中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2023, Vol. 18 ›› Issue (3): 643-651.doi: 10.4103/1673-5374.350211

• 原著:退行性病与再生 • 上一篇    下一篇

跨膜蛋白16F可作为治疗阿尔茨海默病的新靶点

  

  • 出版日期:2023-03-15 发布日期:2022-08-28
  • 基金资助:
    中国国家自然科学基金项目（82072941），辽宁省重点研发计划指导项目（2020JH2/10300044），沈阳市科技计划项目（20-205-4-050）

TMEM16F may be a new therapeutic target for Alzheimer’s disease

Zhi-Qiang Cui1, Xiao-Ying Hu1, Tuo Yang1, Jing-Wei Guan1, Ying Gu1, Hui-Yuan Li1, Hui-Yu Zhang1, Qing-Huan Xiao2, *, Xiao-Hong Sun1, *   

  1. 1Department of Neurology, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning Province, China; 2Department of Ion Channel Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China
  • Online:2023-03-15 Published:2022-08-28
  • Contact: Xiao-Hong Sun, MD, xhsun@cmu.edu.cn; Qing-Huan Xiao, MD, qinghuanxiao12345@163.com.
  • Supported by:
    The present study was supported by the National Natural Science Foundation of China, No. 82072941 (to QHX), Liaoning Province Key R&D Program Guidance Project, No. 2020JH2/10300044 and Science and Technology Plan Project of Shenyang, No. 20-205-4-050 (both to XHS).

PDF

23

摘要：

多种中枢神经系统疾病中可见跨膜蛋白16F的激活， 且参与了小胶质细胞的吞噬作用和转化，小胶质细胞介导的神经炎症是阿尔茨海默病进展的关键因素，但目前较少有研究关注阿尔茨海默病中跨膜蛋白16F对炎症反应的作用。实验首先将小干扰RNA注射到9月龄APP/PS1转基因小鼠双侧海马，建立跨膜蛋白16F敲低阿尔茨海默病动物模型。而后采用Morris水迷宫试验评价其空间记忆能力，并检测小胶质细胞M1/M2表型及NLRP3炎性小体的生物标志物。结果显示：（1）9月龄APP/PS1小鼠脑组织中存在跨膜蛋白16F高表达。敲低跨膜蛋白16F后，模型小鼠空间记忆能力明显改善，小胶质细胞向抗炎M2型转化、NLRP3炎性小体的激活受到抑制，脑组织中细胞凋亡及淀粉样蛋白β沉积显著减少，脑组织损伤明显减轻。（2）以Aβ25-35刺激人小胶质细胞（HMC3）构建阿尔茨海默病小胶质细胞模型，可见细胞中跨膜蛋白16F、诱导型一氧化氮合酶、促炎细胞因子和 NLRP3 炎症小体相关生物标志物NLRP3、ASC和pro-caspase1表达升高；而敲低细胞中跨膜蛋白16F可增强M2表型生物标志物Arg1和Socs3的表达，减少促炎因子白细胞介素1、白细胞介素6和肿瘤坏死因子α的释放，并通过减少下游促炎性因子白细胞介素1β和白细胞介素18的分泌来抑制NLRP3炎性小体的活化。且跨膜蛋白16F 敲低对M1小胶质细胞的作用可被NLRP3激动剂Nigericin部分逆转。（3）由此提示，跨膜蛋白16F可通过参与小胶质细胞极化和NLRP3炎性小体的激活，参与阿尔茨海默病中的神经炎症反应，且实验结果显示抑制跨膜蛋白16F可能是阿尔茨海默病治疗的潜在靶点。

https://orcid.org/0000-0002-8937-6228 (Xiao-Hong Sun)

关键词:

阿尔茨海默病, 神经炎症, 跨膜蛋白16F, 小胶质细胞极化, M1表型, M2表型, 炎性细胞因子, NLRP3炎性小体, 小干扰RNA, 淀粉样蛋白β

Abstract: TMEM16F is involved in many physiological processes such as blood coagulation, cell membrane fusion and bone mineralization. Activation of TMEM16F has been studied in various central nervous system diseases. High TMEM16F level has been also found to participate in microglial phagocytosis and transformation. Microglia-mediated neuroinflammation is a key factor in promoting the progression of Alzheimer’s disease. However, few studies have examined the effects of TMEM16F on neuroinflammation in Alzheimer’s disease. In this study, we established TMEM16F-knockdown AD model in vitro and in vivo to investigate the underlying regulatory mechanism about TMEM16F-mediated neuroinflammation in AD. We performed a Morris water maze test to evaluate the spatial memory ability of animals and detected markers for the microglia M1/M2 phenotype and NLRP3 inflammasome. Our results showed that TMEM16F was elevated in 9-month-old APP/PS1 mice. After TMEM16F knockdown in mice, spatial memory ability was improved, microglia polarization to the M2 phenotype was promoted, NLRP3 inflammasome activation was inhibited, cell apoptosis and Aβ plaque deposition in brain tissue were reduced, and brain injury was alleviated. We used amyloid-beta (Aβ25–35) to stimulate human microglia to construct microglia models of Alzheimer’s disease. The levels of TMEM16F, inducible nitric oxide synthase (iNOS), proinflammatory cytokines and NLRP3 inflammasome-associated biomarkers were higher in Aβ25–35 treated group compared with that in the control group. TMEM16F knockdown enhanced the expression of the M2 phenotype biomarkers Arg1 and Socs3, reduced the release of proinflammatory factors interleukin-1, interleukin-6 and tumor necrosis factor-α, and inhibited NLRP3 inflammasome activation through reducing downstream proinflammatory factors interleukin-1β and interleukin-18. This inhibitory effect of TMEM16F knockdown on M1 microglia was partially reversed by the NLRP3 agonist Nigericin. Our findings suggest that TMEM16F participates in neuroinflammation in Alzheimer’s disease through participating in polarization of microglia and activation of the NLRP3 inflammasome. These results indicate that TMEM16F inhibition may be a potential therapeutic approach for Alzheimer’s disease treatment.

Key words: Alzheimer’s disease, Aβ plaque, inflammatory cytokines, M1 phenotype, M2 phenotype, microglia polarization, neuroinflammation, NLRP3 inflammasome, siRNA, TMEM16F