Autophagy activation prevents sevoflurane-induced neurotoxicity in H4 human neuroglioma cells
Autophagy activation prevents sevoflurane-induced neurotoxicity in H4 human neuroglioma cells
复制标题
自噬激活可防止七氟醚诱导的 H4 人神经胶质瘤细胞神经毒性
DOI：
10.1038/aps.2016.6
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发表时间：
2016-05-01
影响因子：
8.2
通讯作者：
Chen, Gang
中科院分区：
文献类型：
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作者：
Zhou, You-fa;Wang, Qing-xia;Chen, Gang
目的：吸入麻醉剂七氟烷可能会引起动物和人类的认知障碍。先前的研究表明，七氟醚会引发内质网应激，并可能导致大鼠海马神经元凋亡。在本研究中，我们检测了七氟醚是否引起自噬及其对七氟醚诱导的神经细胞损伤的贡献。方法：将H4人神经胶质瘤细胞暴露于4.1%七氟醚6小时。分别使用 CCK8 试剂盒和流式细胞术评估细胞活力和凋亡率。使用GFP-LC3质粒转染或透射电子显微镜检测细胞中的自噬体。 Western blotting检测LC3B、p62/SQSTM、C/EBP同源蛋白（CHOP）和葡萄糖相关蛋白78（GRP78）的表达。结果：七氟烷处理可诱导H4细胞凋亡，并显着增加LC3-II水平和GFP-LC3斑点数量，降低p62表达。雷帕霉素（1 μmol/L）激活自噬显着减少七氟烷诱导的细胞凋亡并增加细胞活力，而3-MA（5 mmol/L）抑制自噬则产生相反的效果。此外，七氟烷治疗显着增加了 ER 应激的两种标志蛋白 CHOP 和 GRP78 的表达。 4-苯基丁酸（500 μmol/L）抑制内质网应激，消除了七氟烷诱导的自噬和细胞凋亡，并提高了活力。此外，七氟醚刺激的CHOP和GRP78的表达被雷帕霉素抑制，但被3-MA进一步增强。结论：七氟醚处理诱导内质网应激并激活自噬，从而拮抗七氟醚诱导的H4人神经胶质瘤细胞的凋亡。结果表明，自噬可能是预防七氟烷引起的神经毒性的潜在治疗靶点。
Aim:The inhaled anesthetic sevoflurane may induce cognitive impairment in both animals and humans. Previous study has shown that sevoflurane triggers ER stress and may lead to apoptosis in rat hippocampal neurons. In this study, we examined whether sevoflurane caused autophagy and its contributions to sevoflurane induced neuronal cell injury.Methods:H4 human neuroglioma cells were exposed to 4.1% sevoflurane for 6 h. Cell viability and apoptosis ratio were assessed using a CCK8 kit and flow cytometry, respectively. Autophagosomes in the cells were detected using GFP-LC3 plasmid transfection or transmission electronic microscopy. The expression of LC3B, p62/SQSTM, C/EBP homologous protein (CHOP) and glucose-related protein 78 (GRP78) was assessed with Western blotting.Results:Sevoflurane treatment induced apoptosis and markedly increased the LC3-II level and GFP-LC3 puncta number, decreased p62 expression in H4 cells. Activation of autophagy by rapamycin (1 μmol/L) significantly reduced sevoflurane-induced apoptosis and increased cell viability, whereas inhibition of autophagy with 3-MA (5 mmol/L) caused the opposite effects. Furthermore, sevoflurane treatment markedly increased the expression of CHOP and GRP78, two hallmark proteins of ER stress. Inhibition of ER stress by 4-phenylbutyrate (500 μmol/L) abrogated sevoflurane-induced autophagy and apoptosis, and improved the viability. Moreover, sevoflurane-stimulated expression of CHOP and GRP78 was inhibited by rapamycin, but further enhanced by 3-MA.Conclusion:Sevoflurane treatment induces ER stress and activates autophagy, which antagonizes sevoflurane-induced apoptosis in H4 human neuroglioma cells. The results suggest that autophagy may be a potential therapeutic target in preventing sevoflurane-induced neurotoxicity.