砷染毒对神经细胞凋亡及calpain 1，calpain 2和cdk5／p25基因和蛋白表达的影响  

Effects of arsenic poisoning on neuronal cell apoptosis and mRNA and protein expression of calpain I, calpain 2, and cdk5/p25

作　　者：李新[1] 常利军[2] 张华俊[2] 张红梅[2] 牛侨[3] 

机构地区：[1]太钢疾病预防控制中心放射卫生科,太原030003 [2]山西医科大学公共卫生学院环境卫生学教研室 [3]公共卫生学院劳动卫生学教研室

出　　处：《中华劳动卫生职业病杂志》2014年第3期202-206,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金项目（81001258）;教育部博士点新教师基金（20091417120003）;山西省优秀青年学术带头人支持计划（晋教财{2011]205号）;山西省青年科技研究基金（2013021033-3）;山西医科大学青年基金（02200919）

摘　　要：目的探讨砷对神经细胞凋亡、calpain1和calpain2以及cdk5／p25基因和蛋白表达的影响，为As2O3神经毒性机制研究提供科学依据。方法原代培养的大鼠神经元，分为未处理对照组、DMSO溶剂对照组，1、5、10Ixmol／LAsA组，分别用0、1、5、10μmol／LAs2O3，溶液染毒，未处理对照组只加人培养液。染毒8h后，用流式细胞术检测神经细胞凋亡率，实时荧光定量PCR法检测calpain1、ca｜pain2、cdk5、p35基因的表达，用蛋白免疫印迹法（Westernbot）检测calpainl、calpain2、cdk5和p35以及p25的蛋白表达水平。结果与未处理对照组和溶剂对照组比较，5和10μmol／LAs2O3染毒组细胞凋亡率明显增加，差异有统计学意义（P〈0．05）。1、5和10μmol／LAs2O3染毒组calpain1基因表达量分别为（6．36±3．26），（7．11±5．13）和（7．47±2．59），cdk5基因表达量分别为（1．27±0．19）、（1．54±0．04）和（1．79±0．21），明显高于未处理对照组[（0．72±0．15）、（1．77±0．87）]和溶剂对照组（0．96±1．23）、（1．18±0．09）]，差异有统计学意义（P〈O．05）；1和5μmol／LAs2O3染毒组p35的基因表达量[（2．17±0．59）、（2．51±0．51）]、明显高于未处理对照组（1．26±0．37），差异有统计学意义（P〈0．05）；1、5和10Ixmol／LAs20，染毒组calpain1蛋白表达量分别为（0．37±0．10）、（0．42±0．13）和（0．51±0．18），明显高于未处理对照组（0．11±0．08）和溶剂对照组（0．13±0．07），差异均有统计学意义（P〈0．05）。5和10μmol／LAs2O3染毒组cdk5蛋白表达量分别为（0．34±0．12）和（O．37±0．21），p25蛋白表达量分别为（0．31±0．23）和（0．55±0．16），明显高于未处理对照组和溶剂对照组，差异均有统计学意义（P〈0．05）；5和10μmol／LAs2O3，染毒组p35的蛋白表达量分别为（0.31±0．23）和（0．26±0．16），明显�Objective To study the effect of arsenic on neuronal cell apoptosis and the mRNA and protein expression of calpain 1, ealpain 2, and cyclin-dependent kinases 5 （cdk5）/p25 and to provide a scientific basis for the research on neurotoxie mechanism of amenic trioxide （As203）. Methods Primary cultured rat neurons were divided into untreated eontrol group, dimethyl sulfoxide （DMSO） solvent control group, and 1, 5, and 10 μmol/L As2O3 treated groups. Eight hours after being treated with As203, cell apoptosis rate was determined by flow eytometry, the mRNA expression of ealpain 1, calpain 2, cdk5, and p35 was measured by real-time fluorescence quantitative PCR, and the protein expression of calpain 1, calpain 2, edk5, p35, and p25 was measured by Western blot. Results Compared with those in the untreated control group and DMSO solvent control group, the cell apoptosis rates in the 5 and 10 μmol/L As203 treated groups were significantly increased （P〈0.05）. The mRNA expression levels of calpain 1 were 6.36-±3.26, 7.11,±5.13, and 7.47-±2.59 in the 1, 5, and 10 μmol/L As203 treated groups, respectively, and the mRNA expression levels of cdk5 were 1.27±0.19, 1.54± 0.04, and 1.79,±0.21 in the 1, 5, and 10μmol/L As203 treated groups, respectively, which were significantly higher than those in the untreated group （0.72±0.15 and 1.77±0.87） and those in the DMSO solvent control group （0.96±1.23 and 1.18±0.09） （P〈0.05）. The mRNA expression levels of p35 in the 1 and 5 μmol/L As203 treated groups were 2.17±0.59 and 2.51 ±0.51, respectively, which were significantly higher than that in the untreated control group （1.26±0.37） （P〈0.05）. The protein expression levels of calpain 1 were 0.37i-0.10, 0.42±0.13, and 0.51±0.18 in the 1, 5, and 10 μmol/L As203 treated groups, respectively, which were significantly higher than those in the untreated control group （0.11±0.08） and DMSO solvent control group （0.13±0.07） （P〈0.05）. In the 5 and 10 Ixmol/L As203 treated g

关 键 词：砷 钙激活蛋白酶 基因表达 

分 类 号：R114[医药卫生—卫生毒理学]