MnCl_2引起SH-SY5Y细胞内质网应激及Sigma-1受体的保护作用  

Protection of sigma-1 receptor in the endoplasmic reticulum stress induced by MnCl_2 in the SH-SY5Y cells

作　　者：米岚[1] 李振宁[1] 赵鹏[1] 王婷[1] 傅娟玲[1] 姚碧云[1] 周宗灿[1] 

机构地区：[1]北京大学公共卫生学院食品安全毒理学研究与评价北京市重点实验室,北京100191

出　　处：《毒理学杂志》2013年第1期38-42,43,共6页Journal of Toxicology

基　　金：国家自然科学基金项目(30972502)

摘　　要：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,进一步探讨Sigma-1R在锰致神经系统损害中的作用。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0.5、1、2、4、8和24 h处MnCl2对细胞存活的影响,流式细胞技术(FCM)检测上述时间段MnCl2对细胞凋亡的影响,Western blot检测MnCl2作用上述时间后,细胞ERS分子伴侣GRP78、Sigma-1受体(Sigma-1R)、CHOP及凋亡相关蛋白Caspase-4表达的变化,Sigma-1R拮抗剂BD1047抑制其活性4和24 h后,检测细胞凋亡的情况。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GRP78、Sigma-1R、CHOP及Caspase-4的表达;加入拮抗剂BD1047后,在4和24 h细胞存活率明显下降。结论 MnCl2可诱导SH-SY5Y细胞发生ERS,ERS通过上调Sigma-1R的表达,增加细胞的存活率,这可能是预防锰神经毒性的机制之一。Objective To explore whether endoplasmic reticulum stress（ERS） and ERS-mediated apoptosis are involved in the mechanisms of manganese-induced neurotoxicity,further exploring the role of Sigma-1R in the 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;After Sigma-1R agonist BD1047 inhibit its activity for 4 and 24 hours,apoptosis of SH-SY5Y cells was detected by FCM.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.BD1047 can inhibit the activity of Sigma-1R,and MnCl2 can promote the cell apoptosis largely.Conclusion MnCl2 could cause endoplasmic reticulum stress in SH-SY5Y cells and reduce apoptosis via the pathway of ERS by increasing the expression of Sigma-1R.So,Sigma-1R is likely to be involved in protection of the manganese-induced neurotoxicity.

关 键 词：锰 内质网应激 Sigma-1R 保护 SH-SY5Y细胞 

分 类 号：R994.6[医药卫生—毒理学]