MST过表达及亚砷酸钠对SH-SY5Y细胞的影响

Effects of MST Overexpressin and Sodium Arsenite on SH-SY5Y Cells

目的:探讨亚砷酸钠(NaAsO 2)、巯基丙酮酸硫转移酶(MST)基因过表达对SH-SY5Y细胞的作用及机制。方法:将空载体转染细胞、MST过表达细胞各分为对照组、NaAsO 2组、TUDCA(内质网应激阻断剂)组、NaAsO 2联合TUDCA组,采用MTT、Western blot和酶活性检测等方法观察细胞活力、Bax、Bcl-2凋亡相关蛋白表达及Caspase-3活性。结果:50μmoL/L NaAsO 2暴露24 h显著降低空转组细胞活力和Bcl-2蛋白水平(P<0.01),增加Bax蛋白表达及Caspase-3活性(P<0.01);TUDCA预处理可以逆转以上变化,Bcl-2的蛋白表达则在MST基因过表达细胞砷暴露后发生显著增加,但可被TUDCA所拮抗(P<0.01)。结论:内质网应激可能参与砷、内源性H 2S对SH-SY5Y细胞损伤的影响。

Objective:To investigate the effect and mechanism of sodium arsenite(NaAsO 2)and mercaptopyruvate sulfur transferase(MST)overexpression on SH-SY5Y cells.Methods:SH-SY5Y cells transfected with the empty vector or vector carrying MST gene were divided into control group,NaAsO 2 group,TUDCA(endoplasmic reticulum stress blocker)group and NaAsO 2 combined with TUDCA group.MTT was used to measure cell viability.Western blot was used to detect protein expression levels of apoptosis-related Bax,Bcl-2.Caspase-3 activity was measured by enzyme activity assay.Results:In the empty vector group,exposure to 50μmoL/L NaAsO 2 for 24 h significantly reduced cell viability and Bcl-2 protein levels(P<0.01),and increased Bax protein expression and Caspase-3 activity(P<0.01).TUDCA pretreatment could reverse the above changes.MST overexpression significantly increased Bcl-2 protein expression after arsenic exposure,but it was antagonized by TUDCA(P<0.01).Conclusion:Endoplasmic reticulum stress may be involved in arsenic and endogenous H 2S-mediated SH-SY5Y cell damage.