Sodium Aescinate Retards Liver Fibrosis By Inhibiting Phosphorylation Of 4EBP1

Objective Liver fibrosis is a pathological process caused by acute or chronic liver injury,which can progress to cirrhosis,causing serious complications such as liver failure and hepatocellular carcinoma.A large number of deaths each year due to cirrhosis and hepatocellular carcinoma brings a heavy economic burden on many countries.At present,there is no good treatment for liver fibrosis,but the pace of finding potential liver fibrosis treatment drugs has never stopped.This topic mainly studies the role of sodium aescinate in experimental liver fibrosis and its effect on rat hepatic stellate cells,and explores the possible related mechanisms.It provides a theoretical basis for the potential clinical application value of sodium aescinate in liver fibrosis.Methods?1?Forty-one healthy male SD rats were randomly divided into normal control group?n=5?,model group?n=18?,and sodium aescinate treatment group?n=18?.The model group and sodium aescinate treatment group were injected intraperitoneally with a 50%CCl₄ olive oil solution twice a week for a total of 8 weeks.The normal control group was given the same dose of olive oil.In the sodium treatment group,from the 3rd week,3.6 mg·kg^-1 sodium aescinate solution was intraperitoneally injected once a day for 6 weeks.The normal control group and the model group were given the corresponding physiological saline solution at the same time.?2?HE staining was used to observe the morphology and structure of rat liver tissues,and Masson staining was used to observe the proliferation of collagen fibers.?3?Immunohistochemical staining was used to detect the expression of p-4EBP1,?-SMA,COL-?and COL-?in rat liver tissues.?4?Culture of rat HSC-T6 cells.?5?MTT method was used to detect the effect of sodium aescinate on the proliferation of HSC-T6 cells.?6?PI staining method was used to detect the effect of sodium aescinate on cell cycle of HSC-T6 cells.?7?Annexin V-FITC/PI double staining method was used to detect the effect of sodium aescinate on apoptosis of HSC-T6 cells.?8?Western blot was used to detect the effects of sodium aescinate on p-4EBP1,?-SMA,COL-?and COL-?protein expression in HSC-T6 cells.Results?1?The rat liver fibrosis model was successfully constructed,and sodium aescinate could improve the degree of rat liver fibrosis induced by CCl₄.?2?Both HE and Masson staining showed that sodium aescinate can reduce the production of collagen fibers induced by CCl₄.?3?Immunohistochemical staining showed that sodium aescinate could reduce the expression of p-4EBP1,COL-?and COL-?,but had no significant effect on the expression of?-SMA.?4?MTT results showed that sodium aescinate treatment could significantly inhibit the proliferation of HSC-T6 cells for 48 hours.?5?PI staining test showed that 30?mol·L^-1 of sodium aescinate treatment for 48 hours had no significant effect on cell cycle of HSC-T6 cells.?6?The results of Annexin V-FITC/PI double staining showed that sodium aescinate could significantly promote the early apoptosis of HSC-T6 cells under the same conditions.?7?Western blot results showed that after 48 hours treatment of 30?mol·L^-1 of sodium aescinate,the expression of p-4EBP1,COL-?,and COL-?in HSC-T6 cells can be significantly reduced,while there was no significant effect on the expression of?-SMA.Conclusions Sodium aescinate can inhibit the proliferation of hepatic stellate cells,promote its apoptosis,reduce the production of collagen,and then slow down the process of liver fibrosis.The related mechanism may be related to the inhibition of 4EBP1phosphorylation by sodium aescinate.