自噬在米诺环素保护大鼠脑缺血再灌注损伤中的作用研究

doi:10.12092/j.issn.1009-2501.2019.07.002

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (7): 730-736.doi: 10.12092/j.issn.1009-2501.2019.07.002

自噬在米诺环素保护大鼠脑缺血再灌注损伤中的作用研究

肖世庚¹，董文彬²，叶小弟²，陈爱瑛²，程敏²，缪云萍²，郑高利²

¹浙江大学医学院附属第二医院药剂科，杭州 310002，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:2019-01-07 修回日期:2019-05-08 出版日期:2019-07-26 发布日期:2019-07-29
通讯作者: 郑高利，男，博士，研究员，硕士生导师，研究方向：心脑血管药理学。 Tel: 0571-88215620 E-mail: gaoli-z@ 163.com
作者简介:肖世庚，男，硕士，药师，研究方向：心脑血管药理学。 Tel: 15158191075 E-mail: xiaoshigeng0403@zju.edu.cn
基金资助:
国家“重大新药创制”科技重大专项（2013ZX09309005）；浙江省医药卫生平台重点资助计划（2012ZDA009）

Effects of autophagy on the protection of minocycline against cerebral ischemia-reperfusion injury in rats

XIAO Shigeng¹, DONG Wenbin², YE Xiaodi², CHEN Aiying², CHENG Min², MIAO Yunping², ZHENG Gaoli²

¹The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310002, Zhejiang, China; ²Institute of Materia Medica, Zhejiang Academy of Medical sciences, Hangzhou 310013, Zhejiang, China

Received:2019-01-07 Revised:2019-05-08 Online:2019-07-26 Published:2019-07-29

摘要/Abstract

摘要：

目的:探讨自噬在米诺环素保护脑缺血再灌注损伤中的作用。方法:制备大鼠脑中动脉栓塞(MCAO)模型后，注射米诺环素或3-MA进行干预，对大鼠神经功能进行评分；采用TTC法测定脑梗死体积；透射电镜法观察大脑皮层细胞的超微结构及其自噬小体数目的变化；Western blot检测自噬相关蛋白Beclin1、LC3Ⅱ、p62的表达。结果:22.5 mg/kg和45 mg/kg米诺环素组能够显著降低大鼠神经功能评分，减少脑梗死体积，提高LC3Ⅱ和Beclin1的表达，增加p62的降解；高剂量米诺环素（90 mg/kg）组能够进一步提高LC3Ⅱ和Beclin1的表达，然而其大鼠神经功能评分和脑梗死体积却显著增加；自噬小体数也显著增多；当同时给予自噬抑制剂3-MA后，能显著逆转米诺环素（45 mg/kg）的神经保护作用。结论:低剂量米诺环素能够通过增加LC3Ⅱ和Beclin1的表达、促进p62的降解，诱导自噬从而减轻大鼠脑缺血再灌注损伤；而高剂量米诺环素可能是因为其过度激活自噬导致大鼠脑缺血再灌注损伤加重。

关键词: 自噬, 米诺环素, 脑缺血再灌注损伤

Abstract:

AIM: To investigate the effects of autophagy on the protection of minocycline against cerebral ischemia-reperfusion injury in rats. METHODS: Minocycline or Trimethyl adenine was injected to assess the neural function after the middle cerebral artery occlusion (MCAO) model established. The infarct volume was dertemined by TTC staining method at 24h after reperfusion. The ultrastructure of neurone and the quantity changes of autophagic vacuoles were observed by using transmission electron microscopy (TEM). The expression of protein LC3Ⅱ, Beclin1 and p62 were determined by Western blot. RESULTS:Minocycline (22.5 mg/kg, 45 mg/kg) significantly reduced rat nerve function scores and the cerebral infarction volume, while improved the expression of autophagy related proteins LC3Ⅱ and Beclin1 and increased the number of autophagosome and degradation of p62. Minocycline(90 mg/kg) could further increase the expression of LC3Ⅱ and Beclin1, with improvement of neural function and increase of infarct volume. Whereas the neuroprotective effects of minocycline(45 mg/kg) was reversed by the autophagy inhibitor 3-MA. CONCLUSION: Low dose minocycline attenuates cerebral ischemia reperfusion injury in rats, which might be mediated by up-regulating the protein LC3Ⅱand Beclin1 expression and promoting the degradation of p62/SQSTM l and then inducing autophagy; High dose minocycline aggravates cerebral ischemia reperfusion injury in rats by possible over activation of autophagy.

Key words: autophagy, minocycline, cerebral ischemia-reperfusion injury

中图分类号:

R965.2
R54

肖世庚，董文彬，叶小弟，陈爱瑛，程敏，缪云萍，郑高利. 自噬在米诺环素保护大鼠脑缺血再灌注损伤中的作用研究[J]. 中国临床药理学与治疗学, 2019, 24(7): 730-736.

XIAO Shigeng, DONG Wenbin, YE Xiaodi, CHEN Aiying, CHENG Min, MIAO Yunping, ZHENG Gaoli. Effects of autophagy on the protection of minocycline against cerebral ischemia-reperfusion injury in rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(7): 730-736.

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自噬在米诺环素保护大鼠脑缺血再灌注损伤中的作用研究

Effects of autophagy on the protection of minocycline against cerebral ischemia-reperfusion injury in rats

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