镉对体外培养大鼠大脑皮质神经元钙稳态的影响  

Effects of cadmium on calcium homeostasis in cerebral cortical neurons of rat cultured in vitro

作　　者：袁燕[1] 江辰阳[1] 孙娅[1] 刘学忠[1] 顾建红[1] 卞建春 刘宗平[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《中国兽医学报》2013年第3期450-453,471,共5页Chinese Journal of Veterinary Science

基　　金：江苏省高校"青蓝工程"中青年学术带头人培养对象资助项目(2006);国家自然科学基金资助项目(31101866);江苏省自然科学基金资助项目(BK2008214)

摘　　要：选用原代培养大鼠大脑皮质神经元为模型,用神经元特异性烯醇化酶(NSE)抗体经免疫组织化学染色技术鉴定神经元。用不同浓度(0、5、10、20μmol/L)醋酸镉染毒大鼠大脑皮质神经元12h,利用流式细胞仪检测细胞内[Ca2+]i,ATPase酶试剂盒测定Na+-K+-ATPase和Ca2+-Mg2+-ATPase活性的变化,荧光定量PCR法测定钙调蛋白(CaM)mRNA转录水平。结果表明,免疫组化染色证实培养细胞呈现NSE阳性染色,证明是神经元。与对照组相比,各镉染毒组细胞内[Ca2+]i显著或极显著升高(P<0.05或P<0.01),Na+-K+-ATPase和Ca2+-Mg2+-ATPase活性显著或极显著降低(P<0.05或P<0.01),20μmol/L组CaM mRNA转录水平极显著降低(P<0.01)。说明镉可能通过影响CaM的转录水平与维持钙稳态相关的酶(Na+-K+-ATPase和Ca2+-Mg2+-ATPase)活性,干扰神经元细胞内钙稳态,进而造成神经元细胞损伤。To investigate the effect of cadmium on calcium homeostasis in cerebral cortical neurons of rat cultured in vitro ,neurons specific enolase（NSE）antibody, recognized specifically NSE anti- gen in cultured cells,was used to identify cerebral cortical neurons of rat by immunocytochemical method. The neurons were exposured to different concentrations（0,5,10,20 μmol/L of cadmium acetate for 12 hours. Then,intracellular[Ca^2＋] was detected by flow cytometry, the activities of Na＋- K＋-ATPase and Ca^2＋-Mg^2＋-ATPase were measured by ATPase test kits and the transcrip tion level of CaM mRNA was detected by real-time fluorescent quantitative PCR. The neurons were affirmed by NSE staining which showed that cultured cells were positive staining by NSE antibody. In comparison with the control group,the results showed that intracellular[Ca^2＋];was increased significantly （P〈0.05 or P〈 0.01） ,the activities of Na＋ K＋-ATPase and Ca^2＋-Mg^2＋- ATPase were decreased significantly（P〈0.05 or P〈0.01）, the transcription level of CaM mR- NA was decreased significantly（P〈0.01） in 20μmol/L group. It was suggested that cadmium could impair neurons by disturbing intracellular Ca^2＋ homeostasis and inhibiting the transcription level of CaM mRNA and activities of Na＋-K＋-ATPase and Ca^2＋-Mg^2＋-ATpase.

关 键 词：镉 大脑皮质神经元 钙稳态 

分 类 号：S859.8[农业科学—临床兽医学]