Traditional Chinese medicine composition for improving microcirculation and preparation method and application thereof

Abstract

The invention provides a traditional Chinese medicine composition for improving microcirculation as well as a preparation method and application thereof, and relates to the technical field of medicines. The preparation method of the traditional Chinese medicine composition specifically comprises the following steps: S1, alcohol extraction and concentration of rhizoma corydalis to obtain filter residues and a concentrated solution 1; s2, adding alkali liquor to soak the filter residues obtained in the step S1 to obtain alkalized filter residues; mixing alkalized filter residues with chloroform, heating and extracting, and carrying out reduced pressure distillation on an extracting solution to obtain a concentrated solution 2; s3, extracting the szechwan chinaberry fruit with alcohol, and concentrating to obtain filter residues and Washing the filtrate with a solvent, extracting and concentrating to obtain a concentrated solution 3; and S4, adding water into the concentrated solution 1, the concentrated solution 2 and the concentrated solution 3, mixing, performing enzymolysis, and filtering to obtain the composition. The composition prepared by the specific preparation method disclosed by the invention is high in stability, high in bioavailability and better in drug effect, and has a remarkable effect of improving microcirculation.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a traditional Chinese medicine composition for improving microcirculation, its preparation method and application. Background Art

[0002] Microcirculation refers to the blood circulation between arterioles and venules, which is the place for the exchange of substances between blood and tissue cells. The basic function of microcirculation is to carry out the exchange of substances between blood and tissue fluid. Under normal circumstances, the blood flow in microcirculation adapts to the metabolic level of tissues and organs, ensuring the blood perfusion of each tissue and organ and regulating the volume of blood returning to the heart. If microcirculation disorders occur, it will directly affect the physiological functions of each organ. Microcirculation supplies oxygen and nutrients to tissue cells and takes away metabolic wastes, ensuring the normal life activities. Microcirculation disorders are involved in the occurrence of many diseases, such as acute inflammation, trauma, burns, shock, chronic ulcer disease, hepatitis, cirrhosis, senile hypertension, diabetes, cardiovascular and cerebrovascular diseases, etc. Therefore, improving human microcirculation plays an extremely important role in regulating human health.

[0003] Modern medicine generally uses methods such as protecting microvascular structure (methods: anti-oxidative stress, anti-inflammatory), improving hemodynamics (methods: anti-platelet aggregation, reducing microvascular permeability) and blood rheology (methods: anti-platelet aggregation, reducing microvascular permeability, promoting blood circulation to remove blood stasis) to improve microcirculation disorders and make interventions for specific pathological links. However, the above intervention methods all involve Western medicine treatment, and long-term and large-dose use all have side effects to varying degrees.

[0004] There are many studies on traditional Chinese medicine compositions for improving microcirculation in the prior art. "Research Progress on the Experiment of Traditional Chinese Medicine in Improving Microcirculation" (Shi Da, Liu Hong, Yuan Qionghong, Yuan Hao) emphasizes that traditional Chinese medicine has significant effects on improving microcirculation disorders.

[0005] Chinese Patent CN117462647A discloses a composition for improving microcirculation, which is made of the following components in parts by weight: 30 - 100 parts of ginseng, 100 - 150 parts of Ficus hirta Vahl, 10 - 20 parts of cinnamon, 20 - 50 parts of licorice, and 30 - 70 parts of ginger. This composition has food-grade safety. Although it has small side effects, its onset is slow.

[0006] Chinese Patent CN107648422A relates to a traditional Chinese medicine with the efficacy of promoting blood circulation and removing blood stasis, which is made from the following medicinal materials: including peach kernels, salvia miltiorrhiza, angelica sinensis, folium isatidis, radix paeoniae rubra, achyranthes bidentata, platycodon grandiflorum, toosendan fruit, fructus aurantii, liquorice, pseudostellaria heterophylla, and Chinese yam; it has the advantages of promoting blood circulation and removing blood stasis, improving microcirculation, and promoting qi and relieving pain. Chinese Patent CN104189834A discloses a traditional Chinese medicine for dysmenorrhea of yang deficiency and internal cold type. The composition and weight of the traditional Chinese medicine are as follows: 12 grams of ginseng, 12 grams of donkey-hide gelatin, 6 grams of Corydalis yanhusuo, 9 grams of angelica sinensis, 6 grams of chuanxiong rhizome, 6 grams of pinellia ternata, 9 grams of ginger, 9 grams of evodia rutaecarpa, and 12 grams of ophiopogon japonicus. The effects of this invention are: convenient material collection, quick effect, regulating the physiological function of the uterus, improving microcirculation, enhancing immunity, and having the efficacy of warming the meridian and dispelling cold, and warming the uterus and relieving pain. However, the components of the compositions involved in the above patents are 9 - 12 kinds, which are too complicated. Developing a composition with simple components, small side effects, and significantly improving microcirculation is the research focus in this field.

[0007] Toosendan fruit is the dried ripe fruit of Melia toosendan Sieb. et Zucc. of the Meliaceae family. Its nature and flavor are bitter and cold, and it belongs to the liver, small intestine, and bladder meridians, having the efficacy of dispersing the stagnated liver qi, promoting qi and relieving pain, and expelling worms. It is commonly used to treat symptoms such as stagnation of liver qi transforming into fire, chest and hypochondrium fullness and pain, abdominal pain due to hernia, and abdominal pain due to worm accumulation. Corydalis yanhusuo is the dried tuber of Corydalis yanhusuo W. T. Wang of the Papaveraceae family. Its nature and flavor are pungent, bitter, and warm, and it belongs to the liver and spleen meridians, having the efficacy of promoting blood circulation to remove stasis and promoting qi and relieving pain. Corydalis yanhusuo is a commonly used analgesic in traditional Chinese medicine clinical practice, especially having a significant effect on pain caused by qi stagnation and blood stasis. However, at present, there is less in-depth research on the composition with toosendan fruit and Corydalis yanhusuo as raw materials in terms of improving microcirculation.

[0008] Based on the existing pharmacological activities of toosendan fruit and Corydalis yanhusuo, by deeply studying their extraction methods and developing their new clinical efficacy, obtaining a composition with simple components, low side effects, and good efficacy is the research focus of researchers in this field. Summary of the Invention

[0009] In view of the above problems, the present invention provides a traditional Chinese medicine composition for improving microcirculation and its preparation method. Through a large number of studies, the results show that the composition of toosendan fruit and Corydalis yanhusuo under the specific extraction method of the present invention has a good curative effect on the treatment of improving microcirculation.

[0010] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0011] On the one hand, the present invention provides a preparation method of a traditional Chinese medicine composition for improving microcirculation, comprising the following steps:

[0012] S1: Extract Corydalis yanhusuo with alcohol, concentrate, to obtain filter residue and concentrated solution 1;

[0013] S2: The filter residue obtained in S1 is soaked in an alkaline solution to obtain an alkalized filter residue; the alkalized filter residue is mixed with chloroform, heated for extraction, and the extract is distilled under reduced pressure to obtain concentrated solution 2.

[0014] S3: The Szechwan Chinaberry Fruit is extracted with alcohol, concentrated to obtain a filter residue and a filtrate; the filtrate is washed with a solvent, extracted, and concentrated to obtain concentrated solution 3.

[0015] S4: Concentrated solution 1, concentrated solution 2, and concentrated solution 3 are mixed with water, enzymatically hydrolyzed, and filtered to obtain a traditional Chinese medicine composition.

[0016] Preferably, in S1, the alcohol for alcohol extraction is ethanol or methanol; more preferably, in S1, the alcohol for alcohol extraction is ethanol.

[0017] More preferably, in S1, the volume fraction of the alcohol for alcohol extraction is 60%-90%. Even more preferably, in S1, the volume fraction of the alcohol is 80%.

[0018] Preferably, in S1, the material-liquid ratio of Corydalis Rhizoma to alcohol is 1 g: 10-12 L.

[0019] Preferably, in S1, the number of times of alcohol extraction is 1-3 times; more preferably, in S1, the number of times of alcohol extraction is 3 times.

[0020] Preferably, in S1, the temperature of alcohol extraction is 70-90 °C, and the time is 1-3 h; more preferably, in S1, the temperature of alcohol extraction is 80 °C, and the time is 3 h.

[0021] Preferably, in S2, the alkaline solution is a mixture of 8-12 wt% ammonia water and 1-5 wt% sodium carbonate solution.

[0022] More preferably, in S2, the alkaline solution is a mixture of 8-10 wt% ammonia water and 3-5 wt% sodium carbonate solution.

[0023] Even more preferably, in S2, the alkaline solution is a mixture of 10 wt% ammonia water and 5 wt% sodium carbonate solution.

[0024] Preferably, in S2, the material-liquid ratio of the filter residue to the alkaline solution is 1 g: 8-12 L; more preferably, in S2, the material-liquid ratio of the filter residue to the alkaline solution is 1 g: 8-10 L. Even more preferably, in S2, the material-liquid ratio of the filter residue to the alkaline solution is 1 g: 10 L.

[0025] Preferably, in S2, the material-liquid ratio of the alkalized filter residue to chloroform is 1 g: 10-15 L; more preferably, in S2, the material-liquid ratio of the alkalized filter residue to chloroform is 1 g: 10-12 L; even more preferably, in S2, the material-liquid ratio of the alkalized filter residue to chloroform is 1 g: 12 L.

[0026] Preferably, in S2, the temperature for heating extraction is 80 - 90°C, and the time is 1 - 3 h; more preferably, in S2, the temperature for heating extraction is 85 - 90°C, and the time is 1 - 2 h; most preferably, in S2, the temperature for heating extraction is 90°C, and the time is 1.5 h.

[0027] Preferably, in S2, the number of times of heating extraction is 1 - 3 times; more preferably, in S2, the number of times of heating extraction is 3 times.

[0028] Preferably, in S3, the alcohol for alcohol extraction is ethanol.

[0029] More preferably, the ethanol has a volume fraction of 45% - 65%; most preferably, the ethanol has a volume fraction of 60%.

[0030] Preferably, in S3, the temperature for alcohol extraction is 70 - 90°C, and the time is 1 - 2 h; more preferably, in S3, the temperature for alcohol extraction is 80°C, and the time is 1.5 h.

[0031] Preferably, in S3, the material - liquid ratio of Szechwan Chinaberry Fruit to alcohol is 1 g: 10 - 12 L.

[0032] Preferably, in S3, the solvent is selected from petroleum ether or n - hexane; more preferably, in S3, the solvent is petroleum ether.

[0033] Preferably, in S3, the solvent for extraction is ethyl acetate.

[0034] Preferably, in S4, the enzyme for enzymatic hydrolysis is selected from at least one of cellulase, esterase, and β - glucosidase.

[0035] More preferably, in S4, the enzyme for enzymatic hydrolysis is a mixture of cellulase, esterase, and β - glucosidase.

[0036] Preferably, the mass ratio of cellulase, esterase, and β - glucosidase is 1: 0.5 - 0.8: 0.2 - 0.4.

[0037] More preferably, the mass ratio of cellulase, esterase, and β - glucosidase is 1: 0.6: 0.2.

[0038] Preferably, the enzyme activity of the cellulase is

[0039] Preferably, in S4, the addition amount of water is 3 - 5 times the total mass of concentrated solution 1, concentrated solution 2, and concentrated solution 3; more preferably, in S4, the addition amount of water is 5 times the total mass of concentrated solution 1, concentrated solution 2, and concentrated solution 3.

[0040] Preferably, in S4, the mass of the enzyme for enzymatic hydrolysis is 0.1-1% of the total mass of concentrated solution 1, concentrated solution 2 and concentrated solution 3; More preferably, in S4, the mass of the enzyme for enzymatic hydrolysis is 0.5% of the total mass of concentrated solution 1, concentrated solution 2 and concentrated solution 3.

[0041] Preferably, in S4, the pH of the enzymatic hydrolysis is 4.5-5, the temperature is 40-45°C, and the time is 1-3 h.

[0042] More preferably, in S4, the pH of the enzymatic hydrolysis is 5, the temperature is 45°C, and the time is 2 h.

[0043] On the other hand, the present invention provides a traditional Chinese medicine composition prepared by the above-mentioned preparation method.

[0044] On yet another aspect, the present invention provides the use of the traditional Chinese medicine composition prepared by the above-mentioned preparation method in the preparation of a drug for improving microcirculation.

[0045] Preferably, the drug can be prepared into dosage forms for administration such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, troches, etc., and suitable drug carriers in the art can be selected for different dosage forms.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] 1. By further alkalizing and extracting the residue of Corydalis yanhusuo after ethanol extraction, the present invention extracts the alkaloid components therein, such as tetrahydropalmatine, corydaline, protopine and corypalmine, to the greatest extent, and significantly improves its effect of improving blood circulation.

[0048] 2. By washing the extract of Toosendan fruit after ethanol extraction with petroleum ether, the present invention can wash away toxic substances such as toosendanin therein.

[0049] 3. Through the specific enzymatic hydrolysis of the present invention, under the synergistic effect of cellulase, esterase and β-glucosidase, the obtained composition has a significant effect of improving microcirculation.

[0050] 4. In summary, the composition prepared by the specific preparation method of the present invention has high stability, high bioavailability, better drug efficacy, and a significant effect of improving microcirculation. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 It is the weight diagram of rats in the blank group, model group, example 1 group and positive control group during the administration period.

[0052] Figure 2 It is the detection result diagram of blood rheology of rats in the blank group, model group, example 1 group and positive control group, whereinFigure 2 A is the whole blood viscosity graph at low shear (10 s -1 ); Figure 2 B is the whole blood viscosity graph at medium shear (50 s -1 ); Figure 2 C is the whole blood viscosity graph at high shear (200 s -1 ); Figure 2 D is the plasma viscosity graph. Compared with the blank group, * P < 0.05, ** P < 0.01; compared with the model group, # P < 0.05, ## P < 0.01.

[0053] Figure 3 are the graphs of the four coagulation tests of rats in the blank group, model group, Example 1 group, and positive control group. Among them, Figure 3 A is the fibrinogen (FIB) graph; Figure 3 B is the thrombin time (TT) graph; Figure 3 C is the prothrombin time (PT) graph; Figure 3 D is the activated partial thromboplastin time (APTT); compared with the blank group, * P < 0.05, ** P < 0.01; compared with the model group, # P < 0.05, ## P < 0.01. Specific implementation manners

[0054] In order to make the technical means, creative features, achieved purposes, and functions of the present invention easy to understand, the present invention will be further clarified below in conjunction with specific embodiments. However, the following embodiments are only the preferred embodiments of the present invention and not all of them. Based on the embodiments in the implementation manners, other embodiments obtained by those skilled in the art without creative efforts all belong to the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all ordinary commercially available products, and no specific limitation is made on their sources. The technical and scientific terms used in the embodiments have the meanings commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0055] Raw materials:

[0056] Toosendan Fruit (batch number: C1073111, Traditional Chinese Medicine Decoction Pieces Factory of Guangdong Provincial Medicinal Materials Company);

[0057] Rhizoma Corydalis (batch number: Y1323513, Traditional Chinese Medicine Decoction Pieces Factory of Guangdong Provincial Medicinal Materials Company);

[0058] Adrenaline Hydrochloride Injection (batch number: 240312, Shanghai Quanyu Biotech Animal Pharmaceutical Co., Ltd.);

[0059] 0.9% Sodium Chloride Injection (Batch No.: 1J73B3, China Otsuka Pharmaceutical Co., Ltd.);

[0060] Thrombin Time (TT) Reagent (Batch No.: Y20220801, Chengdu Aixuelun Medical Technology Co., Ltd.);

[0061] Prothrombin Time (PT) Reagent (Batch No.: Y20230401, Chengdu Aixuelun Medical Technology Co., Ltd.);

[0062] Fibrinogen (FIB) Reagent (Batch No.: Y20230530, Chengdu Aixuelun Medical Technology Co., Ltd.);

[0063] Activated Partial Thromboplastin Time (APTT) Reagent (Batch No.: Y20230401, Chengdu Aixuelun Medical Technology Co., Ltd.);

[0064] Calcium Chloride Reagent (CaCl) (Batch No.: Y20230401, Chengdu Aixuelun Medical Technology Co., Ltd.);

[0065] Cellulase (Catalog No.: S10041, Shanghai Yuanye Bio-Technology Co., Ltd.)

[0066] Esterase (Catalog No.: S23705, Shanghai Yuanye Bio-Technology Co., Ltd.)

[0067] β-Glucosidase (Catalog No.: S10047, Shanghai Yuanye Bio-Technology Co., Ltd.)

[0068] Instrument:

[0069] Automatic Blood Rheology Dynamic Analyzer (Model: MEN-C100A, Jinan Meiyilin Electronic Instrument Co., Ltd.);

[0070] Automatic Coagulation Analyzer (Model: EC6800, Chengdu Aixuelun Medical Technology Co., Ltd.).

[0071] Example 1

[0072] A preparation method of a traditional Chinese medicine composition for improving microcirculation, specifically:

[0073] The mass ratio of Corydalis Rhizoma to Toosendan Fruit is 1:1;

[0074] S1: Mix Corydalis Rhizoma with an ethanol solution with a volume fraction of 80%, the material-liquid ratio is 1 g: 10 L, extract with alcohol at 80 °C for 3 h, repeat 3 times, combine the alcohol extracts, concentrate, to obtain filter residue and concentrated solution 1;

[0075] S2: The filter residue obtained from S1 is soaked in an alkaline solution for 20 - 40 min. The alkaline solution is a mixture of 10 wt% ammonia water and 5 wt% sodium carbonate solution. The material - liquid ratio of the filter residue to the alkaline solution is 1 g:10 L to obtain an alkalized filter residue;

[0076] The alkalized filter residue is mixed with chloroform. The material - liquid ratio of the alkalized filter residue to chloroform is 1 g:12 L. It is heated to 90 °C for extraction for 1.5 h, and the extraction is repeated 3 times. The extraction liquids are combined and distilled under reduced pressure to obtain concentrated liquid 2;

[0077] S3: Toosendan fruit is mixed with an ethanol solution with a volume fraction of 60%. The material - liquid ratio is 1 g:10 L. It is extracted with ethanol at 80 °C for 1 h, and the extraction is repeated 3 times. The ethanol - extracted liquids are combined and concentrated to obtain a filter residue and a filtrate;

[0078] The filtrate is washed 3 times with petroleum ether, extracted with ethyl acetate, and concentrated to obtain concentrated liquid 3;

[0079] S4: Concentrated liquid 1, concentrated liquid 2, and concentrated liquid 3 are combined to obtain a mixture. 5 times the mass of water is added to the mixture and mixed. An enzyme (a mixture of cellulase, esterase, and β - glucosidase, and the mass ratio of the three is 1:0.6:0.2) accounting for 0.5% of the mass of the mixture is added for enzymatic hydrolysis. The pH of the enzymatic hydrolysis is 5, the temperature is 45 °C, and the time is 2 h. After filtration and drying, a traditional Chinese medicine composition is obtained.

[0080] Example 2

[0081] A preparation method of a traditional Chinese medicine composition for improving microcirculation is as follows:

[0082] The mass ratio of Corydalis yanhusuo to Toosendan fruit is 1:1;

[0083] S1: Corydalis yanhusuo is mixed with an ethanol solution with a volume fraction of 60%. The material - liquid ratio is 1 g:12 L. It is extracted with ethanol at 90 °C for 3 h, and the extraction is repeated 3 times. The ethanol - extracted liquids are combined and concentrated to obtain a filter residue and concentrated liquid 1;

[0084] S2: The filter residue obtained from S1 is soaked in an alkaline solution for 20 - 40 min. The alkaline solution is a mixture of 8 wt% ammonia water and 1 wt% sodium carbonate solution. The material - liquid ratio of the filter residue to the alkaline solution is 1 g:8 L to obtain an alkalized filter residue;

[0085] The alkalized filter residue is mixed with chloroform. The material - liquid ratio of the alkalized filter residue to chloroform is 1 g:15 L. It is heated to 80 °C for extraction for 1 h, and the extraction is repeated 3 times. The extraction liquids are combined and distilled under reduced pressure to obtain concentrated liquid 2;

[0086] S3: Toosendan fruit is mixed with an ethanol solution with a volume fraction of 45%. The material - liquid ratio is 1 g:12 L. It is extracted with ethanol at 70 °C for 2 h, and the extraction is repeated 3 times. The ethanol - extracted liquids are combined and concentrated to obtain a filter residue and a filtrate;

[0087] The filtrate was washed 3 times with petroleum ether, extracted with ethyl acetate, and concentrated to obtain concentrated solution 3;

[0088] S4: Concentrated solutions 1, 2, and 3 were combined to obtain a mixture, which was mixed with 3 times the mass of water of the mixture, and an enzyme (a mixture of cellulase, esterase, and β-glucosidase, with a mass ratio of 1:0.5:0.4 for the three) accounting for 0.1% of the mass of the mixture was added for enzymatic hydrolysis. The pH of the enzymatic hydrolysis was 4.5, the temperature was 40 °C, and the time was 3 h. After filtration and drying, a traditional Chinese medicine composition was obtained.

[0089] Example 3

[0090] A preparation method of a traditional Chinese medicine composition for improving microcirculation is specifically as follows:

[0091] The mass ratio of Corydalis Rhizoma to Toosendan Fruit is 1:1;

[0092] S1: Corydalis Rhizoma was mixed with an ethanol solution with a volume fraction of 90%, the material-liquid ratio was 1 g:8 L, and ethanol extraction was carried out at 70 °C for 3 h. This was repeated 3 times, and the ethanol extracts were combined and concentrated to obtain a filter residue and concentrated solution 1;

[0093] S2: The filter residue obtained in S1 was soaked in an alkali solution for 20 - 40 min. The alkali solution was a mixture of 12 wt% ammonia water and 3 wt% sodium carbonate solution, and the material-liquid ratio of the filter residue to the alkali solution was 1 g:12 L to obtain an alkalized filter residue;

[0094] The alkalized filter residue was mixed with chloroform, and the material-liquid ratio of the alkalized filter residue to chloroform was 1 g:10 L. It was heated to 90 °C for extraction for 3 h. This was repeated 3 times, and the extraction liquids were combined and distilled under reduced pressure to obtain concentrated solution 2;

[0095] S3: Toosendan Fruit was mixed with an ethanol solution with a volume fraction of 65%, the material-liquid ratio was 1 g:10 L, and ethanol extraction was carried out at 85 °C for 1.5 h. This was repeated 3 times, and the ethanol extracts were combined and concentrated to obtain a filter residue and a filtrate;

[0096] The filtrate was washed 3 times with petroleum ether, extracted with ethyl acetate, and concentrated to obtain concentrated solution 3;

[0097] S4: Concentrated solutions 1, 2, and 3 were combined to obtain a mixture, which was mixed with 5 times the mass of water of the mixture, and an enzyme (a mixture of cellulase, esterase, and β-glucosidase, with a mass ratio of 1:0.8:0.3 for the three) accounting for 1% of the mass of the mixture was added for enzymatic hydrolysis. The pH of the enzymatic hydrolysis was 5, the temperature was 45 °C, and the time was 2 h. After filtration and drying, a traditional Chinese medicine composition was obtained.

[0098] Comparative Example 1

[0099] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the difference is only that step S2 is removed, specifically as follows:

[0100] The mass ratio of Corydalis Rhizoma to Toosendan Fruit is 1:1;

[0101] S1: Mix Corydalis Rhizoma with an ethanol solution having a volume fraction of 80%, with a material-liquid ratio of 1 g:10 L, perform ethanol extraction at 80 °C for 3 h, repeat 3 times, combine the ethanol extracts, concentrate, to obtain filter residue and concentrated solution 1;

[0102] S2: Mix Toosendan Fruit with an ethanol solution having a volume fraction of 60%, with a material-liquid ratio of 1 g:10 L, perform ethanol extraction at 80 °C for 1 h, repeat 3 times, combine the ethanol extracts, concentrate, to obtain filter residue and filtrate;

[0103] Wash the filtrate with petroleum ether 3 times, add ethyl acetate for extraction, concentrate, to obtain concentrated solution 3;

[0104] S3: Combine concentrated solution 1 and concentrated solution 3 to obtain a mixture, add 5 times the mass of water of the mixture and mix, add an enzyme (a mixture of cellulase, esterase and β-glucosidase, with a mass ratio of the three of 1:0.6:0.2) accounting for 0.5% of the mass of the mixture for enzymatic hydrolysis, the pH of the enzymatic hydrolysis is 5, the temperature is 45 °C, the time is 2 h, filter, dry, to obtain the traditional Chinese medicine composition.

[0105] Comparative Example 2

[0106] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the difference is only in step S3, specifically:

[0107] The mass ratio of Corydalis Rhizoma to Toosendan Fruit is 1:1;

[0108] S1: Mix Corydalis Rhizoma with an ethanol solution having a volume fraction of 80%, with a material-liquid ratio of 1 g:10 L, perform ethanol extraction at 80 °C for 3 h, repeat 3 times, combine the ethanol extracts, concentrate, to obtain filter residue and concentrated solution 1;

[0109] S2: Soak the filter residue obtained in S1 in an alkali solution for 20 - 40 min, the alkali solution is a mixture of 10 wt% ammonia water and 5 wt% sodium carbonate solution, the material-liquid ratio of the filter residue to the alkali solution is 1 g:10 L, to obtain alkalized filter residue;

[0110] Mix the alkalized filter residue with chloroform, the material-liquid ratio of the alkalized filter residue to chloroform is 1 g:12 L, heat to 90 °C for extraction for 1.5 h, repeat extraction 3 times, combine the extraction liquids, perform vacuum distillation, to obtain concentrated solution 2;

[0111] S3: Mix Toosendan Fruit with an ethanol solution having a volume fraction of 60%, with a material-liquid ratio of 1 g:10 L, perform ethanol extraction at 80 °C for 1 h, repeat 3 times, combine the ethanol extracts, concentrate, to obtain filter residue and concentrated solution 3;

[0112] S4: The concentrated liquid 1, concentrated liquid 2 and concentrated liquid 3 are combined to obtain a mixture, 5 times the mass of water is added to the mixture for mixing, and an enzyme (a mixture of cellulase, esterase and β-glucosidase, with the mass ratio of the three being 1:0.6:0.2) accounting for 0.5% of the mass of the mixture is added for enzymatic hydrolysis. The pH of the enzymatic hydrolysis is 5, the temperature is 45°C, the time is 2 h, and then it is filtered and dried to obtain the traditional Chinese medicine composition.

[0113] Comparative Example 3

[0114] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the only difference is that the mass ratio of cellulase, esterase and β-glucosidase in S4 is changed to 0.5:1:0.8.

[0115] Comparative Example 4

[0116] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the only difference is that the enzyme in S4 is changed to cellulase.

[0117] Comparative Example 5

[0118] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the only difference is that the enzyme in S4 is changed to esterase.

[0119] Comparative Example 6

[0120] A preparation method of a traditional Chinese medicine composition for improving microcirculation, compared with Example 1, the only difference is that the enzyme in S4 is changed to β-glucosidase.

[0121] Comparative Example 7

[0122] A preparation method of a traditional Chinese medicine composition for improving microcirculation is as follows:

[0123] Corydalis yanhusuo and Toosendan fruit are mixed in a mass ratio of 1:1, an ethanol solution with a volume fraction of 80% is added, the solid-liquid ratio is 1 g:10 L, and it is extracted with ethanol at 80°C for 3 h. This is repeated 3 times, and the ethanol extracts are combined, concentrated and dried to obtain the traditional Chinese medicine composition.

[0124] Comparative Example 8

[0125] A preparation method of a traditional Chinese medicine composition for improving microcirculation is as follows:

[0126] Corydalis yanhusuo and Toosendan fruit are mixed in a mass ratio of 1:1, decocted with water 2 times. For the first time, 10 times the amount of water is added and decocted for 2 h, and for the second time, 8 times the amount of water is added and decocted for 2 h. The decoction liquids are combined, concentrated and dried to obtain the traditional Chinese medicine composition.

[0127] Test Example

[0128] 1) Animal grouping and administration

[0129] SD rats, 36 - 56 days old, with a body weight of 250 ± 20 g, were fed freely. After 3 days of adaptive feeding, they were randomly divided into a blank group, a model group, a drug - administration group (including Example 1 - 3 groups and Comparative Example 1 - 8 groups), and a positive control group (acetylsalicylic acid group), with 10 rats in each group. The positive control group was intragastrically administered acetylsalicylic acid at a dose of 0.1 g / kg / d; the drug - administration group was intragastrically administered the traditional Chinese medicine composition prepared in Example 1 - 3 and Comparative Example 1 - 8 at a dose of 0.63 g / kg / d, and the blank group and the model group were intragastrically administered an equal amount of distilled water.

[0130] 2) Construction of qi - stagnation and blood - stasis model

[0131] At 1.5 h after the last administration, the qi - stagnation and blood - stasis model was constructed for the rats in the model group and the drug - administration group. The SD rats in the model group and the drug - administration group were subcutaneously injected with 0.1% epinephrine hydrochloride at a dose of 0.8 mg / kg twice in the head and neck, with an interval of 4 hours between the two injections. The blank group was subcutaneously injected with an equal amount of 0.9% sodium chloride injection. After fasting for 12 h without water restriction, all rats were anesthetized by inhaling isoflurane, and blood was taken from the abdominal aorta for the determination of hemorheology and four - item coagulation tests.

[0132] 3) Index detection and model evaluation

[0133] ① Hemorheology detection

[0134] The whole blood viscosity (WBV) of the blood in the heparin - lithium - containing blood collection tube was measured using a fully automatic hemorheometer at low shear rate (10 s -1 ), medium shear rate (50 s -1 ), and high shear rate (200 s -1 ). After the measurement of the whole blood viscosity, the remaining whole blood was centrifuged at 3500 rpm for 10 min and placed on a fully automatic hemorheometer to measure the plasma viscosity (PV).

[0135] ② Four - item coagulation test

[0136] The blood in the sodium citrate blood collection tube was centrifuged at 3500 rpm for 10 min, and the test tube was placed in the sample rack of the EC6800 fully automatic coagulation analyzer. The PT, APTT, FIB, and TT options were selected for the four - item coagulation test.

[0137] ③ Statistical analysis

[0138] IBM SPSS Statistics 26.0 was used to process the experimental data. After excluding the discrepant data within the range of plus and minus two standard deviations, the statistics were performed. With the mean ± standard deviation It was shown that one-way ANOVA was used for comparison among multiple groups, and the results were expressed as P < 0.05 indicating statistically significant differences and P < 0.01 indicating highly statistically significant differences.

[0139] 4) Results

[0140] (1) Characterization and observation

[0141] During the drug administration period, there were no significant differences in the body weights of rats in each group. The body weight table of rats is as follows Figure 1 . Before modeling, the rats in each group were in good condition. After modeling, there were obvious differences in the states of rats in each group. The rats in the blank group were lively and active, with sensitive reactions, white and shiny hair; the rats in the model group were listless, curled up and immobile, with reduced freedom of movement, slower movement, and unkempt and dull hair; the states of the rats in the groups of Examples 1 - 3 were slightly better than those of the rats in the groups of Comparative Examples 1 - 8 and the positive control group, but the states of the rats in the drug administration groups (including the groups of Examples 1 - 3 and Comparative Examples 1 - 8) and the positive control group (acetylsalicylic acid group) were slightly alleviated compared with the symptoms of the model group.

[0142] (2) Hemorheology

[0143] Due to animal deaths during and after the modeling process, the n values were not equal. As shown in Table 1 - 2 and Figure 2 , compared with the blank group, the whole blood viscosity of the model group increased at each shear rate (P < 0.01), showing significant differences, while there were no obvious differences in plasma viscosity changes, indicating that the qi stagnation and blood stasis model was successfully constructed. Compared with the model group, the whole blood viscosity of the traditional Chinese medicine compositions prepared in Examples 1 - 3 and the positive drug group decreased (P < 0.05), showing significant differences, while there were no obvious differences in plasma viscosity changes, indicating that the traditional Chinese medicine compositions prepared in the examples of the present invention have the effect of improving qi stagnation and blood stasis. Compared with the model group, there were no significant differences in the whole blood viscosity of the traditional Chinese medicine compositions prepared in Comparative Examples 1 - 8.

[0144] Table 1. Effects on the whole blood viscosity of rats with qi stagnation and blood stasis

[0145]

[0146] Note: Compared with the blank group, * P < 0.05, ** P < 0.01; compared with the model group, # P < 0.05, ## P < 0.01.

[0147] Table 2. Effects on the plasma viscosity of rats with qi stagnation and blood stasis

[0148]

[0149] (3) Four coagulation tests

[0150] Since animal deaths occurred during and after the model establishment process, some samples could not be detected due to coagulation, so the n values were not equal. As Figure 3 shown in Table 3-6, compared with the blank group, the fibrinogen (FIB) content in the model group was significantly increased (P < 0.05), the activated partial thromboplastin time (APTT) was significantly decreased (P < 0.01), while the prothrombin time (PT) showed a decreasing trend, and the thrombin time (TT) had no obvious change, indicating that the model was successfully constructed; compared with the model group, the fibrinogen content in the groups of Examples 1-3 was significantly decreased (P < 0.05), and the activated partial thromboplastin time was significantly up-regulated (P < 0.01). In the groups of Comparative Examples 1-8, there was a trend of decreasing fibrinogen and a significant up-regulation of the activated partial thromboplastin time (P < 0.05); in the positive drug group, there was a trend of decreasing fibrinogen and a significant up-regulation of the activated partial thromboplastin time (P < 0.01), indicating that the traditional Chinese medicine composition prepared in the examples of the present invention has the effect of improving qi stagnation and blood stasis.

[0151] Table 3. Effects of the traditional Chinese medicine composition on fibrinogen in the four coagulation tests of rats with qi stagnation and blood stasis

[0152]

[0153]

[0154] Note: Compared with the blank group, * P < 0.05, ** P < 0.01; compared with the model group, # P < 0.05, ## P < 0.01.

[0155] Table 4. Effects of the traditional Chinese medicine composition on thrombin time in the four coagulation tests of rats with qi stagnation and blood stasis

[0156] Group n Thrombin time (s) Blank group 9 39.27±1.07 Model group 8 38.69±0.57 Example 1 8 38.93±1.09 Example 2 9 38.54±0.72 Example 3 8 38.72±0.19 Comparative example 1 8 38.64±0.55 Comparative example 2 8 38.90±0.64 Comparative example 3 8 38.85±0.82 Comparative example 4 8 38.70±0.72 Comparative example 5 8 38.73±0.39 Comparative example 6 8 38.69±0.44 Comparative example 7 8 38.70±0.56 Comparative example 8 8 38.54±0.28 Positive control group 9 38.75±0.94

[0157] Note: Compared with the blank group, * P < 0.05, ** P < 0.01; compared with the model group, # P < 0.05, ## P < 0.01.

[0158] Table 5. Effects of the traditional Chinese medicine composition on prothrombin time in the four coagulation tests of rats with qi stagnation and blood stasis

[0159] Group n Prothrombin time (s) Blank group 9 9.88±0.32 Model group 8 9.31±0.36 Example 1 8 9.25±0.81 Example 2 9 9.28±0.57 Example 3 8 9.30±0.65 Comparative example 1 8 9.15±0.28 Comparative example 2 8 9.24±0.26 Comparative example 3 8 9.31±0.63 Comparative example 4 8 9.28±0.54 Comparative example 5 8 9.27±0.33 Comparative example 6 8 9.25±0.42 Comparative example 7 8 9.30±0.50 Comparative example 8 8 9.31±0.63 Positive control group 9 <![CDATA[10.22±0.76 # >

[0160] Note: Compared with the blank group,* P < 0.05, ** P < 0.01; Compared with the model group, # P < 0.05, ## P < 0.01.

[0161] Table 6. Effect on activated partial thromboplastin time in the four coagulation tests of rats with qi stagnation and blood stasis

[0162]

[0163]

[0164] Note: Compared with the blank group, * P < 0.05, ** P < 0.01; Compared with the model group, # P < 0.05, ## P < 0.01.

[0165] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than a limitation on the protection scope of the present invention. Any simple modification or equivalent replacement made by those of ordinary skill in the art to the technical solution of the present invention shall not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a Chinese medicine composition for improving microcirculation, characterized in that: The following steps are involved: S1: extracting Corydalis yanhusuo with alcohol and concentrating it to obtain a filter residue and a concentrated solution 1; S2: The filter residue obtained in S1 is soaked in alkali solution to obtain an alkalized filter residue; the alkalized filter residue is mixed with chloroform, heated for extraction, and the extract is distilled under reduced pressure to obtain a concentrated solution 2; S3: extracting Toosendan fruit with alcohol, concentrating, obtaining filter residue and filtrate; washing the filtrate with solvent, extracting, concentrating, obtaining concentrated solution 3; S4: adding water to the concentrate 1, the concentrate 2 and the concentrate 3, mixing, enzymolyzing, filtering, and obtaining a Chinese medicine composition.

2. The preparation method according to claim 1, characterized in that: In S1, the alcohol for the alcohol extraction is ethanol or methanol; the volume fraction of the alcohol for the alcohol extraction is 60%-90%; the number of alcohol extractions is 1-3 times; the temperature of the alcohol extraction is 70-90° C., and the time is 1-3 hours.

3. The preparation method according to claim 1, characterized in that: In S2, the alkali solution is a mixture of 8-12wt% ammonia water and 1-5wt% sodium carbonate solution; the solid-liquid ratio of the filter residue to the alkali solution is 1g:8-12L; the solid-liquid ratio of the alkalized filter residue to chloroform is 1g:10-15L; the temperature of the heating extraction is 80-90°C, the time is 1-3h; the number of heating extractions is 1-3 times.

4. The preparation method according to claim 1, characterized in that: In S3, the alcohol for the alcohol extraction is an ethanol solution with a volume fraction of 45%-65%; the temperature of the alcohol extraction is 70-90°C, and the time is 1-2h; the solvent is selected from petroleum ether or n-hexane; and the extraction solvent is ethyl acetate.

5. The preparation method according to claim 1, characterized in that: In S4, the enzyme for enzymatic hydrolysis is selected from at least one of cellulase, esterase and β-glucosidase.

6. The preparation method according to claim 5, characterized in that: In S4, the enzyme for enzymolysis is selected from a mixture of cellulase, esterase and β-glucosidase; the mass ratio of the cellulase, esterase and β-glucosidase is 1:0.5-0.8:0.2-0.

4.

7. The preparation method according to claim 1, characterized in that: In S4, the amount of water added is 3-5 times the total mass of the concentrate 1, the concentrate 2 and the concentrate 3; the mass of the enzyme for enzymolysis is 0.1-1% of the total mass of the concentrate 1, the concentrate 2 and the concentrate 3.

8. The preparation method according to claim 1, characterized in that: In S4, the pH of the enzymatic hydrolysis is 4.5-5, the temperature is 40-45°C, and the time is 1-3h.

9. A Chinese medicine composition, characterized in that: It is prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 1 to 8 in preparing a drug for improving microcirculation.

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