A Chinese medicine composition for preventing and treating exercise-induced myocardial injury, and its preparation method and application

Abstract

The present invention provides a Chinese medicine composition for preventing and treating exercise-induced myocardial injury, and a preparation method and application thereof, which belong to the field of medical technology. The Chinese medicine composition for preventing and treating exercise-induced myocardial injury of the present invention is prepared from the following raw materials in parts by weight: 0.8-1.2 parts of total flavonoids of kudzu root, 10-15 parts of kiwi fruit, and 15-20 parts of hawthorn. The Chinese medicine composition of the present invention can reduce the content of MDA in the body during exercise, increase the activity of SOD, thereby increasing the antioxidant capacity of myocardial tissue, reducing the level of lipid peroxidation, and by downregulating the NF-κB signaling pathway, increasing the antioxidant enzyme activity and anti-inflammatory ability of myocardial tissue, and reducing myocardial damage caused by exercise exhaustion.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a Chinese medicine composition for preventing and treating exercise-induced myocardial injury, and a preparation method and application thereof. Background Art

[0002] Exercise-induced myocardial injury refers to myocardial damage caused by high-intensity exercise. Studies have shown that long-distance, high-intensity exercise can cause catecholamine levels in the blood to far exceed normal values, greatly increase myocardial oxygen demand and last for a long time, and ischemia and hypoxia are prone to occur. The mechanism of exercise-induced myocardial injury may be related to the imbalance of oxygen free radicals and myocardial cell apoptosis. During high-intensity exercise, endogenous oxygen free radicals are produced excessively, and the body's clearance of oxygen free radicals is insufficient, resulting in an increase in the content of oxygen free radicals in myocardial cells. Excessive production of free radicals is not only an important apoptosis inducer in the body, but also can trigger lipid peroxidation, enhance lipid peroxidation, and cause an imbalance between oxidative attack and antioxidant capacity, resulting in oxidative stress damage, which is the main cause of exercise-induced myocardial injury. In addition, inflammatory factors produced by inflammatory reactions can directly damage myocardial cells, causing cell structure and dysfunction, and in severe cases directly causing myocardial cell apoptosis and cell necrosis.

[0003] There is no effective means of prevention and treatment of exercise-induced myocardial injury at this stage. Through the investigation of literature, this study believes that traditional Chinese medicine plays a huge advantage in the prevention of exercise-induced myocardial injury before it occurs. In modern Chinese clinical practice, Pueraria root has pharmacological activities of antipyretic and anti-inflammatory, contraction of smooth muscle, dilation of coronary artery vessels to reduce peripheral vascular resistance, increase of cerebral and coronary blood flow, and anti-arrhythmic. It also has the activities of enhancing the body's immunity, lowering blood pressure, lowering blood sugar, anti-oxidation and anti-tumor, so it has a wide range of application value. The chemical components of Pueraria root mainly include flavonoids, coumarins, triterpenoid saponins, alkaloids and other types. Pueraria root total flavonoids contain a large number of isoflavone compounds, which are also the effective part of pharmacological activity. At present, there are few reports on the intervention of Pueraria root total flavonoids in the changes of myocardial oxidative stress and inflammatory response in mice with exhaustive exercise. Summary of the invention

[0004] The purpose of the present invention is to provide a Chinese medicine composition for preventing and treating exercise-induced myocardial injury, and a preparation method and application thereof. The Chinese medicine composition of the present invention can reduce the content of MDA in the body during exercise, increase the activity of SOD, thereby improving the antioxidant capacity of myocardial tissue, reducing the level of lipid peroxidation, and by downregulating the NF-κB signaling pathway, increasing the antioxidant enzyme activity and anti-inflammatory capacity of myocardial tissue, and reducing myocardial injury caused by exercise exhaustion.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The invention provides a traditional Chinese medicine composition for preventing and treating exercise-induced myocardial injury. The composition is prepared from the following raw materials in parts by weight: 0.8-1.2 parts of total flavonoids of kudzu root, 10-15 parts of kiwi fruit, and 15-20 parts of hawthorn.

[0007] Preferably, the method for extracting total flavonoids from Pueraria lobata comprises the following steps:

[0008] (1) crushing the kudzu root and soaking it in an alkaline solution, and performing microwave extraction to obtain a kudzu root extract;

[0009] (2) adjusting the pH of the Pueraria extract with an acidic solution, and allowing the solution to stand to obtain a product;

[0010] (3) Drying the product to obtain the total flavonoids of Pueraria lobata.

[0011] Preferably, the particle size of the pulverized Pueraria root is 80-120 meshes; the mass volume ratio of the pulverized Pueraria root to the alkaline solution is 1g: (20-30)ml.

[0012] Preferably, the power of the microwave is 200-300W; the microwave time is 5-7min.

[0013] Preferably, the adjusted pH of the Pueraria extract is 4-6; and the standing time is 10-20 minutes.

[0014] Preferably, the alkaline solution includes one or more of sodium hydroxide solution, sodium carbonate solution and potassium carbonate solution, and the pH of the alkaline solution is 8-10; the acidic solution includes one or more of acetic acid solution, hydrochloric acid solution and citric acid solution.

[0015] The present invention also provides a method for preparing the above-mentioned Chinese medicine composition, comprising the following steps:

[0016] (1) respectively extracting kiwi fruit pulp and hawthorn fruit pulp, separating the solid and liquid, and extracting the juice to obtain kiwi fruit juice and hawthorn juice;

[0017] (2) The kiwi juice, hawthorn juice and total flavonoids from kudzu root are mixed, sterilized and homogenized to obtain the traditional Chinese medicine composition.

[0018] Preferably, the sterilization temperature is 80-100°C and the time is 15-20 min; the homogenization pressure is 15-30 MPa, the homogenization temperature is 55-70°C and the time is 2-4 min.

[0019] The present invention also provides the use of the above-mentioned Chinese medicine composition or the Chinese medicine composition obtained by the above-mentioned preparation method in the preparation of a medicine for preventing and treating exercise-induced myocardial injury, wherein the medicine further comprises a pharmaceutically acceptable excipient.

[0020] The present invention provides a Chinese medicine composition for preventing and treating exercise-induced myocardial injury, and a preparation method and application thereof. The total flavonoids of kudzu root prepared by the present invention have higher purity, and are prepared into a Chinese medicine composition by combining it with kiwi fruit and hawthorn, which can reduce the content of MDA in the body during exercise, increase the activity of SOD, and thus improve the antioxidant capacity of myocardial tissue, reduce the level of lipid peroxidation, and improve the antioxidant enzyme activity and anti-inflammatory capacity of myocardial tissue by downregulating the NF-κB signaling pathway, and have a good preventive and therapeutic effect on myocardial injury caused by exhaustive exercise. DETAILED DESCRIPTION

[0021] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention. Example 1

[0022] A traditional Chinese medicine composition for preventing and treating exercise-induced myocardial injury comprises, by weight, 1 part of total flavonoids of kudzu root, 12 parts of kiwi fruit, and 18 parts of hawthorn.

[0023] The preparation process of total flavonoids from Pueraria root is as follows:

[0024] Wash and dry fresh kudzu root, crush it to a particle size of 100 mesh, soak the kudzu root in a sodium hydroxide solution (pH=10) according to the mass volume ratio of kudzu root to sodium hydroxide solution of 1g:25ml, extract for 6min under the action of microwave with power of 250W, and then filter with a 100-mesh filter cloth to obtain kudzu root extract. Adjust the pH of the kudzu root extract to 5 with acetic acid solution, let it stand at 25℃ for 15min, discard the upper solution, and obtain the precipitated product. Freeze-dry the product under vacuum degree of 20Pa and temperature of -80℃ for 12h to obtain total flavonoids of kudzu root. The purity of the total flavonoids of kudzu root was 98.1% as determined by high performance liquid chromatography.

[0025] The preparation process of the Chinese medicine composition is as follows:

[0026] Wash and peel the kiwi fruit to obtain kiwi fruit pulp; wash and pit the hawthorn to obtain hawthorn fruit pulp. Juice the kiwi fruit pulp and hawthorn fruit pulp separately, filter with a 200-mesh filter cloth, and take the juice to obtain kiwi fruit juice and hawthorn juice. After mixing the kiwi fruit juice, hawthorn juice and the above-prepared total flavonoids of kudzu root, sterilize them at 85°C for 20 minutes, and then homogenize them at a pressure of 25MPa and a temperature of 60°C for 3 minutes to make the mixture more uniform, thereby obtaining the traditional Chinese medicine composition. Example 2

[0027] A traditional Chinese medicine composition for preventing and treating exercise-induced myocardial injury comprises, by weight, 1.2 parts of total flavonoids of kudzu root, 15 parts of kiwi fruit, and 15 parts of hawthorn.

[0028] The preparation process of total flavonoids from Pueraria root is as follows:

[0029] Wash and dry fresh kudzu root, crush it to a particle size of 120 mesh, soak the kudzu root in sodium carbonate solution (pH=8) according to the mass volume ratio of kudzu root to sodium carbonate solution of 1g:30ml, extract for 7min under the microwave power of 300W, and then filter with a 100-mesh filter cloth to obtain kudzu root extract. Adjust the pH of the kudzu root extract to 6 with hydrochloric acid solution, let it stand at 25℃ for 20min, discard the upper solution, and obtain the precipitated product. Freeze-dry the product under the conditions of vacuum degree of 15Pa and temperature of -80℃ for 10h to obtain total flavonoids of kudzu root. The purity of the total flavonoids of kudzu root was 97.5% as determined by high performance liquid chromatography.

[0030] The preparation process of the Chinese medicine composition is as follows:

[0031] Wash and peel the kiwi fruit to obtain kiwi fruit pulp; wash and pit the hawthorn to obtain hawthorn fruit pulp. Juice the kiwi fruit pulp and hawthorn fruit pulp separately, filter with a 200-mesh filter cloth, and take the juice to obtain kiwi fruit juice and hawthorn juice. After mixing the kiwi fruit juice, hawthorn juice and the above-prepared total flavonoids of kudzu root, sterilize them at 100° C. for 15 minutes, and then homogenize them at a pressure of 15 MPa and a temperature of 70° C. for 4 minutes to make the mixture more uniform, thereby obtaining the traditional Chinese medicine composition. Example 3

[0032] A traditional Chinese medicine composition for preventing and treating exercise-induced myocardial injury comprises, by weight, 0.8 parts of total flavonoids of kudzu root, 10 parts of kiwi fruit, and 20 parts of hawthorn.

[0033] The preparation process of total flavonoids from Pueraria root is as follows:

[0034] Wash and dry fresh kudzu root, crush it to a particle size of 80 mesh, soak the kudzu root in a potassium carbonate solution (pH=8) according to the mass volume ratio of kudzu root to potassium carbonate solution of 1g:20ml, extract for 7min under the action of microwave with power of 200W, and then filter with a 100-mesh filter cloth to obtain kudzu root extract. The pH of the kudzu root extract was adjusted to 4 with citric acid solution, and it was allowed to stand at 25℃ for 10min, and the upper solution was discarded to obtain a precipitated product. The product was freeze-dried for 16h under the conditions of vacuum degree of 25Pa and temperature of -80℃ to obtain total flavonoids of kudzu root. The purity of the total flavonoids of kudzu root was 95.9% as determined by high performance liquid chromatography.

[0035] The preparation process of the Chinese medicine composition is as follows:

[0036] Wash and peel the kiwi fruit to obtain kiwi fruit pulp; wash and pit the hawthorn to obtain hawthorn fruit pulp. Juice the kiwi fruit pulp and hawthorn fruit pulp separately, filter with a 200-mesh filter cloth, and take the juice to obtain kiwi fruit juice and hawthorn juice. After mixing the kiwi fruit juice, hawthorn juice and the above-prepared total flavonoids of kudzu root, sterilize them at 80°C for 20 minutes, and then homogenize them at a pressure of 30MPa and a temperature of 55°C for 2 minutes to make the mixture more uniform, thereby obtaining the traditional Chinese medicine composition. Comparative Example 1

[0037] Calculated by weight: 1 part of total flavonoids from Pueraria root, 12 parts of kiwi fruit, and 18 parts of hawthorn.

[0038] The preparation process of total flavonoids from Pueraria root is as follows:

[0039] Wash and dry fresh kudzu root, crush it to a particle size of 100 mesh, soak the kudzu root in a sodium hydroxide solution (pH=10) for 6 minutes according to the mass volume ratio of kudzu root to sodium hydroxide solution of 1g:25ml, and then filter it with a 100-mesh filter cloth to obtain a kudzu root extract. Adjust the pH of the kudzu root extract to 5 with acetic acid solution, let it stand at 25°C for 15 minutes, discard the upper solution, and obtain a precipitated product. Freeze-dry the product under vacuum of 20Pa and temperature of -80°C for 12 hours to obtain total flavonoids of kudzu root. The purity of the total flavonoids of kudzu root was 67.2% as determined by high performance liquid chromatography.

[0040] The preparation process of the Chinese medicine composition is as follows:

[0041] Wash and peel the kiwi fruit to obtain kiwi fruit pulp; wash and pit the hawthorn to obtain hawthorn fruit pulp. Juice the kiwi fruit pulp and hawthorn fruit pulp separately, filter with a 200-mesh filter cloth, and take the juice to obtain kiwi fruit juice and hawthorn juice. After mixing the kiwi fruit juice, hawthorn juice and the above-prepared total flavonoids of kudzu root, sterilize them at 85°C for 20 minutes, and then homogenize them at a pressure of 25MPa and a temperature of 60°C for 3 minutes to make the mixture more uniform, thereby obtaining the traditional Chinese medicine composition. Comparative Example 2

[0042] A traditional Chinese medicine composition for preventing and treating exercise-induced myocardial injury comprises, by weight, 1 part of total flavonoids of kudzu root, 12 parts of kiwi fruit, and 18 parts of hawthorn.

[0043] The preparation process of total flavonoids from Pueraria root is as follows:

[0044] Wash and dry fresh kudzu root, crush it to a particle size of 100 mesh, soak the kudzu root in a sodium hydroxide solution (pH=10) according to the mass volume ratio of kudzu root to sodium hydroxide solution of 1g:25ml, extract for 20min under the ultrasonic action of 270W, and then filter with a 100-mesh filter cloth to obtain a kudzu root extract. The pH of the kudzu root extract was adjusted to 5 with acetic acid solution, and it was allowed to stand at 25℃ for 15min, and the upper solution was discarded to obtain a precipitated product. The product was freeze-dried for 12h under the conditions of a vacuum degree of 20Pa and a temperature of -80℃ to obtain total flavonoids of kudzu root. The purity of the total flavonoids of kudzu root was 85.7% as determined by high performance liquid chromatography.

[0045] The preparation process of the Chinese medicine composition is as follows:

[0046] Wash and peel the kiwi fruit to obtain kiwi fruit pulp; wash and pit the hawthorn to obtain hawthorn fruit pulp. Juice the kiwi fruit pulp and hawthorn fruit pulp separately, filter with a 200-mesh filter cloth, and take the juice to obtain kiwi fruit juice and hawthorn juice. After mixing the kiwi fruit juice, hawthorn juice and the above-prepared total flavonoids of kudzu root, sterilize them at 85°C for 20 minutes, and then homogenize them at a pressure of 25MPa and a temperature of 60°C for 3 minutes to make the mixture more uniform, thereby obtaining the traditional Chinese medicine composition.

[0047] Test Example 1

[0048] The preventive and therapeutic effects of the Chinese medicine composition prepared in Example 1 and Comparative Examples 1-2 on exercise-induced myocardial injury in mice were verified. The specific process is as follows:

[0049] 63 mice (weight 20±2g, provided by the Animal Center of Anhui Medical University) were randomly divided into seven groups: normal group, exercise model group, low-dose experimental group (Example 1, 2 ml / 10 g), medium-dose experimental group (Example 1, 5 ml / 10 g), high-dose experimental group (Example 1, 10 ml / 10 g), control experimental group 1 (Comparative Example 1, 2 ml / 10 g), and control experimental group 2 (Comparative Example 2, 2 ml / 10 g). Among them, the normal group and the model group were given blank saline (2 ml / 10 g) by gavage, and each experimental group was given the Chinese medicine composition prepared in Example 1.

[0050] The experimental procedure of exercise-induced myocardial injury in mice is as follows:

[0051] The mice in the model group and each experimental group were trained in swimming without weight for 7 weeks. The training time was concentrated between 9:00 and 11:00 in the morning, and the drug was administered between 6:00 and 8:00 in the afternoon. The training time in the first week was 30 minutes per day, and it was extended by 10 minutes per day in the second week. By analogy, the swimming time in the seventh week was 90 minutes per day. After the last oral administration of the mice, feeding was stopped, and swimming training was performed on the morning of the 50th day until exhaustion. The exhaustion criterion was that the mice could not float on the water surface after 10 seconds in the water, and could not complete the righting reflex when placed on a flat surface.

[0052] The exhausted mice were killed, and myocardial tissue samples and blood samples were collected. CK-MB, MDA, SOD, IL-1β, IL-6, and TNF-α of mice in each group were detected. The test results are shown in Table 1.

[0053] Table 1 Test results of various indicators of mice in each group

[0054] CK-MB(U / L) MDA (nmol / mg) SOD(U / mg) IL-1β (pg / mL) IL-6 (pg / mL) TNF-α(pg / mL) Normal group 521.632±10.542 7.124±0.651 1.218±0.068 135.44±2.641 152.70±1.369 602.4±13.200 Sports Model Group 852.315±16.478 11.524±1.658 0.674±0.136 203.51±1.987 185.36±1.488 768.5±10.541 Low dose experimental group 787.265±20.141 9.415±1.022 0.869±0.054 154.68±2.111 171.44±2.016 726.3±19.514 Medium dose experimental group 652.962±24.215 8.110±1.035 0.967±0.033 136.52±2.063 165.37±2.324 694.1±16.216 High dose experimental group 568.453±18.762 8.021±0.347 0.883±0.041 142.77±1.547 158.33±1.229 652.8±14.856 Control group 1 821.361±13.774 11.514±0.654 0.795±0.062 186.52±1.732 176.54±1.364 765.4±20.154 Control group 2 793.887±20.451 9.847±1.005 0.769±0.064 159.41±2.471 172.36±1.658 732.6±18.577

[0055] As can be seen from Table 1, compared with the normal group, the serum CK-MB in the exercise model group was increased, while that in each experimental group was lower than that in the exercise model group. Among them, the high-dose experimental group was the most significantly reduced, with little difference from the normal group; the control experimental groups 1 and 2 also decreased, but the degree of reduction was smaller than that in the low-dose experimental group.

[0056] Compared with the normal group, the myocardial MDA in the exercise model group increased significantly, while that in each experimental group was lower than that in the exercise model group, among which the high-dose experimental group was the most significantly reduced. There was no significant difference between the control experimental group 1 and the exercise model group, and the control experimental group 2 had a smaller reduction compared with the low-dose experimental group.

[0057] Compared with the normal group, the myocardial SOD in the exercise model group was significantly reduced, while all experimental groups were improved compared with the exercise model group, among which the improvement effect of the medium-dose experimental group was the most obvious; the control experimental groups 1 and 2 also improved, but compared with the low-dose experimental group, the degree of improvement was smaller.

[0058] Compared with the normal group, the serum IL-1β in the exercise model group was increased, while that in each experimental group was lower than that in the exercise model group. Among them, the medium-dose experimental group was the most significantly reduced, with little difference from the normal group; the control experimental groups 1 and 2 also decreased, but the degree of reduction was smaller than that in the low-dose experimental group.

[0059] Compared with the normal group, the serum IL-6 in the exercise model group was increased, while that in all experimental groups was lower than that in the exercise model group. Among them, the high-dose experimental group was the most significantly reduced, with little difference from the normal group; the control experimental groups 1 and 2 also decreased, among which the control experimental group 1 had a smaller decrease compared with the low-dose experimental group, and there was no significant difference between the control experimental group 2 and the low-dose experimental group.

[0060] Compared with the normal group, the serum TNF-α in the exercise model group was increased, while that in each experimental group was lower than that in the exercise model group. Among them, the high-dose experimental group was the most significantly reduced, with little difference from the normal group; the control experimental groups 1 and 2 also decreased, but the degree of reduction was smaller than that in the low-dose experimental group.

[0061] In summary, the Chinese medicine composition of the present invention has obvious preventive and therapeutic effects on exercise-induced myocardial injury in mice.

[0062] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for preventing and treating exercise-induced myocardial injury, characterized in that: The preparation method is prepared from the following raw materials in parts by weight: 0.8-1.2 parts of total flavonoids of kudzu root, 10-15 parts of kiwi fruit, and 15-20 parts of hawthorn; The method for extracting total flavonoids from kudzu root comprises the following steps: (1) crushing the kudzu root and soaking it in an alkaline solution, and performing microwave extraction to obtain a kudzu root extract; (2) adjusting the pH of the kudzu root extract with an acidic solution, and allowing the solution to stand to obtain a product; (3) drying the product to obtain the total flavonoids of Pueraria lobata; The particle size of the pulverized kudzu root is 80-120 meshes; the mass volume ratio of the pulverized kudzu root to the alkaline solution is 1g: (20-30)ml; The power of the microwave is 200-300W; the microwave time is 5-7min; The pH of the Pueraria extract after adjustment is 4-6; the standing time is 10-20 minutes; The alkaline solution includes one or more of a sodium hydroxide solution, a sodium carbonate solution and a potassium carbonate solution, and the pH of the alkaline solution is 8-10; The acidic solution includes one or more of acetic acid solution, hydrochloric acid solution and citric acid solution.

2. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The steps include: (1) respectively extracting kiwi fruit pulp and hawthorn fruit pulp, separating the solid and the liquid, and extracting the juice to obtain kiwi fruit juice and hawthorn juice; (2) The kiwi juice, hawthorn juice and total flavonoids from kudzu root are mixed, sterilized and homogenized to obtain the traditional Chinese medicine composition.

3. The preparation method according to claim 2, characterized in that: The sterilization temperature is 80-100°C and the time is 15-20 minutes; the homogenization pressure is 15-30MPa, the homogenization temperature is 55-70°C and the time is 2-4 minutes.

4. Use of the Chinese medicine composition according to claim 1 or the Chinese medicine composition obtained by the preparation method according to claim 2 or 3 in preparing a drug for preventing and treating exercise-induced myocardial injury.