锰对多巴胺能神经细胞PC12的毒性及其机制研究

Toxicity effects of manganese on PC12 cells

为探讨锰 (Mn)抑制神经细胞增殖的机制 ,体外培养神经细胞株PC12 ,培养基分别含有 10 0、2 0 0及5 0 0mmol LMnCl2 ,作用 2 4、48、72h后 ,用四唑盐比色实验 (MTT)和平板克隆形成实验检测细胞的存活和生长 ;台盼蓝拒染法绘制生长曲线 ;DTNB法测定谷胱甘肽过氧化物酶 (GSH -Px)的活性、丙二醛比色法测定丙二醛 (MDA)的含量 ;DNA凝胶电泳检测神经细胞凋亡。结果显示 ,MTT和平板克隆形成实验检测结果显示10 0～ 5 0 0mmol L的Mn均对PC12细胞株有显著的抑制作用 ,且呈剂量 效应关系。GSH Px活性降低 ;MDA的活性升高。DNA凝胶电泳结果显示锰能够诱导PC12细胞凋亡。提示锰对神经细胞PC12的增殖具有显著的抑制作用 ,其机制是通过降低过氧化物酶的活性。

In order to study the mechanisms of inhibitory effects of manganese on neurocyte. PC12 cells were incubated in culture media with 100,200 and 500 mmol/L manganese(MnCl 2) for 24, 48 and 72 h respectively. MTT test , Trypan blue exclusion test, malondiadehyde(MDA) colorimetry and DNA gel electrophoresis were performed to exam proliferation, lipid peroxidation, and apoptosis of PC12 cells. The results of MTT test reveled that manganese (100-500 mmol/L) could inhibit the proliferation of PC12 cells. DNA gel electrophoresis showed that the apoptosis of the PC12 cells could be induced by manganese (100 mmol/L). It is concluded that manganese can inhibit proliferation of PC12 cells, the mechanism includes the interference of DNA metabolism, inhibitory of lipid peroxidation, and cell apoptosis.