摘要
目的：通过钙离子络合剂BAPTA-AM对抗过氧化氢诱导的神经母细胞瘤细胞氧化应激反应、探讨钙离子对 NLRP3炎症小体介导的细胞变性的影响作用。方法用过氧化氢处理SHSY5Y细胞以制作细胞氧化应激模型，然后用钙离子载体A23187或钙离子络合剂BAPTA-AM处理细胞模型；细胞实验分为4组：单纯过氧化氢处理组、过氧化氢加钙离子载体A23187处理组、过氧化氢加A23187再加BAPTA-AM处理组以及对照组；再分别用Western Blot 分别检测上述4组细胞NLRP3蛋白，用ELISA方法分别检测上述4组细胞的半胱天冬酶（ Caspase-1）和IL-1β蛋白。结果单纯用过氧化氢处理后，细胞中NLRP3表达明显增加（P＜0．05）；加用A23187处理后，细胞蛋白NLRP3继续增加；Caspase-1和IL-1β表达也有显著增加，分别达到（57．1±19．2） pmol／L 和（484．2±49．5）pg／ml，与对照组比较[Caspase-1：（26．8±12．9）pmol／L；IL-1β：（326．9±52．1）pg／ml]差异有统计学意义（P＜0．05， P＜0．01）。再加用高效钙离子络合剂BAPTA-AM阻抑后，细胞蛋白NLRP3、Caspase-1和IL-1β均明显减少。结论钙超载很可能会强化过氧化氢诱导的细胞氧化应激、并产生由NLRP3炎症小体介导的细胞变性作用。
Objective To study calcium chelator BAPTA-AM antagonize the cellular oxidative stress induced by hydrogen peroxide （ H2 O2 ） and to explore the effect of calcium ion on the cell degeneration mediated by NLRP3.Methods The SHSY5Y cell model of oxidative stress was made by hydrogen perox-ide,then the cell model was treated with calcium ion carrier A23187 or BAPTA-AM,a higher efficiency cal-cium chelating agent.The cells were divided into 4 groups：H2O2 treatment group,H2O2＋A23187 group, H2 O2＋A23187 ＋BAPTA-AM group and control group.NLRP3 protein was detected by Western blot,and Caspase-1 and IL-1βwere detected by ELISA.Results NLRP3 expression was significantly increased in cells treated by hydrogen peroxide（P〈0.05） .The NLRP3 protein continued to increase, and the expression of Caspase-1（（57.1±19.2）pmol/L） and IL-1β（（484.2±49.5）pg/ml） protein was also increased signifi-cantly in cells treated by A23187,and the difference had statistically significant for caspase-1 or IL-1βin H2O2＋A23187 group compared with those in control group（Caspase-1：（26.8±12.9）pmol/L,IL-1β：（326.9 ±52.1） pg/ml） （P〈0.05, P〈0.01） .NLRP3,Caspase-1 and IL-1βwere all significantly reduced after adding a high-er efficiency calcium chelating agent BAPTA-AM.Conclusion Calcium overload is likely to enhance the oxidative stress induced by hydrogen peroxide and engender neurodegeneration mediated by NLRP3 inflammasome.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2016年第3期210-214,共5页
Chinese Journal of Behavioral Medicine and Brain Science
基金
贵州省科技厅课题（黔科合LH字[2014]7159）
贵州省卫生计生委课题（gzwkj[2015-1-042]）.
