Traditional Chinese medicine composition for preventing or treating hepatic fibrosis

Abstract

The invention provides a traditional Chinese medicine composition for preventing or treating hepatic fibrosis. The traditional Chinese medicine composition contains total salvianolic acid and total triterpene acid of hawthorn. When the traditional Chinese medicine composition is used for preventing or treating hepatic fibrosis, the hepatic fibrosis area can be reduced, and the pathological score of hepatic fibrosis can be improved. The traditional Chinese medicine composition can also improve hepatic fibrosis by reducing the content of liver hydroxyproline.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicines and relates to a traditional Chinese medicine composition. The traditional Chinese medicine composition contains total phenolic acids from salvia miltiorrhiza and total triterpenic acids from hawthorn. The traditional Chinese medicine composition can be used to prevent or treat liver fibrosis. Background Art

[0002] Liver fibrosis is a common pathological stage in the development of many chronic liver diseases. It is a pathological process in which a variety of pathogenic factors lead to abnormal growth of intrahepatic connective tissue and excessive accumulation of diffuse extracellular matrix (ECM) in the liver. If timely and effective treatment is not taken, it will lead to cirrhosis and liver cancer.

[0003] Patent CN02112488.4 discloses a pharmaceutical composition containing total phenolic acids from salvia miltiorrhiza and total triterpenic acids from hawthorn, which is used to prevent or treat hyperlipidemia. Document 1 discloses a study on the prevention of hyperlipidemia in rats by extracts from salvia miltiorrhiza and hawthorn. Rats developed hyperlipidemia and hepatic steatosis after oral administration of fat emulsion for 14 days, and administration of salvia miltiorrhiza and hawthorn extract inhibited the formation of hyperlipidemia and hepatic steatosis.

[0004] At present, the prior art has not reported that the Chinese medicine composition containing total phenolic acids of Salvia miltiorrhiza and total triterpenic acids of Crataegus pinnatifida can be used for preventing or treating liver fibrosis.

[0005] Reference 1: Journal of Integrated Traditional Chinese and Western Medicine for Cardiovascular and Cerebrovascular Diseases, 2015, 13(11): 3. Summary of the invention

[0006] The invention provides a traditional Chinese medicine composition, which contains total phenolic acids from salvia miltiorrhiza and total triterpenic acids from hawthorn. The traditional Chinese medicine composition is used for preventing or treating liver fibrosis.

[0007] In the Chinese medicine composition, the content of total phenolic acids of Salvia miltiorrhiza accounts for 80-20% of the total weight of the Chinese medicine composition, and the content of total triterpene acids of Crataegus pinnatifida accounts for 20-80% of the total weight of the Chinese medicine composition.

[0008] The traditional Chinese medicine composition of the present invention improves liver fibrosis by reducing the hydroxyproline content in the liver.

[0009] The Chinese medicine composition of the present invention further contains one or more pharmaceutically acceptable excipients.

[0010] Preferably, the pharmaceutically acceptable excipients include one or more of a diluent, an antioxidant, a preservative, a solubilizer, a disintegrant, a lubricant, a pH regulator, a colorant, a dispersant, an osmotic pressure regulator or a surfactant.

[0011] Preferably, the diluent is selected from one or more of sucrose, lactose, galactose, glucose, starch, dextrin, gelatin, microcrystalline cellulose, carboxymethyl cellulose, ethyl cellulose and the like.

[0012] Preferably, the antioxidant is selected from one or more of ascorbic acid or butylated hydroxytoluene (BHT).

[0013] Preferably, the preservative is selected from one or more of benzoic acid, sodium benzoate, p-hydroxybenzoate, sorbic acid or glycerin.

[0014] Preferably, the lubricant is selected from one or more of magnesium stearate, calcium stearate or talc.

[0015] The Chinese medicine composition of the present invention is selected from capsules, tablets, granules, pills, powders, dispersants, solutions, emulsions or suspensions, etc.; preferably, the Chinese medicine composition is selected from capsules or tablets.

[0016] The dosage of the traditional Chinese medicine composition of the present invention is 0.1-1000 mg / kg / day, preferably 1-1000 mg / kg / day, and more preferably 1-500 mg / kg / day.

[0017] The dosage of the traditional Chinese medicine composition of the present invention is 250-5000 mg / day, preferably 500-3750 mg / day, and more preferably 750-2500 mg / day.

[0018] The traditional Chinese medicine composition of the invention is a capsule for eliminating stasis and reducing fat.

[0019] The Xiaoyujiangzhi capsule is used for preventing or treating liver fibrosis.

[0020] The second aspect of the present invention provides a use of any of the above-mentioned Chinese medicine compositions in the preparation of a medicament for preventing or treating liver fibrosis.

[0021] Use of Xiaoyujiangzhi Capsule in preparing medicine for preventing or treating liver fibrosis.

[0022] The Chinese medicine composition of the present invention can reduce the area of ​​liver fibrosis and improve the pathological score of liver fibrosis in the prevention or treatment of liver fibrosis. The Chinese medicine composition can also improve liver fibrosis by reducing the content of hydroxyproline in the liver. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Effects of long-term administration of the Chinese medicine composition XYJZ on liver tissue fibrosis in C57BL / 6J mice (Note: *, compared with Vehicle, P<0.05; ***, compared with Vehicle, P<0.001) DETAILED DESCRIPTION

[0024] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0025] In the following examples, unless otherwise specified, the reagents and consumables used were purchased from conventional reagent manufacturers in the field; unless otherwise specified, the experimental methods and technical means used were conventional methods and means in the field.

[0026] Example 1

[0027] Long-term administration of a traditional Chinese medicine composition (XYJZ) improves carbon tetrachloride-induced liver fibrosis in C57BL / 6J mice

[0028] Preparation of Chinese medicine composition:

[0029] The Chinese medicine composition was prepared according to patent CN 02112488.4 and is hereinafter referred to as XYJZ.

[0030] Animal modeling:

[0031] The carbon tetrachloride (CCl4)-induced liver fibrosis model of C57BL / 6J mice was used to observe the improvement effect of long-term oral administration of a traditional Chinese medicine composition (XYJZ) once a day on liver fibrosis caused by CCl4 in mice.

[0032] 72 male C57BL / 6J mice, 6-7 weeks old, were fed adaptively for about 2 weeks before the experiment. All C57BL / 6J mice were randomly weighed on the morning of the modeling day and injected intraperitoneally with 10% CCl4 or solvent olive oil according to body weight (40 μL for 17.5 g to 22.5 g, 50 μL for 22.5 g to 27.5 g, and 60 μL for 27.5 g to 32.5 g). The modeling was done three times a week and the mice were randomly divided into 6 groups according to body weight, with 12 mice in each group. They were the model control group (Vehicle), 1 positive control drug group (OCA, obeticholic acid) and 3 doses of XYJZ groups; C57BL / 6J mice in the olive oil injection group were used as the normal control group (NC). Drug administration began on the second day after grouping. The specific grouping is shown in Table 1.

[0033] Dosage regimen:

[0034] Preparation method:

[0035] Preparation of XYJZ: Accurately weigh the Chinese medicine composition, add the corresponding volume of sterile water, sonicate until it is completely dissolved into a clear solution, and administer the medicine once a day.

[0036] Preparation of OCA: Accurately weigh the drug in a mortar and grind it thoroughly. First, add 0.5% CMC-Na solution, grind it thoroughly to dissolve, and then make up to the corresponding volume. Dispense the drug once a week.

[0037] Oral administration, the dosage volume is 10 mL / kg, once a day, the administration time is the morning of the day of administration.

[0038] Table 1 Grouping of experimental animals

[0039]

[0040] Detection indicators:

[0041] 1. Random body weight: The random body weight of mice in each group was measured three times a week, on the morning of the day of modeling.

[0042] 2. Food intake: The food intake of each group of mice was measured three times a week, in the morning of the day of modeling.

[0043] 3. Determination of plasma liver indexes AST and ALT. At the 4th and 7th week of administration, blood was collected from the retro-orbital venous plexus of each group of animals after isoflurane anesthesia, and plasma was obtained by centrifugation and frozen at -80°C for the determination of plasma AST and ALT levels.

[0044] 4. Dissection: On the day of dissection, blood was collected from the retro-orbital venous plexus of each group of animals after isoflurane anesthesia, and plasma was centrifuged and frozen at -80℃ for testing of plasma biochemical indicators; then the animals were killed by cervical dislocation, and the liver tissue was separated and weighed, and its ratio to body weight was calculated. At the same time, the liver tissue was fixed with 4% paraformaldehyde, and part of the tissue was frozen in liquid nitrogen.

[0045] 5. Statistics of fibrosis area: After dissection of each group of animals, liver tissue sections fixed with 4% paraformaldehyde were stained with picrosirius red, photographed, and the proportion of fibrosis area was calculated.

[0046] 6. Detection of hydroxyproline content in liver tissue.

[0047] (1) Tissue hydrolysis: weigh about 100 mg of frozen liver tissue into a 10 mL centrifuge tube, add 1 mL of hydrolysis solution, and hydrolyze in a 95°C water bath for 20 min; then adjust the pH to about 6.0-6.8;

[0048] (2) Sample detection: Add an appropriate amount of activated carbon (about 20-30 mg, the supernatant is considered clear and colorless after centrifugation) to the hydrolyzate, centrifuge at 3500 rpm for 10 minutes, take 1 mL of the supernatant, and perform the detection according to the steps in the commercial instructions (Nanjing Jiancheng Bioengineering Research Institute). Calculate the hydroxyproline content based on the absorbance value of each well at a wavelength of 550 nm detected by an ELISA reader.

[0049] 7. Scoring criteria for fibrosis: The scoring criteria for fibrosis are based on the ISHAK scoring criteria. The Sirius red-stained pathological films are scored as follows:

[0050] (1) 0 points: no fibrosis;

[0051] (2) 1. Partial portal fibrosis with or without short fibrous septa;

[0052] (3) 2 points: fibrosis and enlargement of most portal areas, with or without short fibrous septa;

[0053] (4) 3 points: most of the portal tract fibrosis is enlarged, and portal tract-portal tract bridging fibrosis is occasionally seen;

[0054] (5) 4 points: portal area fibrosis is enlarged, accompanied by obvious bridging fibrosis (including portal area-portal area; portal area-central vein);

[0055] (6) 5 points: significant bridging fibrosis (including portal area-portal area and portal area-central vein), with occasional regenerative nodules (incomplete cirrhosis);

[0056] (7) 6 points: possible or definite cirrhosis.

[0057] Data processing and statistical analysis:

[0058] Data are presented as mean ± standard error ( ) indicates that the data were statistically analyzed using Student-t test, p < 0.05 was considered statistically significant, and T-Test was one-tailed. All data are presented in tables, and some data are also presented in graphs.

[0059] Experimental results:

[0060] 1. Effects of long-term administration of the traditional Chinese medicine composition XYJZ on liver fibrosis in C57BL / 6J mice

[0061] According to the Ishak scoring standard, the pathological scoring of the Sirius red staining results showed that compared with the normal control group, the pathological score of the CCL4 modeling group was 4 points, indicating that the liver fibrosis of the mice was significant. The pathological score of the low-dose group of the Chinese medicine composition XYJZ was close to 4 points, and there was no significant difference from the model control group; after long-term administration of the medium-dose group and the high-dose group of XYJZ, the fibrosis score was significantly reduced, indicating that the fibrosis in this dose group was effectively improved (Table 2). Further, we conducted statistics on Sirius red staining. The results showed that although there was no significant difference in the pathological score of the low-dose group of the Chinese medicine composition XYJZ, the area of ​​collagen stained with Sirius red had decreased significantly. The medium-dose group, high-dose group and positive OCA dose group of XYJZ can significantly reduce the area of ​​liver fibrosis (Table 2,Figure 1 ). The above results indicate that XYJZ administration can significantly improve CCL4-induced liver fibrosis in mice.

[0062] Table 2 Effects of the Chinese medicine composition XYJZ on liver fibrosis in C57BL / 6J mice

[0063]

[0064] 2. Effect of long-term administration of the Chinese medicine composition XYJZ on the hydroxyproline content in the liver of C57BL / 6J mice

[0065] Hydroxyproline is a collagen-specific amino acid. In order to further directly reflect the reduction of collagen fibers, we further detected the content of hydroxyproline in the liver of each group of mice. The results showed that compared with the model control group, XYJZ 1000 mg / kg and positive OCA dose groups could significantly reduce the content of hydroxyproline in the liver, and each dose group of XYJZ had a certain dose dependence, as shown in Table 3.

[0066] Table 3 Effects of the Chinese medicine composition XYJZ on the hydroxyproline content in the liver of C57BL / 6J mice

[0067]

Claims

1. A Chinese medicine composition, comprising total phenolic acids from Salvia miltiorrhiza and total triterpenic acids from Crataegus pinnatifida, and is used for preventing or treating liver fibrosis.

2. The Chinese medicine composition according to claim 1, in, The content of total phenolic acid of Salvia miltiorrhiza accounts for 80-20% of the total weight of the traditional Chinese medicine composition, and the content of total triterpene acid of Crataegus pinnatifida accounts for 20-80% of the total weight of the traditional Chinese medicine composition.

3. The Chinese medicine composition according to claim 1, which improves liver fibrosis by reducing liver hydroxyproline content.

4. The Chinese medicine composition according to claim 1, further comprising one or more pharmaceutically acceptable excipients.

5. The Chinese medicine composition according to claim 1, wherein the Chinese medicine composition is selected from capsules, tablets, granules, pills, powders, dispersants, solutions, emulsions or suspensions; preferably capsules or tablets.

6. The Chinese medicine composition according to claim 1, wherein the dosage of the Chinese medicine composition is 0.1-1000 mg / kg / day, preferably 1-1000 mg / kg / day, and more preferably 1-500 mg / kg / day.

7. The Chinese medicine composition according to claim 1, wherein the dosage of the Chinese medicine composition is 250-5000 mg / day, preferably 500-3750 mg / day, and more preferably 750-2500 mg / day. The Chinese medicine composition according to claim 1 , which is Xiaoyu Jiangzhi Capsule.

9. Use of the traditional Chinese medicine composition according to any one of claims 1 to 8 in the preparation of a medicament for preventing or treating liver fibrosis.

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