Based On JAK2/STAT3 Pathway To Study The Mechanism By Which Paeonol Improvement Of Alcoholic Liver Injury

Objective:To explore the mechanism of action of paeonol in ameliorating alcohol-stimulated liver inflammation and oxidative stress injury in mice based on JAK2/STAT3 signaling pathway;and the effect of paeonol on intracellular JAK2/STAT3signaling pathway when LPS stimulated RAW264.7 cells induced inflammatory response.Methods:C57BL/6 mice were randomly divided into normal group,model group,silibinin group(36.8 mg·kg^-1),and tannin low/medium/high(120/240/480 mg·kg^-1)dose group,and the mice in the modeling group were fed with Lieber-De Carli alcohol liquid diet freely,and the drug was administered by gavage from the second day of modeling for 10 days.The mice were observed in the living condition,and the serum biochemical indexes of TG,TC,ALT and AST were measured,and the morphological changes of liver pathology in each group were observed by HE and oil red O staining to verify the reliability of the model.Serum IL-6,IL-1βand TNF-αlevels,liver CAT,GSH,SOD and MDA levels were measured by using kits.Western blot and immunohistochemistry were used to detect the effects of paeonol on the expression of JAK2/STAT3 signaling pathway-related proteins p-JAK2,JAK2,p-STAT3,STAT3 and SOCS3 in mice liver tissues.The expression of SOCS3 protein in liver tissues was detected by immunofluorescence assay.In addition,in vitro experiments were established to investigate the anti-inflammatory effects of tannins based on the JAK2/STAT3 signaling pathway.RAW264.7macrophages were cultured in vitro,and the inflammatory response was induced by LPS,and the cells were divided into blank group,model group,AG490 inhibitor group,and dandruffin administration group for experimental study.The effects of LPS,Pae and AG490 on cell activity were detected by CCK-8,IL-6 and TNF-αcontents in cell supernatant were detected by ELISA kit,p-JAK2,p-STAT3 and SOCS3 protein expression were detected by immunofluorescence,and p-JAK2,JAK2,p-STAT3,STAT3,Western blot SOCS3 protein expression.Results:Compared with the normal group,the serum TG,TC,ALT and AST of each ALD modeling group were increased to different degrees.The results of HE and oil red O staining indicated that there was a large accumulation of lipid droplets in the liver of the ALD modeling group,and the levels of serum inflammatory factors and oxidative stress were increased.Compared with the model group,medium and high doses of dandruff significantly reduced TG,TC,ALT,AST and oxidative stress levels,decreased IL-6,IL-1β,TNF-αexpression,and significantly improved liver pathology in mice with alcohol-induced liver injury.western blot and immunohistochemical and immunofluorescence results showed that the liver tissues of mice in the medium and high doses of dandruff group had p JAK2,p-STAT3 protein expression was decreased and SOCS3 protein expression was increased.Compared with the normal group,after LPS stimulation cells,IL-6 and TNF-αexpression increased,p-JAK2 and p-STAT3 expression were upregulated,and SOCS expression decreased.Pretreatment with tannin down-regulated phosphorylated JAK2and STAT3 expression,which led to an increase in SOCS3 expression.Conclusion:In vitro and in vivo studies showed that dandruffin significantly attenuated liver inflammation and oxidative stress injury in alcoholic liver-injured mice,probably through the regulation of the JAK2/STAT3 signaling pathway.