Palmitate induces cell apoptosis in insulinoma MIN6 cell / 基础医学与临床

ABSTRACT

Objective To investigate the effect of protein kinase B and its phosphorylation on the apoptosis of MIN6 induced by palmitate.Methods Incubated with different level of palmitate concentration (0 ～ 0.5 mmol/L),MIN6 cells were cultured in high glucose DMEM with or without LY294002 ; the apoptosis of MIN6 cells was detec-ted and quantified with TUNEL technology.Then we inspected the cell ultrastructure through electron microscopy and determined the expression of protein kinase B and its phosphorylation p-PKB (Ser473) by Western blot,fol-lowed by an RT-PCR detection of BAX and BCL-2 expression.Results Apoptosis of MIN6 cells was induced by palmitate in a concentration-dependent correlation and enhanced by LY294002.Palmitate also inhibited phospho-rylation of protein kinase B on Ser473.Finally,we detected a downregulation of BCL-2 mRNA expression and up-regulation of BAX mRNA expression under palmitate-incubated state.Conclusion Palmitate may induce apoptosis of pancreatic β-cells through inhibiting activation of PKB phosphorylation in diabetes.