摘要
目的 研究五价钒致神经细胞凋亡中细胞色素C（Cyt-c）、caspase-9及caspase-3蛋白的变化,为进一步研究五价钒致神经细胞凋亡的作用机制提供资料.方法 用5、10、20 mmol/L的五氧化二钒（V2O5）体外培养大鼠神经元细胞染毒,用原位末端标记法（TUNEL）检测细胞凋亡的程度;蛋白免疫印迹法（Western blot）检测Cyt-c、Caspase-9、Caspase-3蛋白表达.结果 TUNEL检测结果显示,在细胞核中可以见到凋亡小体,随着染毒剂量增高凋亡小体的细胞数逐渐增多.与对照组比较,0、20 mmol/L染毒组凋亡指数明显增高,差异有统计学意义（P＜0.05或P＜0.01）.Western blot法检测结果显示,与对照组比较,5 mmol/L染毒组的Cyt-c、caspase-3蛋白表达明显升高,差异有统计学意义（P＜0.05）; 10、20 mmol/L染毒组的Cyt-c、caspase-9、caspase-3蛋白表达明显升高,差异有统计学意义（P＜0.01）.神经细胞凋亡指数AI与Cyt-c、caspase-9、caspase-3呈正相关（r=0.954,P＜0.01;r=0.938,P＜0.0;r=0.943,P＜0.01）.结论 五价钒能诱导神经细胞凋亡,Cyt-c、caspase-9及caspase-3蛋白表达在五价钒致神经细胞凋亡的过程中发挥了重要的调控作用.
Objective To investigate the roles of cytochrome c （Cyt-c）,caspase-9,and caspase-3 in pentavalent vanadium-induced neuronal apoptosis and to provide a basis for mechanism research.Methods Neurons from rats aged 1-3 days were cultured and treated with vanadium pentoxide （V2O5） at 5,10,or 20 mmol/L.Neuronal apoptosis was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）.The protein levels of Cyt-c,caspase-9,and caspase-3 were determined by Western blot.Results Apoptosis bodies were detected in the nuclei of neurons by TUNEL.The number of neurons with apoptosis bodies increased with increasing dose of V2O5 The apoptosis index （AI） was significantly higher in the 10 and 20 mm/L exposure groups than in the control group （P＜0.05 or P＜0.01）.Western blot showed that the protein expression levels of Cyt-c and caspase-3 significantly increased in the 5 mmol/L exposure group as compared with the control group （P＜0.05）.In the 10 and 20 mmol/L exposure groups,the protein expression of Cyt-c,caspase-9,and caspase-3 all increased as compared with the control group （P＜0.01）.Neuronal AI was positively correlated with Cyt-c,caspase-9,and caspase-3 （r=0.954,P＜0.01; r=0.938,P＜0.01; r=0.943,P＜0.01）.Conclusion Pentavalent vanadium may induce neuronal apoptosis.The protein expression of Cyt-c,caspase-9,and caspase-3 may play an important role in neuronal apoptosis induced by pentavalent vanadium.
出处
《中华劳动卫生职业病杂志》
CAS
CSCD
北大核心
2014年第9期664-667,共4页
Chinese Journal of Industrial Hygiene and Occupational Diseases
