Dihydroartemisinin Alleviates Steatosis And Inflammation In Nonalcoholic Steatohepatitis By Decreasing Endoplasmic Reticulum Stress And Oxidative Stress

Objective: Non-alcoholic steatohepatitis(NASH)is a severe inflammatory subtype of non-alcoholic steatohepatitis.Metabolic abnormalities promoting hepatic steatosis and inflammation are currently thought to be key to the development of NASH,yet specific therapeutic modalities for the treatment of NASH are lacking.Recent studies have shown that dihydroartemisinin(DHA)has broad pharmacological effects,acting on various organs throughout the body,exerting anti-inflammatory and anti-fibrotic effects.Therefore,we aim to construct a mouse NASH model to clarify the therapeutic effect of DHA on NASH;and to confirm the regulatory mechanism of DHA through ROS regulation of ERS and OS at the molecular and cellular levels.To provide new strategies for the treatment of NASH.Methods: 1.on animal model,WT C57BL/6 mice were used to establish NASH model by western diet pattern of high fat,high cholesterol and high sugar,and the effect of DHA treatment on liver inflammation and lipid accumulation was analyzed by H&E and oil red staining;changes in the levels of ALT,AST and lipid-related indexes(TC,TG,FFA)in serum and tissue homogenate were detected by ELISA and enzymology.WB and IHC analysis of histological levels,changes in the expression of steatosis,inflammation,ERS,OS-related proteins to obtain histological evidence;2.On the cellular model,primary hepatocytes from WT mice were isolated and L02 cell line was used to give cholesterol induction to establish a cellular hyperlipidemic model,and the possible cytotoxicity of DHA to hepatocytes was detected by the proliferation activity of CCK-8 cells,and the possible dangerous dose of DHA was derived possible dangerous dose for treatment;and staining of ROS by immunofluorescence staining and quantification of fluorescence intensity by enzyme marker;extraction of cellular proteins for inflammation of ERS,OS major protein expression.The therapeutic effect of DHA on NASH and the regulatory effect of DHA on ERS and OS were explored by in vivo and in vitro design.Results:We have demonstrated in vitro and in vitro that the therapeutic effect of DHA is effective against NASH,exerting the effect of reducing hepatic steatosis caused by cholesterol and free fatty acid synthesis.It also reduces the expression of inflammatory factors IL-6,IL-1β,TNF-α and inhibits the phosphorylation of NF-κB p65.This is achieved by suppressing excessive unfolded protein response(UPR)and reducing the production of reactive oxygen species(ROS),thereby inhibiting ERS and OS.Conclusion: This study reveals that DHA may reduce hepatic steatosis and inflammation to achieve hepatoprotection and slow down NASH progression.This therapeutic effect was achieved through modulation of ERS and OS exerted to inhibit the release of UPR and ROS,which implies that DHA may be a new and promising candidate for NASH treatment.