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目的 以鼠嗜铬神经瘤细胞 (PC12 )为模型 ,筛选锰对神经细胞增殖抑制作用的时间及剂量 ,观察细胞形态学、细胞周期和生化指标改变与丝裂原活化蛋白激酶 (p38MAPKs)活化表达间的关系。方法 用 2 0 0 ,4 0 0 ,6 0 0 ,80 0μmol/LMnCl2 的培养液 ,分别作用对数生长期PC12细胞 1,2 ,3,4d后 ,用噻唑蓝比色 (MTT)筛选锰的细胞毒性剂量 ;流式细胞仪检测细胞周期分布 ;透射电镜观察细胞形态学变化 ;琼脂糖凝胶电泳检测MnCl2 对PC12细胞基因组DNA的影响。蛋白印迹 (western -blot)法检测 p -p38。结果 MTT实验结果显示 ,2 0 0～ 80 0 μmol/LMnCl2 作用 1,2 ,3,4d对PC12有显著的抑制作用 ,呈剂量和时间依赖趋势 ,6 0 0 μmol/LMnCl2 作用 4d对PC12的抑制率可达 5 0 %以上。流式细胞仪检测实验表明 :6 0 0 μmol/LMnCl2 作用 4d将PC12细胞周期阻滞在S期 ,诱导细胞凋亡 ,与电镜结果一致 ,同样条件下细胞DNA碎片化。Western -blot实验显示 6 0 0 μmol/LMnCl2 作用 1,2 ,3,4dp -p38逐渐升高 ,3d时较对照组增加 6 6倍 (n =3,P <0 0 5 ) ,2 0 0 ,4 0 0 ,6 0 0 μmol/LMnCl2 作用 4d时 ,磷酸化蛋白 38(p - p38)也逐渐升高 ,4 0 0 μmol/LMnCl2 作用 4d时较对照组升高 4 7倍 (n =3,P <0 0 5 )。结论 锰通过MEK3/
OBJECTIVE: To determine the time and dose of manganese inhibition on the proliferation of nerve cells by using PC12 as a model, and to observe the changes of cell morphology, cell cycle and biochemical indexes and the activation of p38 MAPKs Relationship between. Methods PC12 cells were treated with 200, 400, 600, 80μmol / L MnCl2 for 1, 2, 3 and 4 days, respectively, and the contents of manganese Cytotoxicity, cell cycle distribution were detected by flow cytometry, cell morphological changes were observed by transmission electron microscopy, and the effect of MnCl2 on genomic DNA of PC12 cells was detected by agarose gel electrophoresis. P -p38 was detected by western-blot. Results The results of MTT assay showed that PC12 was inhibited by 1,200 and 800 μmol / L MnCl2 for 1, 2, 3 and 4 days in a dose- and time-dependent manner. PC12 was inhibited by 600 μmol / L MgCl2 for 4 days Rate up to 50% or more. Flow cytometry test showed that: PC12 cells were blocked by S phase at 600 μmol / L LMnCl 2 for 4 days to induce apoptosis, which was consistent with the electron microscopy. The results of Western-blot showed that the effect of 6 0 0 μmol / L MnCl2 on 1,2,4 and 4dp-p38 increased gradually, compared with the control group 6 and 6 times (n = 3, P <0 05) The phosphorylation of protein 38 (p - p38) also increased gradually after treated with 400 μmol / L LMnCl2 for 4 days. Compared with the control group, the level of phosphorylation protein 38 (p - p38) increased 4: P <0 0 5). Conclusion Manganese through MEK3 /