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.
出处
《毒理学杂志》
CAS
CSCD
北大核心
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)
