微囊藻毒素-LR对原代大鼠肝细胞p-JNK1／2、p-P44／42、p-P38蛋白水平的影响  

Microcystin-LR Induces Phosphorylation of p-JNK1/2, P44/42, P38 in Primary Cultured Rat Hepatocytes

作　　者：钱小兰[1] 帅怡[2] 王彦琴[2] 肖萍[2] 仲伟鉴[1,2] 

机构地区：[1]复旦大学公共卫生学院营养与食品卫生教研室,公共卫生安全教育部重点实验室,上海200032 [2]上海市疾病预防控制中心毒性评价科,上海200336

出　　处：《环境与职业医学》2013年第5期356-359,364,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(编号:30800935);上海市公共卫生优秀青年人才培养计划(编号:08GWQ004);上海人才发展资金(编号:2011064)

摘　　要：[目的]研究低剂量微囊藻毒素-LR(microcystin-LR,MC-LR)染毒原代大鼠肝细胞对细胞内磷酸化的c-jun氨基末端激酶(p-JNK1/2)、磷酸化的细胞外信号激酶(p-P44/42)、磷酸化的P38丝裂原活化蛋白激酶(p-P38)蛋白水平的影响。[方法]采用改良胶原酶灌流法建立的原代大鼠肝细胞培养模型,以MC-LR终浓度1×10-9、1×10-10、1×10-11、1×10-12mol/L 4个剂量染毒细胞,分别培养至3、6、12、24h 4个时间点后裂解细胞获取总蛋白,用Western法检测分析丝裂原蛋白激酶家族(MAPKs)磷酸化蛋白p-JNK1/2、p-P44/42、p-P38水平。[结果]染毒原代大鼠肝细胞3、6h后p-JNK1/2、p-P44/42、p-P38蛋白表达水平随染毒剂量降低而上升,在1×10-10或1×10-11mol/L剂量组蛋白水平达到峰值后随染毒剂量降低而蛋白水平增幅减小;染毒至12h除p-P38各剂量组蛋白水平升高外,其余目标蛋白各剂量组与对照组相比差异无统计学意义;染毒24h各目标蛋白各剂量组与对照组相比差异无统计学意义。[结论]低剂量微囊藻毒素可促进原代大鼠肝细胞p-JNK1/2、p-P44/42、p-P38水平升高且具有时间效应关系。[ Objective ] To investigate the effect of low-dose mieroeystin-LR （MC-LR） on phosphorylation of JNK1/2, P44/42, and P38 in primary cultured rat hepatoeytes. [ Methods ] Primary rat hepatoeytes were isolated by eollagenase perfusion method. The obtained cells were treated with ! x 10-9, 1 x 10-1~, 1 x 10-ll, 1 x 10-~2 mol/L MC-LR for 3, 6, 12, and 24h, respectively, then harvested and lysed by radio immunoprecipitation assay （RIPA）. The expressions of p-JNK1/2, p-P44/42, and p-P38 were detected by Western blot. [ Rosults ] With the decrease in dose of MC-LR, the expressions of p-JNK1/2, p-P44/42, and p-P38 increased after 3 h and 6h of incubation across all designed concentrations of MC-LR, and a peak was observed at 1 x 10-l~ or 1 x 10-11 mol/L, then the effect intensity decreased with the dose reduction. After the cells exposed for 12h, only p-P38 showed a slight rise, and no significant differences in phosphorylation expression were found between the various dose groups and the control group. No outstanding effect was observed after 24h exposure to MC^LR~ [ Conclusion ] Low-dose of MC-LR could enhance the phosporylation of JNK1/2, P44/42, and P38, and the effect exhibits time-response relationship.

关 键 词：微囊藻毒素-LR 原代大鼠肝细胞 丝裂原活化蛋白激酶 

分 类 号：R114[医药卫生—卫生毒理学]