三甲基氯化锡诱导PC12细胞凋亡的机制  

Study on mechanism of trimethyltin chloride-induced apoptosis in PC12 cells

作　　者：梁艳芳[1] 卿燕[1] 杜清清[1] 范攀[1] 徐以平[1] 徐汉宫[1] 石年[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院卫生毒理学系,环境与健教育部重点实验室,武汉430030

出　　处：《中华劳动卫生职业病杂志》2012年第11期816-819,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

摘　　要：目的在体外条件下，研究三甲基氯化锡（trimethyltinchloride，TMT）对PCI2细胞增殖、细胞凋亡及细胞氧化损伤的作用，并探讨TMT对NF．KB表达的影响。方法（1）不同浓度TMT（O、0．3125、0．6250、1．2500、2．500、5．000、10．000、20．000μmol／L）处理PCI2细胞24、48h，噻唑蓝（MTF）法测定细胞存活率；（2）1．25、2．50、5．00、10．00μmol／LTMT分别染毒PCI2细胞12、24h后，流式细胞仪检测检测细胞凋亡率；（3）1．25、2．50、5．00、10．00μmol／L剂量的TMT染毒PCI2细胞6h后，测定活性氯（ROS）和谷胱甘肽（GSH）含量的变化：（4）1．25、2．50、5．00、10．00μmol／L剂量的TMT染毒PCI2细胞12h后免疫印迹法（Westernblot）检测核蛋白NF．KB的水平。结果与溶剂对照组比较，染毒24h，2．5000、5．0000、10．0000、20．0000μmol／LTMT剂量组细胞存活率明显下降；染毒48h，1．2500、2．5000、5．0000、10．0000、20．0000trmol／L剂量组细胞存活率明显下降，差异均有统计学意义（P〈0．05）。1．25、2．50、5．00、10．00μmo]／LTMT染毒细胞12h，凋亡率分别为15．30％±0．75％、18．90％±0．61％、22．OO％±0．60％、36．50％±0．66％，染毒24h凋亡率分别为28．60％士0．40％、43．54％±2．00％、65．73％±0．71％、74．67％±0．40％，明显高于对照组[12h：（12．80％±1．00％）、24h：（16．83％±0．25％）]，差异均有统计学意义（P〈0．05）。1．25、2．50、5．00、10．00μmol／L剂量组的ROS荧光强度值分别为对照组的1．42、1．71、1．78、1．89倍，差异有统计学意义（P〈0．05）。2．50、5．00、10．00μmol／L组GSH含量分别为（0.17±0．0）、（0．20±0．04）、（0．07±0．03）μmol／μgpro，明显低于对照组（0．30±0．01）μmol／Lpro，差异有统计学意义（P〈0.05）。2．50、5．00、10．00μm01]L剂量组NF—KBp65表达的蛋白条带灰度值Objective To investigate the Jfects of trimethyltin chloride （TMT） on proliferation, apoptosis, oxidative damage, and NF-KB expression in PC12 cells in vitro. Methods PC12 cells were treated with 0, 0.3125, 0.6250, 1.2500, 2.5000, 5.0000, 10.0000, and 20.0000 μmol/L TMT for 24 and 48 h, and MTY assay was used to evaluate cell viability. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00 μmol/L TMT for 12 and 24 h, and flow cytometry was used to measure the apoptotic rates of cells. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00μmol/L TMT for 6 h, and the reactive oxygen species （ROS） and glutathione （GSH） levels were measured. PC12 cells were treated with 1.25, 2.50, 5.00, and 10.00 μmol/L TMT for 12 h, and Western blot was used to measure NF-KB levels. Results Compared with solvent controls, the PC12 cells treated with 2.5000, 5.0000, 10.0000, and 20.0000 μmol/L TMT for 24 h showed significantly decreased cell viability （P〈0.05）; the PC12 cells treated with 1.2500, 2.5000, 5.0000, 10.0000, and 20.0000 μmol/L TMT for 48 h showed significantly decreased cell viability （P〈0.05）. The PC12 cells treated with 1.2500, 2.5000, 5.0000, and 10.0000μmol/L TMT for 12 h had apoptotic rates of 15.30%±0.75%, 18.90%±0.61%, 22.00%± 0.60%, and 36.50%±0.66%, respectively, and the PC12 cells treated with 1.25, 2.50, 5.00, and 10.00 I±mol/L TMT for 24 h had apoptotic rates of 28.6%±0.40%, 43.54%±2.00%, 65.73%±0.71%, and 74.67%±0.40%, respectively, all significantly higher than those of the control group （ 12 h： 12.80%±1.00%, 24h： 16.83%±0.25%） （P〈0.05）. The ROS fluorescence intensities of the PC12 cells treated with 1.25, 2.50, 5.00, and 10.00μmol/L TMT were 1.42, 1.71, 1.78, and 1.89 times that of the control group （P〈O.05）; the PC12 cells treated with 2.50,5.00, and 10.00 μmol/L TMT had GSH levels of 0.17±0.0, 0.20 ±0.04, and 0.07 ±0.03 μmol/txg protein, significantly lower than that of the control group （0.30±0.01 μmol/L protein） （P〈0.05

关 键 词：三甲基氯化锡 NF-KB 谷胱甘肽 活性氧组分 

分 类 号：R114[医药卫生—卫生毒理学]