control dyeing completely.After digital camera is taken pictures, analyzed using morphological images System software 1.0 measures relevant range area, and infarct size (%)=left ventricular tissues slice infarct size/left ventricular tissues are cut The piece gross area × 100%.
The detection of 2.3 serum enzymes
Abdominal aorta blood sampling 5ml, is placed in test tube, and with 3000rpm/min, after being centrifuged 10min, Aspirate supernatant is made, and puts It is placed in ultra low temperature freezer, -20 DEG C of preservations, glutamic-oxalacetic transaminease (AST) to be detected, lactic dehydrogenase (LDH), creatine enzyme isoenzyme (CK-MB), Troponin I (cTn-I).Every takes 0.5ml serum, 7600 automatic clinical chemistry analyzer of Yu Li detect AST, LDH,CK-MB.Every takes 10ul serum, using double crush syndrome Enzyme-linked Immunosorbent Assay method, measures cTn-I.In strict accordance with Kit operating method is detected.
3 statistical analysis
Statistical procedures are carried out using SPSS16.0 software.Measurement data is with mean ± standard deviationIt indicates, group Between mean comparison use one-way analysis of variance, P < 0.05 be difference it is statistically significant.Statistical result is as follows:
1) ECG Change: 24 hours after model group modeling, ∑ ST variation is obvious (P < 0.05) compared with sham-operation group, referring to Table 2.
2) myocardial infarct size compares: (P < 0.05) is dramatically increased with sham-operation group comparison model group myocardial infarct size, referring to table 2.
Before 2 modeling of table and ECG ST section changes after modeling and myocardial infarct size measures
* the P=0.000 compared with model group；# P=0.000 compared with model group
3) serum enzyme testing result: model group AST, LDH, CK-MB, TN-I concentration is higher than sham-operation group, has statistics It learns meaning (P < 0.05).It is shown in Table 1-2 and Fig. 2, Fig. 2 is myocardium enzyme and the case where Applications of Cardiac Markers is expressed.
The case where table 1-2 myocardium enzyme and Applications of Cardiac Markers are expressed
* the P=0.008 compared with sham-operation group；* P=0.008 compared with sham-operation group；
# P=0.001 compared with sham-operation group；@P=0.000 compared with sham-operation group.
From ECG ST section variation, myocardial infarct size, serum enzyme comparison result before and after two groups of modelings, this experiment is absolutely proved It is successfully, reproduced out the animal model of acute myocardial ischemia.
Test the confirmatory experiment of the protective effect of the Chinese herbal granules of two embodiments 3 to Acute Myocardial Ischemia Rats
Wistar rat 120, half male and half female, weight (230 ± 10) g, by the zoopery of preclinical medicine institute, Jilin University Center provides.Credit number: FCXK (Ji) -2011-0004.Rat is fed in standard animal chambers, opens within rat adaptable fed 1 week Begin to test.Experimental drug: the Chinese herbal granules and Tongxinluo medicinal capsule of embodiment 3 (are mentioned by Yiling Pharmaceutical Co., Ltd, Shijiazhuang For batch number: 110919).
1. 120 Wistar rats of modeling are divided into sham-operation group, model group, Chinese herbal granules according to random digits table Large, medium and small dosage group, Tongxinluo group.Sham-operation group and model group stomach-filling sterile purified water 2ml/d are only；Chinese herbal granules are big, In, small dose group distinguish stomach-filling 1.528g/kg, 3.055g/kg, 6.11g/kg；Tongxinluo group stomach-filling 0.24g/kg.Continuous gavage Modeling is carried out according to the modeling method in above-mentioned experiment one after 14 days.Sham-operation group, which only threads, not to ligature outside, remaining each group is with 0 Number line ligatures left anterior descending branch immediately.
2. observation index observation index is；ECG ST section variation before and after modeling；Myocardial infarct size measurement；Serum enzyme (AST,LDH,CK-MB,cTnI)；Cardiac morphology variation.In addition to myocardial morphology variation, remaining materials is with detection method with experiment One.
Myocardial pathological morphology variation materials and observation method: it takes out heart and takes at apex of the heart 1cm with a thickness of 0.5cm Cross-section cardiac muscular tissue's block, be soaked in 10% formalin solution 24 hours, routine paraffin wax embedding, slice, to HE dye.
HE staining procedure:
(1) 95% ethyl alcohol I → 3min of 10min dimethylbenzene I → 10min dimethylbenzene II → 3min, 95% ethyl alcohol II → 1min80% ethyl alcohol → 1min aquation
(2) haematoxylin dyeing 5min, tap water rinse
(3) 1% acidic alcohols break up 30s
(4) tap water impregnates 10min
(5) Yihong liquid 1min is set, originally who is rinsed
95% ethyl alcohol II → 2min dimethylbenzene I → 2min diformazan of (6) 80% ethyl alcohol 10s → 3min95% ethyl alcohol I → 3min Benzene II → resinene mounting
(7) it takes pictures
3. statistical analysis carries out statistical procedures using SPSS16.0 software.Measurement data is with mean ± standard deviationIt indicates, the comparison of mean uses one-way analysis of variance between group, and P < 0.05 is that difference is statistically significant.As a result it shows Show:
1) ECG ST section variation is compared before and after modeling: between each group before modeling with for 24 hours, ∑ ST changes comparison result after modeling Illustrate, compared with model group, treatment group ∑ ST changes unobvious (P < 0.05), and wherein high dose group and Tongxinluo group effect are bright It is aobvious.3, Fig. 3 are shown in Table, wherein the situation of change before Fig. 3 expression modeling with electrocardiogram after modeling.
Before 3 modeling of table and modeling after electrocardiogram situation of change
* P value is followed successively by 0.001,0.012,0.034,0.018 compared with sham-operation group
△ P value compared with model group is followed successively by 0.001,0.000,0.000,0.000
2) myocardial infarct size compares: between each group myocardial infarct size comparison result show low dose group compared with model group, heart infarction face Product, which has no, significantly reduces (P > 0.05)；Remaining treatment group substantially reduces myocardial infarct size (P < 0.05) compared with model group, especially with high agent Amount group is obvious；High dose group and Tongxinluo group reduction myocardial infarct size effect are much the same.It is shown in Table 4, Fig. 4, wherein Fig. 4 indicates different The myocardial infarct size of experimental group.
Influence of the table 4 to myocardial infarct size
△ P value compared with model group is followed successively by 0.330,0.001,0.001,0.001
3) expression of myocardium enzyme and Applications of Cardiac Markers
The experimental results showed that high dose group and Tongxinluo group significantly reduce AST, LDH, CK-MB, cTn-I compared with model group Concentration (P < 0.05)；Low dose group can reduce concentration (the P < of CK-MB, cTnI compared with middle dose group is compared with model group , but AST, LDH concentration no significant difference (P > 0.05) 0.05).It is shown in Table 5, Fig. 5, Fig. 5 indicates myocardium enzyme and Applications of Cardiac Markers table Up to the case where.
The influence that table 5 expresses myocardium enzyme and Applications of Cardiac Markers
* the P=0.008 compared with sham-operation group；△ P value compared with model group is followed successively by 0.290,0.076,0.039, 0.045；
* P=0.008 compared with sham-operation group；△ △ P value compared with model group is followed successively by 0.348,0.314,0.011, 0.003；
# P=0.001 compared with sham-operation group；@P value compared with model group is followed successively by 0.017,0.010,0.010, 0.005；
## P=0.000 compared with sham-operation group；@@P value compared with model group is followed successively by 0.000,0.000,0.000, 0.000；
HE dyeing, microscope observe myocardial pathological morphology under (× 100), as a result see that Fig. 6, Fig. 6 indicate different experiments group Myocardial pathological morphology, wherein Fig. 6-a is sham-operation group myocardial pathological morphology；Fig. 6-b is model group myocardial pathological morphology；Fig. 6-c For low dose group myocardial pathological morphology；Fig. 6-d is middle dose group myocardial pathological morphology；Fig. 6-e is high dose group myocardium markers shape State；6-f is positive drug group myocardial pathological morphology.As the result is shown: sham-operation group: the cardiac morphology of rat heart is normal, and structure is clear Clear, muscle fibre band, intercalated disc and nuclear targeting are clear, assemble hyperplasia phenomenon without fibrocyte between muscle fibre；Model group rats In no configuration state within the scope of cardiac infarction area, cardiac muscle cell's karyopycnosis, fragmentation or dissolution disappear, and part cardiac muscle is in stove shape and piece Shape necrosis, cardiac muscle fibre disorder, fracture are obvious, myoplasm cohesion, unclear transverse striation of muscle fiber or disappearance, the visible red blood cell of iuntercellular, with more Unrestrained neutral polymorphonuclear cell infiltration, and have the denaturation of part cardiac muscle fibre sample.Infarct does not involve ventricle wall holostrome, periinfarct The substantially same normal myocardium of tissue morphology in area and separate infarcted region；Each treatment group's rat heart muscle degree of necrosis mitigates, inflammatory cell Infiltration is reduced, and degree of myocardial fibrosis mitigates, the most significant with high-dose therapy group.The myocardium cell injuring degree of each treatment group It is light compared with model group, it was demonstrated that the Chinese herbal granules of embodiment 3 have the function of mitigating myocardial ischemia in rats damage.
Experimental result is shown: cardiac muscle cell's karyopycnosis, broken in no configuration state within the scope of model group rats cardiac infarction area Disappearance is split or dissolves, for part cardiac muscle in stove shape and sheet necrosis, cardiac muscle fibre disorder, fracture are obvious, myoplasm cohesion, unclear transverse striation of muscle fiber Or disappear, the visible red blood cell of iuntercellular with the neutral polymorphonuclear cell infiltration diffused, and has the change of part cardiac muscle fibre sample Property.And the high, medium and low dosage group of Chinese herbal granules and positive controls rat heart muscle degree of necrosis of embodiment 3 mitigate, inflammatory is thin Born of the same parents, which infiltrate, to be reduced, and degree of myocardial fibrosis mitigates, the most significant with high-dose therapy group.Illustrate the Chinese herbal granules of embodiment 3 With mitigation ischemic myocardium damaging action.
By above-mentioned experimental result it is found that the Chinese herbal granules of the embodiment of the present invention 3 can significantly reduce ST sections of variations, significantly Rat myocardial infarct size is reduced, and AST, LDH, CK-MB and cTnI activity can be substantially reduced, mitigates myocardial ischemia in rats damage Wound.
Test research of the Chinese herbal granules of three embodiment of the present invention 3 to Acute Myocardial Ischemia Rats inflammatory mechanisms
Wistar rat 120, half male and half female, weight (230 ± 10) g, by the zoopery of preclinical medicine institute, Jilin University Center provides.Credit number: FCXK (Ji) -2011-0004.Rat is fed in standard animal chambers, opens within rat adaptable fed 1 week Begin to test.
1. experimental drug is the same as experiment two.Grouping, administration and modeling method are the same as experiment two.Observation index are as follows: C in rat blood serum Reactive protein, thromboxane, prostacyclin, interleukin-6 concentration；NF-KBp65 is expressed in cardiac muscle cell.
2. detection method
1) c reactive protein, thromboxane, prostacyclin and interleukin-6 are all made of double crush syndrome Enzyme-linked Immunosorbent Assay Method is detected in strict accordance with kit operating method.
2) detection NF-KBp65 expression uses SABC Immunohistochemical Method.Specific steps are as follows:
1. after paraffin specimen routine 4um serial section, being placed in the anticreep fragmentation handled through poly-D-lysine, being put in 60 DEG C Oven is stayed overnight.
2. dewaxing and aquation: histotomy is placed in dimethylbenzene I, II each 15min → dehydrated alcohol I5min that dewaxes → anhydrous The ethyl alcohol of ethyl alcohol II3min → 95% ethyl alcohol of 3min → 80% ethyl alcohol of 3min → 70% 3min → aquation 5min.
3. antigen retrieval: hot high pressure reparation
0.01M sodium citrate buffer (PH6.0) is added in pressure cooker.Liquid level is set not have slide tissue on staining rack. The lid of stainless steel pressure cooker is covered, is slowly heated (about 5min), minor valve, which will rise, to be come.After 1~2min, heat source is left, Room temperature to be down to opens lid (avoiding supercooling from overheating, to prevent flake).
4. dyeing: SABC method
3%H2O2It is incubated for 10min in room temperature and (if any decortication, uses 0.3%H2O2)；PBS is rinsed, 5min × 3 time；It is added dropwise 5% BSA confining liquid, room temperature wet box is interior to be incubated for 30min, inclines, does not wash；Primary antibody 40ul is added dropwise: rabbit-anti rat NF-KBp65 monoclonal is anti- Body (1:100), 4 DEG C overnight；PBS is rinsed, 5min × 3 time；Secondary antibody working solution is added dropwise, is incubated for 30min in room temperature wet box；PBS punching It washes, 5min × 3 time；Reagent SABC is added dropwise, is incubated for 30min in room temperature wet box；PBS is rinsed, 5min × 3 time；DAB colour developing: under mirror Grasp colour developing degree；Distilled water flushing；Haematoxylin redyes 5min, and clear water rinses；Hydrochloride alcohol, ammonium hydroxide differentiation；Dehydration: 70% second The ethyl alcohol of alcohol 5min → 80% ethyl alcohol of 5min → 95% 5min → dehydrated alcohol II5min → dehydrated alcohol I5min；It is transparent: dimethylbenzene II3min → dimethylbenzene I3min；Neutral gum mounting；Microscopy.
3. ImmunohistochemistryResults Results determine immunohistochemistry scoring (immunohistochemical scores, HIS) with cell Film, cytoplasm, to occur faint yellow or brown yellow granule in nucleus be positive expression, and staining power is higher than the non-specificity of background Dyeing is determined as positive cell.Every histotomy randomly selects 5 low power (× 100) visuals field, is calculated using secondary weighted method Positive overall score, i.e., positive overall score=staining power scoring × staining positive cells rate scoring.
Staining power Scoring System: not colored is 0 point, and faint yellow is 1 point, and brown or sepia are 2 points, brown or dark brown Color is 3 points.
The scoring of staining positive cells rate: accounting for 0-9% is 0 point, and 10-24% is 1 point, and 25-49% is 2 points, 50-100% 3 Point.
Positive overall score 0 divide for-；1-3 point for+；4-6 points are ++；7-9 points are +++.
4. statistical analysis
Statistical procedures are carried out using SPSS16.0 software.Measurement data is with mean ± standard deviationIt indicates, group Between mean comparison use one-way analysis of variance, P < 0.05 be difference it is statistically significant.Ranked data are analyzed with Ridit, 95% credibility interval of the Ridit value of detection group does not include 0.5, P < 0.05, and difference is statistically significant.Statistical result is as follows:
1) influence that the Chinese herbal granules of embodiment 3 express c reactive protein
Model group c reactive protein obviously increases (P < 0.05) compared with sham-operation group；The Chinese herbal granules of embodiment 3 are respectively treated Group significantly reduces c reactive protein expression (P < 0.05) compared with model group with Tongxinluo treatment group.Be shown in Table 7, Fig. 7, Fig. 7 indicate CRP, The case where expression of IL-6 and TNF-α.
The influence that table 7 expresses c reactive protein
* the P=0.000 compared with sham-operation group；△ P value compared with model group is followed successively by 0.05,0.003,0.004,0.000
2) influence of the Chinese herbal granules of embodiment 3 to IL-6, TNF-α expression
Model group significantly increases (P < 0.05) compared with sham-operation group IL-6, TNF-α concentration, and the Chinese herbal granules of embodiment 3 are each Dosage group significantly reduces IL-6, TNF-α concentration (P < 0.05), the no significant difference compared with Tongxinluo group compared with model group.Be shown in Table 8, Fig. 7.
The influence that table 8 expresses IL-6, TNF-a
* the P=0.000 compared with sham-operation group；△ P value compared with model group is followed successively by 0.000,0.000,0.000, 0.001；# P=0.016 compared with sham-operation group；@P value compared with model group is followed successively by 0.009,0.013,0.002,0.035.
3) Chinese herbal granules of embodiment 3 are to TXA2、PGI2The influence of expression
Compared with sham-operation group, model group PGI2Level is substantially reduced；TXA2Horizontal apparent increase (P < 0.05).Embodiment Each dosage group of 3 Chinese herbal granules dramatically increases PGI compared with model group with Tongxinluo group2It is horizontal；Reduce TXA2Level (P < 0.05).It is shown in Table 9, Fig. 8, Fig. 8 shows TXA2And PGI2The case where expression.
Table 9 is to TXA2、PGI2The influence of expression
* the P=0.000 compared with sham-operation group；△ P value compared with model group is followed successively by 0.034,0.008,0.008, 0.000；# P=0.000 compared with sham-operation group；@P value compared with model group is followed successively by 0.000,0.000,0.000,0.000
4) Chinese herbal granules of embodiment 3 are expressed cardiac muscle cell NF-KBp65 and are influenced
DAB is dyed, and NF-KB immunohistochemistry positive expression situation in rat myocardial cell is observed under microscope (× 100).Knot Fruit shows: the NF-KB positive expression in the model group cardiac muscle cell of embodiment 3 is obviously increased compared with sham-operation group, positive integral warp Ridit analyzes comparing difference significantly (P < 0.05), and Chinese herbal granules low, middle and high dose groups and Tongxinluo group positive expression are obvious Weaken, there were significant differences compared with model group (P < 0.05) for positive integral.Experimental result is shown in Table 3-4, Fig. 9, table 3-4 and Fig. 9 table The case where showing NF-KB immunohistochemistry positive expression in different experiments group rat myocardial cell；Wherein Fig. 9-a is sham-operation group；Figure 9- is model group；Fig. 9-c is low dose group；Fig. 9-d is middle dose group；Fig. 9-e is high dose group；9-f is positive drug group.
Influence of the table 3-4 to NF-KB immunohistochemistry positive expression in rat myocardial cell
* compared with sham-operation group, 95% credibility interval (0.8392,1.0501) of model group Ridit value；Low dose group 95% credibility interval (0.7279,0.9388) of Ridit value；Middle dose group Ridit value 95% credibility interval (0.5962, 0.8071)；95% credibility interval (0.5235,0.7343) of high dose group Ridit value；The 95% of Tongxinluo group Ridit value can Believe section (0.6257,0.8365), above does not include 0.5, P < 0.05.
# is compared with model group, 95% credibility interval (0.2046,0.4154) of low dose group Ridit value；Middle dose group 95% credibility interval (0.2211,0.4319) of Ridit value；High dose group Ridit value 95% credibility interval (0.1471, 0.3580)；95% credibility interval (0.1849,0.3958) of Tongxinluo group Ridit value, above does not include 0.5, P < 0.05.
This experiment by immunohistochemical observation to model group cardiac muscle NF-KB expression significantly and sham-operation group, in embodiment 3 Medicine granule and Tongxinluo group can inhibit NF-KB to express.The Chinese herbal granules of embodiment 3 may be by inhibiting NF-KB thin Extracellular positive feedback activated pathway, to realize protection Myocardial Effects.As shown in the above, the Chinese medicine of the embodiment of the present application 3 Granule can significantly reduce c reactive protein content, to reduce blood vessel endothelium injury and degree of myocardial ischemia；Serum can also be reduced IL-6, TNF-α are horizontal, prompt have inhibition inflammatory reaction effect, may prevent in cardiac muscle to formation, the rupture for preventing patch Chrotoplast damage tool has certain effect；TXA can also be reduced2Concentration increases PGI2Concentration, so as to alleviate vasopasm, reduce The formation of thrombus is played the role of protecting cardiac muscle cell；Moreover it is possible to which cardiac muscle cell NF-KB is inhibited to express, it may be possible to pass through suppression The extracellular positive feedback activated pathway of NF-KB processed, to realize protection Myocardial Effects.
As can be seen from the above embodiments, the electrocardio of the substantially reduced Acute Myocardial Ischemia Rats of Chinese herbal granules energy of embodiment 3 Figure variation degree, reducing myocardial infarction area reduce Serum CK-MB, LDH, AST and cTnI activity, the variation of tectology It is consistent with the change of Myocardial Enzymologic, show that the Chinese herbal granules of embodiment 3 have protective effect to ischemic myocardium.Secondly, real The Chinese herbal granules for applying example 3 can significantly reduce c reactive protein, IL-6, TNF-α content, prompt have inhibition inflammatory reaction effect, Formation, rupture to patch is prevented；Prevent myocardium endothelial cell damage tool from having certain effect.Again, the Chinese medicine of embodiment 3 Granula can reduce TXA2Concentration increases PGI2Concentration shows that it can obviously inhibit the increase of internal vaso-excitor material, keeps PGI2/ TXA2Physiological equilibrium plays increase myocardial blood supply so as to the formation alleviated vasopasm, reduce thrombus, protects cardiac muscle cell Effect.In addition, the Chinese herbal granules of embodiment 3 may reduce the heart by inhibiting the extracellular positive feedback activated pathway of NF-KB Myocyte NF-KB activation, to reduce the genetic transcription of inflammatory factor.In summary, Chinese medicinal granule of the present invention through embodiment 3 Agent has good function of resisting myocardial ischemia, and mechanism of action inhibits modulating vascular endothelial inflammation access related to it.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered It is considered as protection scope of the present invention.

Claims (10)

1. a kind of Chinese medicine composition for treating acute coronary syndrome, the raw material including following parts by weight: 20~50 parts of honeysuckle, 20~50 parts of root of kirilow rhodiola, 5~20 parts of leech, 10~50 parts of Snakegourd Fruit, 10~30 parts of radix scrophulariae, 20~40 parts of Radix Angelicae Sinensis, rhizoma nardostachyos 10~30 Part, 20~40 parts of Radix Salviae Miltiorrhizae and 5~10 parts of Radix Glycyrrhizae.
2. Chinese medicine composition according to claim 1, which is characterized in that the Chinese medicine composition includes following parts by weight Raw material: 30~40 parts of honeysuckle, 30~40 parts of root of kirilow rhodiola, 10~15 parts of leech, 20~40 parts of Snakegourd Fruit, 15~25 parts of radix scrophulariae, when Return 25~35 parts, 15~25 parts of rhizoma nardostachyos, 25~35 parts of Radix Salviae Miltiorrhizae and 6~9 parts of Radix Glycyrrhizae.
3. the preparation method of Chinese medicine composition of any of claims 1 or 2, comprising the following steps:
1) Radix Salviae Miltiorrhizae is mixed with ethanol water, carries out refluxing extraction, filtering collects filtrate concentration, obtains the first thick paste, collects Filter residue is first dregs of a decoction；
2) honeysuckle, Radix Angelicae Sinensis and rhizoma nardostachyos are mixed with water, carries out the first water extraction, collection obtains volatile oil, extracting solution and second The dregs of a decoction；
3) by step 1) first dregs of a decoction, step 2) second dregs of a decoction and root of kirilow rhodiola, Snakegourd Fruit, leech, radix scrophulariae, Radix Glycyrrhizae and water Mixing, carries out the second water extraction, and filtering, concentrated filtrate obtain molten cream；
4) it stands, extracts after mixing step 3) the molten cream with ethanol water, filtering, concentrated filtrate obtain second Thick paste；
5) it is stood after mixing step 1) first thick paste, step 4) second thick paste and the step 2) extracting solution, mistake Filter, dried filtrate, dried object crush to obtain medicinal powder；
6) by volatile oil beta-cyclodextrin inclusion compound described in step 2), inclusion compound is obtained；
7) inclusion compound described in step 6) is mixed with medicinal powder described in step 5), obtains Chinese medicine composition；
There is no temporal restriction relations for step 5) and step 6).
4. preparation method according to claim 3, which is characterized in that the matter of Radix Salviae Miltiorrhizae and ethanol water in the step 1) Amount is than being 1:(7~9).
5. preparation method according to claim 3 or 4, which is characterized in that ethyl alcohol in ethanol water in the step 1) Percent by volume be 80%~95%.
6. preparation method according to claim 3, which is characterized in that the temperature of the refluxing extraction in the step 1) is 60 ~80 DEG C；The number of the refluxing extraction is 2~4 times.
7. preparation method according to claim 3, which is characterized in that honeysuckle in the step 2), Radix Angelicae Sinensis and rhizoma nardostachyos The mass ratio of gross mass and water is 1:(9~11).
8. preparation method according to claim 3, which is characterized in that the temperature that the second water extracts in the step 3) is 80 ~100 DEG C；The number that second water extracts is 2~4 times；The time extracted every time is 1~3h.
9. preparation method according to claim 3, which is characterized in that the temperature stood in the step 5) is 1~5 DEG C； The time of the standing is 18~30h.
10. preparation method according to claim 3, which is characterized in that the partial size of the step 5) traditional Chinese medicine powder is 80~100 Mesh.
CN201910007680.2A 2019-01-04 2019-01-04 A kind of Chinese medicine composition and preparation method thereof for treating acute coronary syndrome Withdrawn CN109568448A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111544465A (en) * 2020-06-08 2020-08-18 上海市同济医院 Traditional Chinese medicine composition for secondary prevention after PCI (peripheral component interconnect) operation of acute coronary syndrome

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111544465A (en) * 2020-06-08 2020-08-18 上海市同济医院 Traditional Chinese medicine composition for secondary prevention after PCI (peripheral component interconnect) operation of acute coronary syndrome
CN111544465B (en) * 2020-06-08 2022-03-25 上海市同济医院 Traditional Chinese medicine composition for secondary prevention after PCI (peripheral component interconnect) operation of acute coronary syndrome

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