1,25(OH)2D3对棕榈酸诱导的小鼠AML12细胞脂质堆积和氧化应激的影响及其作用机制OA

Objective To explore the effect of 1,25-dihydroxyvitamin D3,namely 1,25(OH)2D3,on palmitic acid(PA)-induced lipid accumulation and oxidative stress in AML12 cells of mice and its mechanism.Methods Mouse AML12 cells were assigned to control group,or 1 nM,5 nM,10 nM,50 nM,100 nM,200 nM,and 300 nM 1,25(OH)₂D₃groups.The control group was added with DMEM-F12 complete medium only,while the remaining groups were added with DMEM-F12 complete medium containing corresponding concentrations of 1,25(OH)₂D₃ for 24-hour intervention.In addition,some mouse AML12 cells were obtained and treated with 200 µM PA for 24-hour intervention to construct a non-alcoholic fatty liver disease(NAFLD)cell model.The successfully constructed NAFLD cells were divided into PA model group,or PA+1 nM,PA+5 nM,PA+10 nM,PA+50 nM,PA+100 nM,PA+200 nM,and PA+300 nM 1,25(OH)₂D₃groups.The PA model group was treated with DMEM-F12 complete medium only,while the remaining groups were treated with DMEM-F12 complete medium containing corresponding concentrations of 1,25(OH)₂D₃ for 24-hour intervention.The proliferative activity of mouse AML12 cells in each group was then measured to determine the optimal intervention concentration of 1,25(OH)₂D₃(100 nM).The control group,PA model group,PA+100 nM 1,25(OH)₂D₃ group,and 100 nM 1,25(OH)₂D₃ group were selected to measure mouse AML12 cell proliferative activity,intracellular contents of reactive oxygen species,malondialdehyde,and triglyceride(TG),as well as the expressions of silencing information regulator 2 related enzyme 1(Sirt1),adenosine 5'-monophosphate activated protein kinase(AMPKα),and phosphorylated AMPKα(p-AMPKα).Results The proliferative activity of mouse AML12 cells in the PA+1 nM,PA+5 nM,PA+10 nM,PA+50 nM,PA+100 nM,PA+200 nM,and PA+300 nM 1,25(OH)₂D₃ groups was higher than that in the PA model group.Moreover,the proliferative activity of mouse AML12 cells in the PA+100 nM 1,25(OH)₂D₃ group was higher than that in the PA+the remaining concentrations of 1,25(OH)₂D₃ groups(P<0.05).Compared with the control group,the PA model group exhibited decreased proliferative activity of mouse AML12 cell proliferation,increased contents of TG,reactive oxygen species,and malondialdehyde,and decreased expressions of Sirt1,p-AMPKα,and AMPKα(P<0.05).Compared with the PA model group,the PA+100 nM 1,25(OH)₂D₃ group and the 100 nM 1,25(OH)₂D₃ group showed decreased contents of TG,reactive oxygen species,and malondialdehyde,and increased expressions of Sirt1,p-AMPKα,and AMPKα in mouse AML12 cells(P<0.05).Conclusion 1,25(OH)2D3 can increase the proliferation activity of PA-induced mouse AML12 cells,reduce their lipid accumulation,oxidative stress and lipid peroxidation,and its mechanism may be to activate Sirt1/AMPK signaling pathway and increase the phosphorylation of Sirt1 and AMPK,thereby improving the liver injury of NAFLD.