2,3,5,4'-tetrahydroxy-stilbene-2-O-?-D-glucoside Regulates The Crosstalk Of Macrophages And Hepatocytes In Alcoholic Hepatosteatosis

OBJECTIVE Alcoholic liver disease(ALD)is a major human health problem leading to significant morbidity and mortality in worldwide.The naturally occurring stilbenoid 2,3,5,4'-tetrahydroxy-stilbene-2-O-?-D-glucoside(2354glu)has shown excellent lipid regulatory effects.In this study,we investigated whether 2354glu could inhibit lipid accumulation in liver caused by ethanol and inflammatory factors.METHODS In vivo,mice were treated with ethanol(5g/kg,body weight)every 12 hours and for a total of 3 doses;in the ETOH+2354glu group,mice were tered with ethanol and 2354glu(100mg/kg?50mg/kg);2354glu group were given 2354glu(100 mg/Kg)by intraperitoneal injection.All mice were sacrificed 4 h after the last dosing.The liver protective effect of 2354glu was observed by H&E staining,OIL-RED O staining and immunohistochemistry.The expression of factors related with lipid synthesis and inflammation were observed by western blot,immunohistochemistry,ELISA,QPCR and Immunofluorescence;Inflammation and lipid accumulation in vitro:Mouse peritoneal macrophages were treated with the indicated concentrations of 2354glu for 1 h,and were subsequently activated with LPS for another 4 h followed by 30 min of ATP stimulation,the expression of inflammation related cytokines were detected by western blot and Elisa;HepG2 cells were incubated in normal medium containing LPS/ATP or conditioned medium from LPS/ATP-activated THP-1 macrophages for 24 h.Lipid deposition in HepG2 cells was assessed with Oil red 0 staining,Immunofluorescence and western blot;RESULTS in vivo,H&E staining,OIL-RED 0 staining and immunohistochemical results show that 2354glu can significantly ameliorate liver steatosis caused by alcohol;Immunofluorescence results show that 2354glu significantly reduced P2x7R and NLRP3 inflammasomes positive expression,reduce the macrophages and neutrophils inflammatory recruitment.At the same time,2354glu can increase total and Phosphor-AMPKa and LKB1 protein levels and reduce the expression of SREBP-1 and SREBP-1 target genes;In vitro,P2x7R and inflammation-related cytokines protein expressions were decreased in the presence of 2354glu in mouse peritoneal macrophages;When HepG2 cells were cultured with conditioned medium from LPS/ATP-activated THP-1 macrophages,lipid accumulation in HepG2 cells was greatly decreased by 2354glu;Conclusion:2354glu might ameliorate alcoholic hepatosteatosis mediated by P2x7R-NLRP3 inflammasomes via regulating the crosstalk between activated macrophages and hepatocytes.;...