Objective To construct lentiviral vectors with different TREM2 truncations and to observe their effects on AML12 hepatocytes under non-alcoholic fatty liver disease(NAFLD)conditions in vitro,laying a theoretical foundation for subsequent macrophage-targeted therapy.Methods Stable transfection of AML12 hepatocyte cell lines with either TREM2 or soluble TREM2(sTREM2)was achieved by lentiviral packaging,and the transfection efficiency was detected using qRT-PCR,Western blotting,and flow cytometry.Immunofluorescence staining was used to detect the expression level and intracellular localization of TREM2 or sTREM2.Flow cytometry was applied to determine the effects of TREM2 or sTREM2 on apoptosis under NAFLD conditions in vitro and on cell cycle under homeostatic conditions.Oil red O staining was utilized to visualize changes in lipid deposition.Results In comparison to the control group,the mRNA and protein expression levels of TREM2 and sTREM2 were significantly increased in the stably transfected cell lines,thus confirming the successful establishment of these cell lines.Notably,under NAFLD conditions in vitro,the overexpression of TREM2 reduced apoptosis and lipid deposition in AML12 hepatocytes(P＜0.05).Under homeostatic conditions,the overexpression of TREM2 or sTREM2 led to a decrease in the proportion of cells in G1 phase(P＜0.05)and an increase in the proportion of cells in S phase(P＜0.01),while the proportion of cells in S phase decreased(P＜0.01)and the proportion of cells in G2 phase increased(P＜0.01)under NAFLD conditions in vitro.Conclusion Overexpression of TREM2 in hepatocytes can reduce hepatocyte apoptosis and lipid deposition,thereby protecting hepatocytes from damage under NAFLD conditions.
维普
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