Exploration Of The Therapeutic Effect Of Levo-tetrahydropalmatine On Lipotoxicity In An In Vitro Model Of Non-alcoholic Fatty Liver Disease Based On The AMPK/SIRT1 Pathway

Objective: To investigate the therapeutic effect of levo-tetrahydropalmatine(THP)on lipotoxicity of non-alcoholic fatty liver disease(NAFLD)in vitro model.Methods: The PO combination of oleic acid(OA): palmitic acid(PA)(2:1)was used to induce human hepatoma cells BEL-7402 to establish NAFLD in vitro lipotoxicity model and THP was used for interventional treatment.Cells were divided into control group,model group,and THP group(75 and 100 μmol/L).The intracellular lipid droplets were observed by oil red O staining,the cell proliferation ability was detected by plate cloning experiment,the cell viability was detected by CCK-8 assay,the kit was detected by lactate dehydrogenase(LDH)level,the TC and TG were detected by enzyme labeling,hepatocyte apoptosis was detected by Hoechst 33342 staining.RT-q PCR was used to detect the expression of FASN,IL-1 and IL-6 m RNA.Western Blot technique was used to detect the expression of p-AMPK,AMPK,SIRT1,FAS,HIF-1α,p-JNK,JNK,Bax,Bcl-2,Caspase-3.Results: PO(800:400 μmol/L)induced BEL-7402 cells to establish NAFLD in vitro lipotoxicity model.Compared with the control group,BEL-7402 cells of model group showed significantly increased lipid deposition,apoptosis,LDH,TC and TG,significantly decreased cell viability,significantly increased FASN,IL-1 and IL-6 m RNA expressions,significantly decreased p-AMPK/AMPK,SIRT1 and Bcl-2 proteins expressions,and significantly increased FAS,HIF-1α,p-JNK/JNK,Bax and Caspase-3 expressions(P<0.05～P<0.001).Compared with the model group,the THP groups showed significantly decreased cellullar lipid deposition,apoptosis,LDH,TC and TG,significantly increased cell viability,significantly decreased FASN,IL-1 and IL-6 m RNA expressions,significantly increased pAMPK/AMPK,SIRT1 and Bcl-2 proteins expressions and significantly decreased FAS,HIF-1α,p-JNK/JNK,Bax and Caspase-3 expressions(P<0.05).Conclusion: THP can improve the abnormal lipid metabolism and lipotoxicity of BEL-7402 cells induced by PO.THP may reduce lipid deposition and lipotoxicity by activating AMPK/SIRT1 pathway,and then alleviate hepatocyte apoptosis caused by lipotoxicity.