中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (6): 1044-1051.doi: 10.4103/1673-5374.300725

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

Apelin-13抑制脑缺血再灌注损伤后的细胞凋亡和过度自噬

  

  • 出版日期:2021-06-15 发布日期:2020-12-31
  • 基金资助:

    国家自然科学基金项目（81870948，81671276，81501018）；山东省自然科学基金（ZR2014HL040）；济宁医科大学教师研究计划资助基金（JYFC2018KJ003）

Apelin-13 inhibits apoptosis and excessive autophagy in cerebral ischemia/reperfusion injury

Zi-Qi Shao1, Shan-Shan Dou2, Jun-Ge Zhu1, Hui-Qing Wang1, Chun-Mei Wang2, Bao-Hua Cheng2, *, Bo Bai2, *#br#   

  1. 1 Cheeloo College of Medicine, Shandong University, Jinan, Shandong Province, China;  2 Neurobiology Institute, Jining Medical University, Jining, Shandong Province, China
  • Online:2021-06-15 Published:2020-12-31
  • Contact: Bao-Hua Cheng, PhD, chengbh1979@163.com; Bo Bai, PhD, bbai@mail.jnmc.edu.cn.
  • Supported by:
    The study was supported by the National Natural Science Foundation of China, Nos. 81870948 (to BB), 81671276 (to BHC), 81501018 (to CMW); the Natural Science Foundation of Shandong Province of China, No. ZR2014HL040 (to BHC); Program Supporting Foundation for Teachers’ Research of Jining Medical University of China, No. JYFC2018KJ003 (to SSD).

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摘要：

Apelin-13是孤儿G蛋白偶联受体-血管紧张素受体样蛋白J受体的内源性配体，可对脑缺血损伤起到神经保护作用，但作用机制尚待阐明。为探索Apelin-13对脑缺血再灌注损伤模型中细胞凋亡和自噬影响的机制，实验建立了大脑中动脉闭塞诱导的大鼠脑缺血再灌注损伤模型，以右侧脑室注射50 µg/kg Apelin-13进行干预。另外还通过无糖培养基95％N2和5％CO2培养处理4h后，再以含糖培养基培养正常环境培养5h，建立体外氧糖剥夺再灌注诱导的SH-SY5Y细胞模型，并以10-7M Apelin-13干预5h。结果发现，（1）Apelin-13可预防脑缺血再灌注损伤，同时在体内外增加Bcl-2/Bax比值，并降低裂解的caspase-3的表达，抑制神经元的凋亡，还能通过调节LC3B，p62和Beclin1表达抑制过度自噬；同时Bcl-2和PI3K/Akt/mTOR通路蛋白的表达明显增加。（2）利用PI3K/Akt/mTOR通路抑制剂LY294002（20 µM）和雷帕霉素（500 nM）预处理SH-SY5Y细胞，则会显著减弱Apelin-13对自噬和细胞凋亡的抑制作用。（3）实验结果表明Apelin-13可通过上调Bcl-2表达和激活mTOR信号通路，来抑制细胞凋亡和过度自噬，在体内外脑缺血再灌注损伤模型中发挥神经保护作用。实验究于2018年2月经济宁医科大学动物伦理委员会批准，批准号2018-JS-001。

https://orcid.org/0000-0003-1729-8184 (Bao-Hua Cheng); 

https://orcid.org/0000-0001-9988-9318 (Bo Bai)

关键词: 中枢神经系统, 脑, 脑损伤, 因子, 通路, 细胞凋亡, 自噬, 神经保护, 再生

Abstract: Apelin-13 is a novel endogenous ligand for an angiotensin-like orphan G-protein coupled receptor, and it may be neuroprotective against cerebral ischemia injury. However, the precise mechanisms of the effects of apelin-13 remain to be elucidated. To investigate the effects of apelin-13 on apoptosis and autophagy in models of cerebral ischemia/reperfusion injury, a rat model was established by middle cerebral artery occlusion. Apelin-13 (50 µg/kg) was injected into the right ventricle as a treatment. In addition, an SH-SY5Y cell model was established by oxygen-glucose deprivation/reperfusion, with cells first cultured in sugar-free medium with 95% N2 and 5% CO2 for 4 hours and then cultured in a normal environment with sugar-containing medium for 5 hours. This SH-SY5Y cell model was treated with 10–7 M apelin-13 for 5 hours. Results showed that apelin-13 protected against cerebral ischemia/reperfusion injury. Apelin-13 treatment alleviated neuronal apoptosis by increasing the ratio of Bcl-2/Bax and significantly decreasing cleaved caspase-3 expression. In addition, apelin-13 significantly inhibited excessive autophagy by regulating the expression of LC3B, p62, and Beclin1. Furthermore, the expression of Bcl-2 and the phosphatidylinositol-3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway was markedly increased. Both LY294002 
(20 µM) and rapamycin (500 nM), which are inhibitors of the PI3K/Akt/mTOR pathway, significantly attenuated the inhibition of autophagy and apoptosis caused by apelin-13. In conclusion, the findings of the present study suggest that Bcl-2 upregulation and mTOR signaling pathway activation lead to the inhibition of apoptosis and excessive autophagy. These effects are involved in apelin-13-induced neuroprotection against cerebral ischemia/reperfusion injury, both in vivo and in vitro. The study was approved by the Animal Ethical and Welfare Committee of Jining Medical University, China (approval No. 2018-JS-001) in February 2018.  

Key words: central nervous system, brain, brain injury, factor, pathways, apoptosis, autophagy, neuroprotection, regeneration