铅对培养大鼠海马神经元mGluR5 mRNA和蛋白表达水平的影响  

Effects of lead exposure on expression of mGluR5 in mRNA and protein levels of the cultured hippocampal neurons

作　　者：谢寒芳[1] 颜崇淮[1] 徐健[1] 高宇[1] 余晓刚 潘骏[1] 余晓丹[1] 沈晓明[1] 

机构地区：[1]上海交通大学医学院附属新华医院上海市儿科医学研究所上海市环境与儿童健康重点实验室,200092

出　　处：《中华预防医学杂志》2007年第4期266-270,共5页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金(30070665);上海市科委科技启明星计划资助项目(99QB14008)

摘　　要：目的研究铅暴露对海马神经元代谢性谷氨酸受体5亚型(mGluR5)mRNA 和蛋白水平表达的影响。方法对原代培养3 d 的胎鼠海马神经元分为4组,分别为加入氯化铅溶液终浓度为10^(-8)mol/L、10^(-6)mol/L、10^(-4)mol/L 的各铅暴露组和空白对照组。用实时荧光定量 RT-PCR 和Western blot 方法检测每组神经元 mGluR5 mRNA 和蛋白水平的表达。结果铅暴露组海马神经元mGluR5 mRNA 表达较正常对照组下调,且随铅暴露浓度升高下调越明显。10^(-8)mol/L pb^(2+)组、10^(-6)moL/L pb^(2+)组、10^(-4)mol/L pb^(2+)组 mGluR5 mRNA 表达量与正常对照组比值分别为0.724、0.421、0.321。Western blot 结果显示铅暴露组 mGluR5蛋白表达量低于正常对照组,同样呈剂量效应关系,差异有统计学意义(F=77.966,P<0.001)。结论铅暴露导致培养的大鼠海马神经元 mGluR5基因 mRNA 和蛋白表达下降,且铅暴露剂量与下降幅度有一定相关性。Objective To study the effects on the expression of mGluR5 mRNA and protein levels in primarily cultured hippocampal neurons after lead exposure. Methods Primary embryonic rat hippocampal neuronal culture was prepared. On the 3^rd day of incubation, lead chloride solution was added into medium to produce four different lead exposure levels： 0, 1× 10^-8 mol/L, 1 × 10^-6mol/L, 1 × 10^-4 mol/L Pb^2＋. After 10 days of incubation, the neurons were collected to measure the alteration of mGluR5 mRNA expression by real-time fluorescent quantity PCR and the expression of mGluR5 in protein level by Westernblot. Results The studies revealed that mGluR5 mRNA expression was down-regulated after lead exposure in a dose-dependent manner. The mGluP,.5 mRNA expression of the lower lead-exposed neurons （Pb^2＋ 10^-8 mol/L）, the medium lead-exposed neurons（ Pb^2＋ 10^-6 mol/L）, the higher lead-exposed neurons （Pb^2＋ 10^-4 mol/L） were 0. 724, 0. 421, 0. 321 times less than that of the controls, respectively. The Westernblot demonstrated that mGluR5 expression in protein level should be decreased after lead exposure. Conclusion The expression of mGluR5 in mRNA and protein levels should be down-regulated after lead exposure at different lead levels in a dose-dependent manner.

关 键 词：铅 铅中毒 神经系统 海马 受体 谷氨酸 大鼠 Sprague—Dawley 

分 类 号：R114[医药卫生—卫生毒理学]