内质网应激及其介导的凋亡在锰致神经毒性中的作用

Endoplasmic reticulum stress and ERS-mediated apoptosis in manganese-induced neurotoxicity

目的探讨内质网应激(ERS)及其介导的细胞凋亡在锰引起的神经毒性中的作用。方法以人神经母细胞瘤SH-SY5Y细胞为细胞模型,通过MTT比色法检测MnCl2对细胞存活的影响,流式细胞技术(FCM)检测MnCl2对细胞凋亡的影响,Western blot检测MnCl2作用后,细胞ERS分子伴侣BiP、Sigma-1受体(Sig-1R)、CHOP及凋亡相关蛋白Caspase-4表达的变化。结果 MnCl2可呈时间、剂量依赖性地降低细胞存活率,并诱导SH-SY5Y细胞凋亡;MnCl2上调ERS分子伴侣Bip、Sig-1R、CHOP及Caspase-4的表达。结论 MnCl2可诱导SH-SY5Y细胞发生ERS,早期的ERS对细胞具有保护性效应,持续的ERS通过上调CHOP和Cappase 12的表达介导细胞凋亡,这可能是锰神经毒性机制之一。

Objective To explore whether endoplasmic reticulum stress（ERS） and ERS-mediated apoptosis are involved in the mechanisms of manganese-induced neurotoxicity.Methods Using SH-SY5Y cells as the model of dopaminergic neuron,MTT colorimetry test was used to detect the survival state of SH-SY5Y cells in various doses of manganese chloride;apoptosis of SH-SY5Y cells was detected by flow cytometry（FCM）;the expression of endoplasmic reticulum stress chaperone Bip（Grp78）,Sigma-1R,CHOP and Caspase-4 were detected by Western Blot.Results MnCl2 could dose and time-dependently suppress the viability of SH-SY5Y cells and induce SH-SY5Y cells apoptosis.MnCl2 also could promote the expression of ER stress chaperone Bip（Grp78）,Sigma-1R and apoptosis related protein CHOP,Caspase-4.Conclusion MnCl2 could cause endoplasmic reticulum stress in SH-SY5Y cells and induce apoptosis via the pathway of ERS by increasing the expression of CHOP and Caspase4.So,ERS and ERS-mediated apoptosis are involved in the manganese-induced neurotoxicity.