Study Of Arbutin Effect On The Treatment Of Liver Fibrosis And Its Mechanism

AIM:Fibrosis occurs in many common organs and tissues,it is a process of dynamic damage repair.The damaged area is stimulated and activated by some profibrotic factors and pro-inflammatory factors,and synthetically secretes alpha-smooth muscle actin??-SMA?to the extracellular matrix?ECM?to form fibrosis.Although the pathogenic factors of different fibrotic diseases vary,their common pathological features are the activation of hepatic stellate cells and the excessive deposition of ECM in tissues.Arbutin,also known as arbutin,is a glucose derivative,Studies have shown that arbutin has a good anti-inflammatory effect.This paper aims to study the pharmacodynamic effects of arbutin on liver fibrosis and explore its mechanism.Methods:1.At the animal level,we selected SPF grade C57 mice?8weeks?and randomly divided into three groups:control group,model group and arbutin administration group.The dosage of arbutin is 50 mg/kg.we chose to establish two classic models of biliary ligation and carbon tetrachloride to explore the pharmacodynamic effects and the mechanism of arbutin in the treatment of liver fibrosis.BDL was modeled by bilateral biliary ligation for 15 days,and CCl₄ was injected intraperitoneally for 6weeks?CCl₄:Oil=1:4?.Give arbutin drug treatment while modeling.2.Sample material and biochemical index analysis:After the modeling,the mice were tested for serum sampling and tissue liver extraction.Preliminary activity analysis was performed by related liver function serum index:Alanine aminotransferase?ALT?.3.Liver tissue is mainly divided into two parts,some of which are used for morphological analysis:the degree of fibrosis and efficacy are identified by observation and analysis of pathological experiments.Another part of the liver tissue is used in molecular experiments such as extracting protein?WB?and mRNA to detect changes in the molecular level to determine the molecular mechanism of arbutin in the treatment of liver fibrosis.4.At the cellular level,human hepatic stellate cell line?LX-2?was cultured in a suitable environment to give different concentrations of arbutinto detect the mechanism of arbutin.Result:1.The results of biliary ligation model?BDL?and carbon tetrachloride?CCl₄?biochemical serum showed that the ALT levels of the two models were significantly higher than those of the control group.This data confirmed the successful establishment of the fibrosis model.The arbutin administration group significantly reduced ALT levels and was statistically significant.In the BDL-induced fibrosis model,the arbutin-administered group showed a significant down-regulation trend compared to the two serum markers TBIL and AKP of the model group and was statistically significant.This result indicates that arbutin can effectively alleviate liver injury induced by two different liver fibrosis models.2.The results of pathological experiments showed that compared with the control group,pathologically stained slices in HE and Sirius Red were observed under the microscope.There was a large amount of inflammatory infiltration and necrosis of some hepatocytes in the model group.A large amount of collagen deposition occurred around the loss of normal hepatic lobular structure,and the number of cells of inflammation and necrosis was significantly reduced in the arbutin administration group compared with the model group,and the bridging between collagen fibers in the portal area was improved.3.The results of immunohistochemistry showed that the activation of Marker?alpha smooth muscle actin,?-SMA?in HSCs was significantly reduced in the drug-administered group,and the protein expression of?-SMA in hepatic stellate cells was detected by Western Blot.Fruit glycosides significantly inhibit liver fibrosis.4.The macrophage marker F4/80 was detected by immunofluorescence and real-time quantitative PCR,and it was found that the expression of F4/80 in the arbutin administration group was significantly reduced compared with the model group.This result suggests that arbutin may improve liver fibrosis and may be associated with anti-inflammatory.Subsequently,QPCR was used to detect inflammation-related markers such as tissue inhibitors of matrix metalloproteinase-1?TIMP-1?.Changes in RNA levels demonstrate that arbutin inhibits liver fibrosis by reducing inflammation,and further explores its mechanism of action.5.At the cellular level,we found that the expression of?-SMA in HSCs and p-Smad2,p-Smad3 and p-Akt were significantly down-regulated after adding different concentrations of arbutin in LX-2 cells.The results suggest that arbutin can inhibit liver fibrosis by participating in the p-Akt/Akt and Smad2/3 signaling pathways.CONCLUSION:Arbutin can improve the infiltration of inflammatory cells and the activation of hepatic stellate cells,inhibit collagen production,and thus reduce liver fibrosis.