自噬在米诺环素保护PC12细胞氧糖剥夺-复糖复氧中的作用

中国临床药理学与治疗学 ›› 2015, Vol. 20 ›› Issue (2): 145-150.

自噬在米诺环素保护PC12细胞氧糖剥夺-复糖复氧中的作用

肖世庚¹, 董文彬², 程敏², 叶小弟², 郑高利^1,2

¹浙江中医药大学药学院,杭州 310053,浙江;
²浙江省医学科学院药物研究所,杭州 310013,浙江

收稿日期:2014-09-12 修回日期:2014-12-10 出版日期:2015-02-26 发布日期:2015-03-20
通讯作者: 郑高利,男,博士,研究员,硕士生导师,研究方向为心脑血管药理学及药物安全性评价。Tel: 0571-88215620 E-mail: mgaoli-z@163.com
作者简介:肖世庚,男,硕士研究生,研究方向为心脑血管药理学。Tel: 0571- 88215620 E-mail: xsg.111@163.com
基金资助:
浙江省医药卫生平台重点资助计划(2012ZDA009);国家“重大新药创制”科技重大专项(2013ZX09309005)

Autophagy activation contributes to the protection of minocycline against oxygen-glucose deprivation and reperfusion in PC12 cells

XIAO Shi-geng¹, DONG Wen-bin², CHENG Min², YE Xiao-di², ZHENG Gao-li²

¹College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang, China;
²Institute of Materia Medica, Zhejiang Academy of Medical sciences, Hangzhou 310013, Zhejiang, China

Received:2014-09-12 Revised:2014-12-10 Online:2015-02-26 Published:2015-03-20

摘要/Abstract

摘要： 目的: 探讨自噬在米诺环素保护PC12细胞氧糖剥夺-复糖复氧中的作用。方法: 构建PC12细胞氧糖剥夺-复糖复氧模型,采用米诺环素(MC,1、10、100 μmol/L)或3-甲基腺嘌呤(3-MA,5 mmol/L)进行干预,MTT检测细胞活力,单丹磺酰戊二胺(MDC)染色观察自噬泡,Western blot检测自噬相关蛋白Beclin1、微管相关蛋白轻链3(LC3Ⅱ)、泛素结合蛋白P62/SQSTM l的表达。结果: 氧糖剥夺处理后,1 μmol/L 和 10 μmol/L的MC能显著提高PC12细胞的存活率,而 100 μmol/L 则会加重细胞的损伤。蛋白LC3Ⅱ和Beclin1的表达与MC的浓度呈正相关性,自噬抑制剂3-MA能够降低LC3Ⅱ和Beclin1,增加P62/SQSTM l的表达逆转MC的保护作用。结论: 低剂量(1、10 μmol/L)MC能够通过增加LC3Ⅱ和Beclin1的表达,诱导PC12细胞产生自噬从而保护氧糖剥夺-复糖复氧处理后的PC12细胞。

关键词: 自噬, 米诺环素, PC12细胞, 氧糖剥夺

Abstract: AIM: To investigate the effects of autophagy on the protection of minocycline in PC12 cells after oxygen-glucose deprivation and reperfusion.METHODS: Cultured PC12 cells were exposed to oxygen-glucose deprivation and then reperfusion, and treated with minocycline (1, 10, 100 μmol/L) or trimethyladenine (3-MA,5 mmol/L) during reperfusion. Cell viability were determined by MTT assay after 6 h of reperfusion. We also stained cells with monodansylcadaverin to observe autophagic vacuole. The expression of protein LC3Ⅱ, Beclin1 and P62/SQSTM l were determined by western blot.RESULTS: Low dose (1, 10 μmol/L) minocycline significantly attenuated cell death cascades after oxygen-glucose deprivation, while the high dose (100 μmol/L) exacerbated the cell injury. The expression levels of LC3Ⅱ and Beclin1 increased more pronouncedly when PC12 cells were treated with minocycline at the higher concentration (1-100 μmol/L). The autophagy inhibitor 3-MA reversed the protection of minocycline by reducing the expression of LC3Ⅱ and Beclin1 and increasing P62/SQSTMl.CONCLUSION: The study suggests that minocycline promotes PC12 cells survival after oxygen-glucose deprivation and reperfusion, which might be mediated by up-regulating LC3Ⅱand Beclin1 expression and inducing autophagy.

Key words: autophagy, minocycline, PC12 cells, oxygen-glucose deprivation

中图分类号:

R965.2

肖世庚, 董文彬, 程敏, 叶小弟, 郑高利. 自噬在米诺环素保护PC12细胞氧糖剥夺-复糖复氧中的作用[J]. 中国临床药理学与治疗学, 2015, 20(2): 145-150.

XIAO Shi-geng, DONG Wen-bin, CHENG Min, YE Xiao-di, ZHENG Gao-li. Autophagy activation contributes to the protection of minocycline against oxygen-glucose deprivation and reperfusion in PC12 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(2): 145-150.

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