Sodium Aescinate Alleviated Liver Fibrosis In Rats By Inhibiting The EIF4F Complex Through PI3K/AKT/mTOR Pathway

Object The continuous development process of hepatitis-liver cirrhosis-liver cancer is the general development rule of liver cancer,which is called the trilogy of liver cancer development.As the largest developing country,China is a country with a large number of hepatitis infections and a high incidence of liver cancer patients.The current treatment methods for liver cancer in the world include surgery to remove tumor lesions,B-guided radiofrequency or microwave ablation,radiotherapy,chemical drugs treatment and liver transplantation,etc.But the therapeutic effect is limited.Liver cirrhosis is an important intermediate process in the development of liver cancer,and early intervention to block this process can greatly reduce the possibility of patients developing liver cancer.The development of liver cirrhosis is a long continuous process,and liver fibrosis is the early reversible stage of the development of liver cirrhosis.The main feature of liver fibrosis is that hepatic stellate cells activate and transdifferentiate into myofibroblasts,secreting a large amount of extracellular matrix components including collagen I,collagen III,α-SMA,and fibronectin,deposited in the portal area between the liver lobules.Sodium aescinate is the main ingredient extracted from the traditional Chinese medicine Saluozi.Its known effects include anti-inflammatory,swelling,expansion of blood vessels and improvement of microcirculation,etc.Preliminary experiments have proved that sodium aescinate has a significant inhibitory effect on primary liver cancer.This study mainly explores the effect and mechanism of sodium aescinate on the development of liver fibrosis.Method This study mainly used in vitro and in vivo experiments to determine the effect of sodium aescinate on the development of liver fibrosis.In vivo experiments,male rats were divided into three groups:a blank control group,a CCl4-induced model group,and a treatment group treated with sodium aescinate on the basis of CCl4-induced.The rats in the control group were treated by intraperitoneal injection of olive oil.The rats in the model group and the treatment group were injected with carbon tetrachloride liquid to induce liver fibrosis in the experimental rats.This treatment continued throughout the 8-week experiment.Starting from the third week of the experiment,the treatment group was given an appropriate amount of sodium aescinate for intraperitoneal injection,and the control group and model group were given intraperitoneal injection of the same amount of medical physiological saline.At the end of the 8th week,the rat liver tissues were taken out of the stomach,and the liver fibrosis development degree and the expression of collagen in the tissues of the control group,model group and treatment group were detected by immunohistochemistry,HE and Masson staining.In the in vitro cell test,the growth and proliferation activity of HSC-T6 cells treated with sodium aescinate was detected by MTT and clone formation experiments,and the degree of apoptosis of HSC-T6 cells treated with sodium aescinate was detected by flow cytometry.Western blotting was used to detect the influence of sodium aescinate on the expression of eIF4F complex in HSC-T6 cells and the expression of various proteins in the PI3K/AKT/mTOR pathway.Result CCl₄ induced liver fibrosis in rats significantly.HE,Masson and immunohistochemical staining results showed that sodium aescinate treatment significantly decreased the content ofα-SMA,COL-I and COL-III in the extracellular matrix of rat liver compared with the carbon tetrachloride-induced liver fibrosis model group.MTT test results showed that sodium aescinate significantly reduced the proliferation activity of HSC-T6 cells;clone formation experiment showed that sodium aescinate significantly reduced the number of cell colonies formed in HSC-T6 cells;flow cytometry test results showed that sodium aescinate increased the number of apoptosis in HSC-T6 cells,with the most obvious increase in the number of early apoptosis;Western blotting showed that sodium aescinate can down-regulate the expression ofα-SMA,COL-I and COL-Ⅲin HSC-T6 cells.The results of immunohistochemical staining and western blotting of HSC-T6 cells of rat liver tissue treated with sodium aescinate showed that the expression of e IF4G and e IF4E proteins decreased.Sodium aescinate can down-regulate the expression level of each protein in the PI3K/AKT/mTOR/4EBP1 signaling pathway in HSC-T6 cells,and reduce the phosphorylation level of 4EBP1.Conclusion Sodium aescinate inhibits the synthesis of eIF4F complex through the PI3K/AKT/mTOR pathway to alleviate the progression of liver fibrosis in rats.