Application of eleutheroside E in preparation of medicine for preventing or treating hepatic fibrosis

Abstract

The invention discloses application of eleutheroside E in preparation of a medicine for preventing or treating hepatic fibrosis, and belongs to the technical field of biological medicine. The chemical name of the eleutheroside E is 4-[(1R, 3aS, 4S, 6aR)-4-[4-(beta-D-glucoside oxygen group)-3, 5-dimethoxy phenyl] tetrahydro-1H, 3H-furo [3, 4-c] furan-1-yl]-2, 6-dimethoxy phenyl-beta-D-glucoside, and the molecular formula of the eleutheroside E is C34H46O18. Researches find that the eleutheroside E has a remarkable effect of preventing or treating hepatic fibrosis diseases, does not have remarkable hepatotoxicity, and can be used for preparing the medicine for preventing or treating hepatic fibrosis. Cell experiment results show that the eleutheroside E can significantly reduce the proliferation activity of mouse hepatic stellate cells JS-1 and inhibit hepatic stellate cell activation, presents dose dependence and can be used for preparing the medicine for preventing or treating hepatic fibrosis.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of eleutheroside E in the preparation of a drug for preventing or treating liver fibrosis. Background Art

[0002] Liver fibrosis is a pathological process in which the liver, under repeated stimulation by chronic injuries (such as viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease, etc.), shows abnormal deposition of extracellular matrix due to excessive repair responses. Pathologically, it is mainly manifested as the proliferation of intrahepatic collagen fibers, the disorder of hepatic lobule structure, and may be accompanied by the formation of pseudolobules. In severe cases, it can progress to liver cirrhosis. Epidemiological data show that about 25% - 30% of chronic liver disease patients globally will develop liver fibrosis. Among them, viral hepatitis (especially hepatitis B and C) is highly prevalent in Asia and Africa, while alcoholic liver disease and non-alcoholic fatty liver disease are more common in developed countries. Early liver fibrosis is reversible, but if not intervened in time, it may cause portal hypertension, liver failure or liver cancer, which is a key prevention and treatment stage in the management of chronic liver diseases. Summary of the Invention

[0003] The purpose of the present invention is to provide the application of eleutheroside E in the preparation of a drug for treating liver diseases.

[0004] The present invention provides the application of eleutheroside E or its pharmaceutically acceptable salt in the preparation of a drug for preventing or treating liver fibrosis.

[0005] The chemical name of eleutheroside E is 4 - [(1R,3aS,4S,6aR)-4-[4-(β-D-glucopyranosyloxy)-3,5-dimethoxyphenyl]tetrahydro-1H,3H-furo[3,4-c]furan-1-yl]-2,6-dimethoxyphenyl-β-D-glucopyranoside, and its molecular formula is C 34 H 46 O 18 , and its structural formula is as follows:

[0006]

[0007] Further, the drug is used to inhibit the cell proliferation activity of hepatic stellate cells.

[0008] Further, the liver fibrosis is liver fibrosis caused by excessive proliferation and activation of hepatic stellate cells.

[0009] Further, the drug is used to inhibit the activation of hepatic stellate cells.

[0010] Further, the drug refers to eleutheroside E or its pharmaceutically acceptable salt as a single active ingredient.

[0011] Further, the drug is a tablet, granule, pill, capsule or injection.

[0012] The proliferation and activation of hepatic stellate cells (HSCs) are the core links in the occurrence and development of liver fibrosis. In normal liver, HSCs are in a quiescent state, mainly storing vitamin A and maintaining the dynamic balance of the extracellular matrix. When the liver is chronically damaged, HSCs are activated and transformed into myofibroblast-like cells, with enhanced proliferation ability and extracellular matrix synthesis ability. The present invention starts from the mechanism of action of the proliferation and activation of hepatic stellate cells in liver fibrosis, and studies the pharmacological effects of eleutheroside E as a drug for preparing anti-liver fibrosis drugs.

[0013] Compared with the prior art, the present invention provides the use of eleutheroside E or its pharmaceutically acceptable salts in the preparation of drugs for preventing or treating liver fibrosis. It has been found that eleutheroside E has a significant effect on preventing or treating liver fibrosis diseases, and has no significant hepatotoxicity, and can be used to prepare drugs for preventing or treating liver fibrosis. The results of cell experiments show that eleutheroside E can significantly reduce the proliferation activity of mouse hepatic stellate cell line JS-1, inhibit the activation of hepatic stellate cells, and show a dose-dependent manner. Description of the Drawings

[0014] Figure 1 It is a bar graph of the proliferation activity of hepatocytes AML-12 in each group of mice in Example 1;

[0015] Figure 2 It is a bar graph of the proliferation activity of mouse hepatic stellate cell line JS-1 in each group of mice in Example 2, where * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 vs. the control group;

[0016] Figure 3 It is the effect on the expression of activation markers α-SMA and Fibronectin proteins of mouse hepatic stellate cell line JS-1 in each group of mice in Example 3;

[0017] Figure 4 It is the relative expression levels of activation markers α-SMA and Fibronectin proteins of mouse hepatic stellate cell line JS-1 in each group of mice in Example 3, where * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. the control group. Detailed Embodiments

[0018] Combined with the embodiments of the present invention, the technical solutions in the embodiments of the present invention are clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope of protection of the present invention.

[0019] The present invention provides a potential anti-hepatic fibrosis drug, and the following further describes the specific implementation manners of the present invention in detail.

[0020] Example 1

[0021] Effect of eleutheroside E on the proliferation activity of mouse hepatocytes AML-12.

[0022] Hepatocytes in the logarithmic growth phase were collected, the cell suspension concentration was adjusted, and inoculated into 96-well plates, 180 μL per well, about 1×10 4 cells per well. Cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C and 5% CO2 for 24 h until the cells adhered. The cells were divided into 5 groups. Except for the normal control group treated with an equal volume of solvent, the remaining 4 groups of cells were treated with eleutheroside E (25, 50, 100, and 200 μM) for 24 h. Each group had 12 replicate wells. After the drug treatment, 10 μL of CCK-8 solution was added to each well and continued to be cultured for 4 hours, and the absorbance value was measured at 450 nm on an enzyme-linked immunosorbent detector.

[0023] As Figure 1 shown, 25, 50, 100, and 200 μM eleutheroside E had no significant effect on the proliferation activity of mouse hepatocytes AML-12. It indicates that eleutheroside E at the said concentration has no obvious hepatotoxicity.

[0024] Example 2

[0025] Effect of eleutheroside E on the proliferation activity of mouse hepatic stellate cells JS-1.

[0026] Hepatic stellate cells in the logarithmic growth phase were collected, the cell suspension concentration was adjusted, and inoculated into 96-well plates, 180 μL per well, about 1×10 4Cells. They were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C and 5% CO2 for 24 h until the cells adhered. The cells were divided into 5 groups. Except for the normal control group treated with an equal volume of solvent, the remaining 4 groups of cells were treated with eleutheroside E (25, 50, 100, and 200 μM) for 24 h. Each group had 12 replicate wells. After the drug treatment, 10 μL of CCK-8 solution was added to each well and cultured for another 4 hours, and the absorbance value was measured at 450 nm using an enzyme-linked immunosorbent assay detector.

[0027] As Figure 2 shown, 25, 50, 100, and 200 μM of eleutheroside E inhibited the proliferation activity of mouse hepatic stellate cells JS-1, and showed an obvious concentration dependence. It indicates that eleutheroside E dose-dependently inhibits the proliferation activity of hepatic stellate cells.

[0028] Example 3

[0029] Effect of eleutheroside E on the protein expression of activation markers α-SMA and Fibronectin in mouse hepatic stellate cells JS-1.

[0030] Hepatic stellate cells in the logarithmic growth phase were collected, the cell suspension concentration was adjusted, and 2 mL was inoculated into each well of a six-well plate, about 5×10 5 cells per well. They were cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C and 5% CO2 for 24 h until the cells adhered. When the cell density reached 50%, the cells were divided into 4 groups. Except for the normal control group treated with an equal volume of solvent, the remaining 3 groups of cells were treated with eleutheroside E (25, 50, and 100 μM) for 24 h. The medium was discarded, and the cells were washed 3 times with pre-cooled phosphate buffer. Total cell protein was extracted, and the expression levels of the activation markers α-SMA and Fibronectin proteins in hepatic stellate cells were detected by immunoblotting. The results are as Figure 3 shown.

[0031] Figure 4 For the relative expression levels of the activation markers α-SMA and Fibronectin proteins in mouse hepatic stellate cells JS-1 in each group in Example 3, the left figure is the relative expression level of the activation marker α-SMA, and the right figure is the relative expression level of the Fibronectin protein. According to Figure 3 and Figure 4 it can be seen that eleutheroside E reduces the expression of α-SMA and Fibronectin proteins in hepatic stellate cells, and the effect is enhanced with the increase of concentration. It indicates that eleutheroside E dose-dependently and significantly inhibits the activation of hepatic stellate cells.

[0032] The above results indicate that eleutheroside E significantly inhibits the activation of mouse hepatic stellate cells and has good anti-hepatic fibrosis ability.

[0033] The above description is only a preferred embodiment of the present invention and does not impose any formal limitations on the present invention. Although the present invention has been disclosed above with the preferred embodiment, it is not intended to limit the present invention. Any person skilled in the art of this patent can make some changes or modifications to equivalent embodiments with equivalent changes by using the technical content prompted above within the scope of the technical solution of the present invention. However, as long as the content does not depart from the technical solution of the present invention, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention still fall within the scope of the present invention.

Claims

1. Use of eleutheroside E or a medicinal salt thereof in the preparation of a drug for preventing or treating liver fibrosis.

2. The application according to claim 1, wherein The drug is used to inhibit the cell proliferation activity of hepatic stellate cells.

3. The application according to claim 1, wherein The liver fibrosis is liver fibrosis caused by excessive proliferation and activation of hepatic stellate cells.

4. The application according to claim 1, wherein The drug is used to inhibit the activation of hepatic stellate cells.

5. The application according to claim 1, characterized in that The drug refers to eleutheroside E or a medicinal salt thereof as a single active ingredient.

6. The application according to claim 1, characterized in that, The drug is a tablet, granule, pill, capsule or injection.

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