Application of Xiaochaihu decoction compound water extract in preparation of medicine and functional food for treating and preventing non-alcoholic steatohepatitis

Abstract

The invention belongs to the technical field of medicines and functional foods, and discloses an application of a Xiaochaihu decoction compound water extract in preparation of medicines and functional foods for treating and preventing non-alcoholic steatohepatitis, the water extract is prepared from the following raw material medicines in parts by weight: radix bupleuri, scutellaria baicalensis, ginseng, pinellia ternate, liquorice and ginger are added into every four Chinese dates; and adding water to soak for 3 hours, adding drinking water for primary decoction, adding drinking water for secondary decoction and adding drinking water for tertiary decoction, filtering the decoctions through a 100-mesh sieve, and combining the decoctions for three times and the concentrated solution obtained by concentrating the water decoction to obtain the common goldenrop herb decoction water extract. The common goldenrop soup has the activity of resisting non-alcoholic steatohepatitis and reducing blood fat, can relieve liver injury, hyperlipidemia and oxidative stress accompanied by NASH patients, can be used for treating the non-alcoholic steatohepatitis and the hyperlipidemia, and can be applied to preparation of medicines and functional foods for treating and preventing the non-alcoholic steatohepatitis and the hyperlipidemia.

Description

Technical Field

[0001] The present invention belongs to the technical field of drugs and functional foods, and in particular relates to the application of a compound aqueous extract of Xiaochaihu Decoction in the preparation of drugs and functional foods for the treatment and prevention of non-alcoholic fatty liver disease. Background Art

[0002] Xiaochaihu Decoction originated from Treatise on Febrile and Miscellaneous Diseases written by Zhang Zhongjing, a medical scientist in the Eastern Han Dynasty. Xiaochaihu Decoction is composed of 24 grams of Bupleuri Radix, 9 grams of Scutellariae Radix, 9 grams of Ginseng Radix, 9 grams of Pinelliae Rhizoma, 6 grams of Glycyrrhizae Radix Preparata, 9 grams of Zingiberis Rhizoma Recens, and 12 pieces of Jujubae Fructus. The Shaoyang syndrome treated by Xiaochaihu Decoction is manifested as alternating chills and fever, fullness and discomfort in the chest and hypochondrium, bitter taste in the mouth and dry throat, dizziness and vertigo, restlessness and preference for vomiting, red tongue with yellow coating, taut and rapid pulse, etc., and has the functions of reconciling shaoyang, replenishing qi and strengthening the spleen, and harmonizing the stomach to stop vomiting. It is a famous representative prescription for reconciling shaoyang in traditional Chinese medicine. Modern pharmacological research has found that Xiaochaihu Decoction has the effects of regulating immunity, anti-inflammation, anti-liver fibrosis, anti-tumor and regulating endocrine, and can be clinically used to treat diseases of the respiratory system, digestive system, immune system, urogenital system and endocrine system.

[0003] Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease at present. The clinical histological phenotype of this disease has extended from non-alcoholic fatty liver (NAFL) to non-alcoholic steatohepatitis (NASH), and the probability of NASH progressing to liver cirrhosis is as high as 15 - 25%. On this basis, NASH is more likely to progress to liver cancer and cause the death of patients. In the past 20 years, a large number of relevant pathological and clinical medical studies aimed at alleviating the development process of such diseases have been carried out all over the world, but no very effective treatment methods have been found yet.

[0004] Through retrieval, no patent publication documents related to the present invention patent application have been found. Summary of the Invention

[0005] The purpose of the present invention is to overcome the problems existing in the prior art and develop the application of a compound aqueous extract of Xiaochaihu Decoction in the preparation of drugs and functional foods for the treatment and prevention of non-alcoholic fatty liver disease.

[0006] The technical solution adopted by the present invention to solve the technical problems is as follows:

[0007] A compound aqueous extract of Xiaochaihu Decoction, wherein the aqueous extract is made from the following raw materials in parts by weight:

[0008] Add 24 grams of Bupleuri Radix, 9 grams of Scutellariae Radix, 9 grams of Ginseng Radix, 9 grams of Pinelliae Rhizoma, 6 grams of Glycyrrhizae Radix Preparata, and 9 grams of Zingiberis Rhizoma Recens to every 4 pieces of Jujubae Fructus;

[0009] Soak in water for 3 hours. For the first decoction, add 10 - 16 times the mass of drinking water; for the second decoction, add 8 - 12 times the mass of drinking water; for the third decoction, add 5 - 8 times the mass of drinking water. The decoction solutions are all filtered through a 100 - mesh sieve, and the three - time decoction solutions are combined. The concentrated solution obtained by concentrating the water - decocted solution is the aqueous extract of Xiaochaihu Decoction.

[0010] Furthermore, the extraction rate of the aqueous extract is 16.36%.

[0011] Furthermore, the aqueous extract can alleviate liver injury caused by non - alcoholic fatty liver disease.

[0012] Furthermore, the aqueous extract has the activity against non - alcoholic fatty liver disease.

[0013] Application of the aqueous extract of Xiaochaihu Decoction compound as described above in the preparation of drugs and / or functional foods for the treatment and / or prevention of non - alcoholic fatty liver disease.

[0014] Advantages and beneficial effects achieved by the present invention:

[0015] Experiments of the present invention have proved that Xiaochaihu Decoction can reduce blood lipid, inflammation and oxidative stress in methionine - choline - deficient (MCD) mice. The present invention has found that the Xiaochaihu Decoction compound has the activities against non - alcoholic fatty hepatitis and reducing blood lipid, can alleviate liver injury and hyperlipidemia accompanied by NASH patients, and can be used for the treatment of non - alcoholic fatty hepatitis and hyperlipidemia, and can be applied to the preparation of drugs and functional foods for the treatment and prevention of non - alcoholic fatty hepatitis and hyperlipidemia. Description of the Drawings

[0016] Figure 1 It is a graph showing the effect of Xiaochaihu Decoction on the body weight change of mice induced by MCD diet in the present invention;

[0017] Figure 2 It is a graph showing the effect of Xiaochaihu Decoction on the body weight of mice on the 28th day induced by MCD diet in the present invention;

[0018] Figure 3 It is a graph showing the effect of Xiaochaihu Decoction on the triglyceride content in the plasma of mice in the present invention;

[0019] Figure 4 It is a graph showing the effect of Xiaochaihu Decoction on the total cholesterol content in the plasma of mice in the present invention;

[0020] Figure 5 It is a graph showing the effect of Xiaochaihu Decoction on the low - density lipoprotein cholesterol content in the plasma of mice in the present invention;

[0021] Figure 6 It is a graph showing the effect of Xiaochaihu Decoction on the high - density lipoprotein cholesterol content in the plasma of mice in the present invention;

[0022] Figure 7It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the activity of glutamic-oxaloacetic transaminase in the plasma of mice;

[0023] Figure 8 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the activity of glutamic-pyruvic transaminase in the plasma of mice;

[0024] Figure 9 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the content of reduced glutathione in the liver tissue of mice;

[0025] Figure 10 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the content of malondialdehyde in the liver tissue of mice;

[0026] Figure 11 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the content of superoxide dismutase in the liver tissue of mice;

[0027] Figure 12 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the pathological changes of HE staining of the liver of mice.

[0028] Figure 13 It is the graph showing the effect of Minor Bupleurum Decoction in the present invention on the pathological changes of oil red O staining of the liver of mice. Detailed implementation mode

[0029] For a better understanding of the present invention, the following further describes the present invention in detail with reference to embodiments, but the scope claimed by the present invention is not limited to the scope shown in the embodiments.

[0030] The raw materials used in the present invention are all conventional commercially available products without special instructions. The methods used in the present invention are all conventional methods in the art, and the masses of various substances used in the present invention are all conventional masses.

[0031] A compound water extract of Minor Bupleurum Decoction, wherein the water extract is prepared from the following raw medicinal materials in parts by weight:

[0032] Add 24 grams of Bupleurum chinense, 9 grams of Scutellaria baicalensis, 9 grams of Ginseng, 9 grams of Pinellia ternata, 6 grams of Licorice, and 9 grams of Fresh Ginger to every 4 Chinese dates; 24 grams of Bupleurum chinense, 9 grams of Scutellaria baicalensis, 9 grams of Ginseng, 9 grams of Pinellia ternata, 6 grams of Licorice, 9 grams of Fresh Ginger, and 4 Chinese dates;

[0033] Soak in water for 3 hours, add 10 - 16 times the mass of drinking water for the first decoction, add 8 - 12 times the mass of drinking water for the second decoction, add 5 - 8 times the mass of drinking water for the third decoction. The decoction solutions are all filtered through a 100 - mesh sieve, and the three decoction solutions are combined. The concentrated solution obtained by concentrating the water decoction solution is the water extract of Minor Bupleurum Decoction.

[0034] Furthermore, the extraction rate of the water extract is 16.36%.

[0035] Furthermore, the water extract can alleviate liver injury caused by non-alcoholic steatohepatitis.

[0036] Furthermore, the water extract has anti-non-alcoholic steatohepatitis activity.

[0037] Use of the water extract of the Xiaochaihu Decoction compound as described above in the preparation of drugs and / or functional foods for the treatment and / or prevention of non-alcoholic steatohepatitis.

[0038] Specifically, the related preparation and detection are as follows:

[0039] A water extract of the Xiaochaihu Decoction compound, wherein the water extract is made from the following raw materials in parts by weight:

[0040] 24 grams of Bupleuri Radix, 9 grams of Scutellariae Radix, 9 grams of Ginseng Radix, 9 grams of Pinelliae Rhizoma, 6 grams of Glycyrrhizae Radix, 9 grams of Zingiberis Rhizoma Recens, 4 pieces of Jujubae Fructus; soaked in water for 3 hours, 10 - 16 times the mass of drinking water is added for the first decoction, 8 - 12 times the mass of drinking water is added for the second decoction, 5 - 8 times the mass of drinking water is added for the third decoction, the decoction liquid is filtered through a 100-mesh sieve, the three decoction liquids are combined, and the concentrated liquid obtained by concentrating the water decoction liquid is the water extract of Xiaochaihu Decoction.

[0041] 1. Anti-non-alcoholic steatohepatitis activity of the water extract of the Xiaochaihu Decoction compound

[0042] 60 male C57BL / 6 mice, 6 weeks old. After 7 days of environmental adaptation, they were randomly divided into a blank group, a model group, a positive drug group, a low-dose Xiaochaihu Decoction group, a medium-dose Xiaochaihu Decoction group, and a high-dose Xiaochaihu Decoction group, with 10 mice in each group.

[0043] The blank group was fed with MCS control feed (TP 3005MS, purchased from Nantong Trofi Co., Ltd.); and at the same time, 0.2 mL of solvent (sodium carboxymethylcellulose with a mass concentration of 0.5%) was given by gavage every day.

[0044] The model group was fed with methionine-choline-deficient (MCD) feed (TP 3005, purchased from Nantong Trofi Co., Ltd.); and at the same time, 0.2 mL of solvent (sodium carboxymethylcellulose with a mass concentration of 0.5%) was given by gavage every day.

[0045] The positive drug group was fed with MCD feed; the clinical drug silymarin was used as a positive control, and the dosing dose was 0.05 g / kg. It was administered by gavage once a day.

[0046] The low-dose Xiaochaihu Decoction group was fed with MCD feed; and at the same time, 15.0 g / kg of the water extract of the Xiaochaihu Decoction compound was given, and it was administered by gavage once a day.

[0047] The medium-dose group of Xiaochaihu Decoction was fed with MCD diet; meanwhile, 30.0 g / kg of the water extract of Xiaochaihu Decoction compound was given by gavage once a day.

[0048] The high-dose group of Xiaochaihu Decoction was fed with MCD diet; meanwhile, 60.0 g / kg of the water extract of Xiaochaihu Decoction compound was given by gavage once a day.

[0049] The experiment lasted for 5 weeks. The first week was the adaptation stage of methionine-choline-deficient diet, and then it was continuously fed for 4 weeks. The mice were weighed every 3 days. Half an hour after the last administration, the mice were sacrificed, and the serum and liver were collected for biochemical index analysis and pathological analysis.

[0050] Methionine is an essential amino acid for the human body, which participates in protein synthesis. It cannot be synthesized in the body and can only be taken in from the outside. When methionine is deficient, the patient's appetite will decrease, and protein synthesis in the body will be blocked, resulting in slow growth and weight loss.

[0051] The following relevant indicators can be detected using existing corresponding commercially available kits, so the specific detection steps do not have to be written out again.

[0052] Figure 1 It shows that after 28 days of feeding, the body weight of the blank group fed with methionine-choline control diet increased slightly compared with that on the 0th day, and the body weight of the other groups showed an obvious downward trend after 28 days of feeding with MCD diet. The body weight of the model group decreased severely to about 14 g, and the body weight of the Xiaochaihu Decoction administration group also showed a downward trend.

[0053] Figure 2 It shows the body weight of the mice on the 28th day of the experiment. The results show that there is a significant difference in body weight between the blank group and the model group and the administration group, proving that the experimental modeling is successful.

[0054] TG, TC, LDL-C, and HDL-C are biomarkers in the clinical diagnosis of NASH. NASH patients are often accompanied by abnormal increases in TG, TC, and LDL-C and abnormal decreases in HDL-C.

[0055] Figure 3It shows the content of triglyceride (TG) in the plasma of mice. In the model group fed with MCD diet for a long time, the TG content of mice increased significantly, reaching 1.57 mmol / L, showing a significant difference compared with the blank group (0.53 mmol / L). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the TG content in the modeled mice decreased significantly, reaching 0.82 mmol / L, 0.90 mmol / L, and 1.00 mmol / L. Its activity in reducing the TG content in the plasma of mice was significantly better than that of the positive drug silymarin (0.85 mmol / L), showing good lipid-lowering activity.

[0056] Figure 4 It shows the content of total cholesterol (TC) in the plasma of mice. In the model group fed with MCD diet for a long time, the TC content of mice increased significantly, reaching 1.29 mmol / L, showing a significant difference compared with the blank group (0.71 mmol / L). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the TC content in the modeled mice decreased significantly, reaching 0.81 mmol / L, 0.87 mmol / L, and 0.98 mmol / L, showing good lipid-lowering activity.

[0057] Figure 5 It shows the content of low-density lipoprotein (LDL) in the plasma of mice. In the model group fed with MCD diet for a long time, the LDL content of mice increased significantly, reaching 0.28 mmol / L, showing a significant difference compared with the blank group (0.09 mmol / L). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the LDL content in the modeled mice decreased significantly, reaching 0.26 mmol / L, 0.28 mmol / L, and 0.28 mmol / L, showing good lipid-lowering activity.

[0058] Figure 6 It shows the content of high-density lipoprotein (HDL) in the plasma of mice. In the model group fed with MCD diet for a long time, the HDL content of mice decreased significantly, reaching 0.21 mmol / L, showing a significant difference compared with the blank group (2.09 mmol / L). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the HDL content in the modeled mice increased significantly, reaching 0.61 mmol / L, 0.43 mmol / L, and 0.28 mmol / L. Its activity in increasing the HDL content in the plasma of mice was significantly better than that of the positive drug silymarin (0.52 mmol / L), showing good lipid-lowering activity.

[0059] In hepatocytes, AST and ALT are mainly present in mitochondria. When the liver undergoes severe lesions, such as necrosis or serious damage, AST and ALT will be released from the cells into the blood, causing a significant increase in the unit activity of AST and ALT in the serum. AST and ALT are important biochemical indicators for detecting hepatitis. When hepatitis occurs, AST and ALT will increase significantly.

[0060] Figure 7 The content of aspartate aminotransferase (AST) in the plasma of mice is shown. In the group of mice fed with the MCD diet for a long time, the AST content increased significantly, reaching 277 Karmen units, showing a significant difference from the blank group (49 Karmen units). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of the extract of Xiao Chai Hu Tang, the AST content in the modeled mice decreased significantly, reaching 138 Karmen units, 172 Karmen units, and 235 Karmen units. Its activity in reducing the AST content in the plasma of mice was significantly better than that of the positive drug silymarin (171 Karmen units), showing good anti-inflammatory activity.

[0061] Figure 8 The content of alanine aminotransferase (ALT) in the plasma of mice is shown. In the group of mice fed with the MCD diet for a long time, the ALT content increased significantly, reaching 352 Karmen units, showing a significant difference from the blank group (49 Karmen units). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of the extract of Xiao Chai Hu Tang, the ALT content in the modeled mice decreased significantly, reaching 132 Karmen units, 173 Karmen units, and 200 Karmen units. Its activity in reducing the ALT content in the plasma of mice was significantly better than that of the positive drug silymarin (206 Karmen units), showing good anti-inflammatory activity.

[0062] Oxidative stress plays a key role in the pathogenesis of many diseases. Lipid peroxidation can cause a large amount of hepatocyte damage, resulting in liver necrosis or liver inflammation. Reduced glutathione (GSH) plays a very important role in antioxidant defense, nutrient metabolism, and regulation of cellular events. The liver is the main synthesis site of GSH, and GSH in the liver can resist lipid peroxidation. Malondialdehyde (MDA) is one of the products of lipid peroxidation and is generally used as a marker for indirectly measuring oxidative damage. Superoxide dismutase (SOD) is an important protective substance in the liver. SOD has special physiological activity and is the primary substance for scavenging free radicals in the body.

[0063] Figure 9It shows the content of reduced glutathione (GSH) in the liver tissue of mice. In the model group of mice fed with MCD diet for a long time, the GSH content decreased significantly, reaching 2.82 μmol / gprot, showing a significant difference compared with the blank group (16.31 μmol / gprot). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the GSH content in the modeled mice increased significantly, reaching 14.17 μmol / gprot, 10.69 μmol / gprot, and 6.63 μmol / gprot. Its activity in increasing the GSH content in mouse plasma was significantly better than that of the positive drug silymarin (4.07 μmol / gprot), showing good antioxidant activity.

[0064] Figure 10 It shows the content of malondialdehyde (MDA) in the liver tissue of mice. In the model group of mice fed with MCD diet for a long time, the MDA content increased significantly, reaching 6.27 nmol / mgprot, showing a significant difference compared with the blank group (1.13 nmol / mgprot). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the MDA content in the liver tissue of the modeled mice decreased significantly, reaching 2.11 nmol / mgprot, 2.84 nmol / mgprot, and 3.56 nmol / mgprot. Its activity in decreasing the MDA content in mouse liver tissue was significantly better than that of the positive drug silymarin (2.93 nmol / mgprot), showing good antioxidant activity.

[0065] Figure 11 It shows the content of superoxide dismutase (SOD) in the liver tissue of mice. In the model group of mice fed with MCD diet for a long time, the SOD content decreased significantly, reaching 342 U / mgprot, showing a significant difference compared with the blank group (618 U / mgprot). After administration of high (60 g / kg), medium (30 g / kg), and low (15 g / kg) doses of Bupleurum chinense decoction extract, the SOD content in the liver tissue of the modeled mice increased significantly, reaching 506 U / mgprot, 494 U / mgprot, and 466 U / mgprot, showing good antioxidant activity.

[0066] Figure 12It shows the pathological changes of HE staining in the livers of mice. After induction with a methionine-choline-deficient diet, fatty vacuoles, diffuse steatosis, ballooning degeneration of hepatocytes, and loose and pale staining of cytoplasm due to NASH liver injury appeared in the livers of mice in the model group. After treatment with the positive drug silymarin, the pathological changes in the liver were significantly alleviated. The treatment with Xiao Chai Hu Tang showed good anti-non-alcoholic steatohepatitis activity. The fatty vacuoles in the figure were significantly reduced, indicating a decrease in the degree of lipid accumulation in the livers of mice, which was consistent with the results shown by the biochemical indicators.

[0067] Figure 13 It shows the pathological changes of Oil Red O staining in the livers of mice. After induction with a methionine-choline-deficient diet, a large amount of fat accumulation appeared in the livers of mice in the model group, which was stained red by Oil Red O. After treatment with the positive drug silymarin, the fat accumulation was significantly improved, and the lipids in the liver were significantly reduced. After treatment with the compound extract of Xiao Chai Hu Tang, the red area stained by Oil Red O in the livers of mice was significantly reduced, and the fat accumulation was significantly improved, which was consistent with the test results of the biochemical indicators.

[0068] In summary, the compound of Xiao Chai Hu Tang has good anti-non-alcoholic steatohepatitis activity and has broad prospects in the development and application of drugs for the treatment and prevention of non-alcoholic steatohepatitis and lipid-lowering functional foods. The compound of Xiao Chai Hu Tang of the present invention can be used in the preparation of drugs for the treatment and prevention of non-alcoholic steatohepatitis, and can also be used as a food composition for the preparation of lipid-lowering functional foods, including but not limited to its application in the treatment of non-alcoholic steatohepatitis.

[0069] Although the embodiments of the present invention are disclosed for illustrative purposes, those skilled in the art can understand that various substitutions, changes, and modifications are possible without departing from the spirit and scope of the present invention and the appended claims. Therefore, the scope of the present invention is not limited to the content disclosed in the embodiments.

Claims

1. A water extract of Xiao Chaihu Decoction compound, characterized in that: The water extract is made from the following raw materials in parts by weight: For every 4 jujubes, add 24g of Bupleurum, 9g of Scutellaria, 9g of Ginseng, 9g of Pinellia, 6g of Licorice, and 9g of Ginger; Soak in water for 3 hours, add 10-16 times the mass of drinking water for the first decoction, 8-12 times the mass of drinking water for the second decoction, and 5-8 times the mass of drinking water for the third decoction. The decoctions are all filtered through a 100-mesh sieve, and the three decoctions are combined. The concentrated solution of the decoctions is the Xiao Chaihu Decoction water extract.

2. The Xiao Chai Hu Tang compound water extract according to claim 1, characterized in that: The extraction rate of the water extract was 16.36%.

3. The Xiao Chai Hu Tang compound water extract according to claim 1, characterized in that: The water extract can alleviate liver damage caused by non-alcoholic steatohepatitis.

4. The Xiao Chai Hu Tang compound water extract according to any one of claims 1 to 3, characterized in that: The water extract has anti-nonalcoholic fatty hepatitis activity.

5. Use of the Xiao Chaihu Decoction compound water extract according to any one of claims 1 to 4 in the preparation of medicines and / or functional foods for treating and / or preventing non-alcoholic fatty hepatitis.

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