Novel Matrine Derivative Inhibits The Activation Of Hepatic Stellate Cells And Hepatic Fibrogenesis

Hepatic fibrosis is a pathological process of abnormal hyperplasia of intrahepatic connective tissues and excessive accumulation of extracellular matrix(ECM)caused by a variety of pathogenic factors,which can ultimately develop into hepatic cirrhosis.Pathogenic factors that cause hepatic fibrosis mainly include chronic hepatitis viruses(HBV and HCV)and parasite(schistosome)infection,alcoholic liver disease,fatty liver disease,cholestasis,drug-induced liver injury,and autoimmune hepatitis.The persistent presence of these factors causes the production and activation of hepatic fibrosis-initiating cells,especially the activation of hepatic stellate cells(HSCs).In addition,there are also fibroblasts,liver sinusoidal endothelial cells,and Kupffer cells surrounding the portal vein,as well as mesenchymal stem cells derived from bone marrow,fibrocytes from blood,and hepatocytes or biliary epithelial cells that have undergone epithelial-mesenchymal transition(EMT).These cells can proliferate,activate,and transdifferentiate into myofibroblasts(MFs)and then synthesize and secret a large amount of ECM,causing hepatic fibrosis and cirrhosis.Mechanistic studies on the development of hepatic fibrosis indicate that many factors associated with liver injuries can induce HSC activation,while HSC activation plays an important role in the development of hepatic fibrosis.The injury of hepatic parenchymal cells and the synthesis and release of a variety of cytokines caused by a variety of pathological factors,including the transforming growth factor ?(TGF-?),epidermal growth factor(EGF),platelet-derived growth factor(PDGF),endothelin(ET),fibroblast growth factor(FGF),connective tissue growth factor(CTGF),and leptin,activate the TGF-?/Smad,epidermal growth factor receptor(EGFR),and Ras/ERK signaling pathways,promote HSC proliferation and transdifferentiation towards MFs.MFs can express a-smooth muscle actin(?-SMA),secrete ECM components including collagens(type I and type?)and proteoglycan to cause ECM accumulation,and promote the development of hepatic fibrosis.Therefore,inhibition of the development of hepatic fibrosis through the inhibition of HSC activation is a major direction in the prevention and treatment of hepatic cirrhosis.There are two main treatment modes of hepatic fibrosis.One is the elimination of pathogenic factors that cause hepatic fibrosis,such as anti-viral therapy and glucocorticoid treatment of autoimmune or alcoholic liver diseases;the other is targeting ECM accumulation to decrease its synthesis and stimulate its degradation.Currently,treatment measures for anti-hepatic fibrosis in the clinic are limited.Therefore,it is necessary to study new types of anti-hepatic fibrosis drugs.Matrine(MT)is the effective component of Sophora flavescens Ait.It has anti-inflammation,immune-suppressive,anti-tumor,and anti-fibrosis activities and can prevent or protect acute liver injury in mice and experimental hepatic fibrosis in rats.However,the pharmacological activity of MT is not strong,its half-life is short,and its functional target is not clear;thus,its anti-fibrosis efficacy in clinical practice is poor.We performed structural modification of MT and obtained a novel thio-derivative of MT,named MD1(C30N4H40SO5F),through thiosulfate and side chain Michael addition to prepare maleate for anti-hepatic fibrosis experiments.First,although matrine(MT)has been reported to have anti-inflammation,immune-suppressive,anti-tumor,and anti-hepatic fibrosis activities,its pharmacological effects still need to be strengthened due to its relatively low efficacy and short half-life,and its underlying mechanisms are largely unknown.For the first time we report a more effective thio derivative of MT,MD-1,and its inhibitory effects on the activation of hepatic stellate cells(HSCs)in both cell culture and animal models.Our study shows that MD-1 can inhibit the proliferation of HSC-T6 cells with the low half-maximal inhibitory concentration(IC50)of 62 ?M.MD-1 also has stronger effects than MT in inhibiting the migration of HSC-T6 cells and can more effectively induce G0/G1 arrest and apoptosis in HSC-T6 cells.Second,our study also investigated the biological mechanisms underlying anti-hepatic fibrosis in the presence of MD-1.We found that MD-1 binds the epidermal growth factor receptor(EGFR)on the surface of HSC-T6 cells,which can further inhibit the phosphorylation of EGFR and its downstream protein kinase B(Akt),resulting in decreased expression of cyclin D1 and eventual inhibition of the activation of HSC-T6 cells.Third,we examined the effect of MD-1 in animal models.In rats with dimethylnitrosamine(DMN)-induced hepatic fibrosis,we found that MD-1 can slow down the development and progression of hepatic fibrosis,protecting hepatic parenchymal cells and improving hepatic functions.In summary,this study investigated the effects of MD-1 on HSC activation and inhibition of hepatic fibrosis in rats.In addition,the possible mechanism of action of this novel MT derivative was investigated to provide potential treatment options for hepatic fibrosis.