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 (9): 1813-1820.doi: 10.4103/1673-5374.306094

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

线粒体酸5调控线粒体融合蛋白2保护小胶质细胞存活

  

  • 出版日期:2021-09-15 发布日期:2021-02-05
  • 基金资助:

    湖南省自然科学基金项目（2017JJ3273）

Mitochonic acid 5 regulates mitofusin 2 to protect microglia

Jian Tan, Shuang-Xi Chen#, Qing-Yun Lei, Shan-Qing Yi, Na Wu, Yi-Lin Wang, Zi-Jian Xiao*, Heng Wu*   

  1. Department of Neurology, The First Affiliated Hospital of University of South China, Hengyang, Hunan Province, China
  • Online:2021-09-15 Published:2021-02-05
  • Contact: Heng Wu, PhD, 2915176817@qq.com; Zi-Jian Xiao, PhD, zijian6340@21cn.com.
  • Supported by:
    The study was supported by the Natural Science Foundation of Hunan Province of China, No. 2017JJ3273 (to ZJX).

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摘要：

小胶质细胞凋亡是一种与神经炎症发展相关的重要因素，但是尚无有效方法保护小胶质细胞免受炎症诱导产生细胞凋亡。实验首先将小鼠BV-2细胞（5×106）在10μg/mL脂多糖中培养12 h模拟炎症环境，以5μM线粒体酸5共培养。结果发现5μM线粒体酸5可以显著提高脂多糖干预细胞的存活率，降低与凋亡相关的caspase-3的活性，并减少TUNEL阳性细胞数量，提高细胞中ATP水平，降低mPTP的开放率、钙超载以及Smac的扩散，同时降低凋亡相关蛋白线粒体Smac，细胞质Smac，pro-caspase-3，cleaved-caspase-3和caspase-9的表达，提高抗凋亡蛋白Bcl-2和x-IAP水平以及线粒体相关蛋白线粒体融合蛋白2，线粒体LC3II，自噬相关蛋白Beclin1，p62和Atg5的表达。但是以小干扰RNA沉默线粒体融合蛋白2后，线粒体酸5则不能发挥上述作用。说明线粒体酸5可通过增加与线粒体融合蛋白2相关的线粒体反应，逆转细胞能量产生并维持BV-2细胞对脂多糖诱导的炎症反应的能量代谢。表明线粒体酸5可能通过调控线粒体融合蛋白2促进小胶质细胞的存活。

https://orcid.org/0000-0002-6569-4651 (Heng Wu) 

关键词:

小胶质细胞, 细胞凋亡, 神经炎症, 线粒体酸5, 细胞存活, BV-2细胞, 线粒体融合蛋白2, 线粒体自噬, 能量代谢, 脂多糖

Abstract: Microglial apoptosis is associated with neuroinflammation and no effective strategies are currently available to protect microglia against inflammation-induced apoptosis. Mouse microglial BV-2 cells (5 × 106) were incubated with 10 μg/mL lipopolysaccharides for 12 hours to mimic an inflammatory environment. Then the cells were co-cultured with mitochonic acid 5 (MA-5) for another 12 hours. MA-5 improved the survival of lipopolysaccharide-exposed cells. MA-5 decreased the activity of caspase-3, which is associated with apoptosis. MA-5 reduced the number of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling-positive cells, and increased adenosine triphosphate levels in cells. MA-5 decreased the open state of the mitochondrial permeability transition pore and reduced calcium overload and diffusion of second mitochondria-derived activator of caspase (Smac). MA-5 decreased the expression of apoptosis-related proteins (mitochondrial Smac, cytoplasmic Smac, pro-caspase-3, cleaved-caspase-3, and caspase-9), and increased the levels of anti-apoptotic proteins (Bcl2 and X-linked inhibitor of apoptosis protein), mitochondria-related proteins (mitochondrial fusion protein 2, mitochondrial microtubule-associated proteins 1A/1B light chain 3B II), and autophagy-related proteins (Beclin1, p62 and autophagy related 5). However, MA-5 did not promote mitochondrial homeostasis or decrease microglial apoptosis when Mitofusin 2 expression was silenced. This shows that MA-5 increased Mitofusin 2-related mitophagy, reversed cellular energy production and maintained energy metabolism in BV-2 cells in response to lipopolysaccharide-induced inflammation. These findings indicate that MA-5 may promote the survival of microglial cells via Mitofusin 2-related mitophagy in response to lipopolysaccharide-induced inflammation.

Key words: apoptosis, autophagy related 5, Beclin1, BV-2 cells, caspase-3, inflammation, lipopolysaccharide, mitophagy, Smac, survival