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乙酰胆碱通过作用于α7烟碱型乙酰胆碱受体抑制大鼠小胶质细胞炎性反应

蒋永莹, 李霖, 沈卫星, 邱一华, 彭聿平^*

南通大学医学院生理学系，南通 226001

摘要

小胶质细胞是中枢神经系统中主要的免疫细胞。本研究旨在探讨乙酰胆碱(acetylcholine, ACh)抑制小胶质细胞炎症反应的具体机制。原代培养Sprague-Dawley (SD)大鼠小胶质细胞，脂多糖(lipopolysaccharide, LPS)诱导建立炎症反应模型，ACh处理24 h后，用Western blot检测多种炎性因子、胰岛素样生长因子1 (insulin-like growth factor 1, IGF-1)和α7烟碱型乙酰胆碱受体(α7 nicotinic acetylcholine receptor, α7nAChR)的蛋白表达，用ELISA检测多种炎性因子和IGF-1的释放情况，用慢病毒转染沉默α7nAChR后观察ACh作用的变化。结果显示，LPS可促进小胶质细胞的激活，上调诱生型一氧化氮合酶(inducible nitric oxide synthase, iNOS)蛋白表达，增加白介素1β (interleukin-1β, IL-1β)和肿瘤坏死因子α (tumor necrosis factor α, TNF-α)的表达和释放，减少神经营养因子IGF-1的表达和释放，ACh能逆转LPS的这些作用；LPS可下调小胶质细胞的α7nAChR蛋白表达，ACh逆转该作用；α7nAChR-shRNA慢病毒转染小胶质细胞后，ACh对抗LPS的作用消失。以上结果提示，ACh对抗LPS诱导的小胶质细胞炎症反应的保护作用是通过α7nAChR实现的，这为今后神经炎症疾病的治疗提供了新的治疗靶点。

关键词： 乙酰胆碱; α7烟碱型乙酰胆碱受体; 小胶质细胞; 脂多糖

分类号：R338

Acetylcholine suppresses microglial inflammatory response in rats via acting on microglial α7nAChR

JIANG Yong-Ying, LI Lin, SHEN Wei-Xing, QIU Yi-Hua, PENG Yu-Ping^*

Department of Physiology, Medical School, Nantong University, Nantong 226001, China

Abstract

Microglia are the main immune cells in the central nervous system. In the present study, the mechanism for acetylcholine (ACh) inhibiting microglial inflammatory response was investigated. Primary culture of microglia was isolated from cerebral cortex of Sprague-Dawley (SD) rats. Lipopolysaccharide (LPS) was used to activate the microglia to induce inflammatory response, and then the microglia were treated with ACh for 24 h. Protein expressions of several inflammatory factors, insulin-like growth factor 1 (IGF-1) and α7 nicotinic acetylcholine receptor (α7nAChR) were detected by Western blot. Release of inflammatory factors and IGF-1 into media was detected by ELISA. After α7nAChR gene silence was achieved by lentivirus-transfection of α7nAChR-shRNA, the change of ACh effect was observed. The results showed that LPS induced microglial activation, up-regulated inducible nitric oxide synthase (iNOS) protein expression, increased the expressions and release of IL-1β and TNF-α, and decreased the expression and release of the neurotrophic factor, IGF-1. ACh could reverse these effects of LPS. Meanwhile, LPS reduced the protein expression of α7nAChR on the microglial cells, whereas ACh could reverse the effect. Silencing of α7nAChR gene in microglia abolished the ability of ACh to inhibit LPS-induced inflammatory responses. These results suggest that ACh exerts its protection against LPS-induced microglial inflammation via acting on α7nAChR on microglia, which may provide a novel target for the treatment of neuro-inflammatory diseases.

Key words: Acetylcholine;;;

收稿日期：2017-06-30　　录用日期：2018-01-24

通讯作者：彭聿平　　E-mail: yppeng@ntu.edu.cn

DOI: 10.13294/j.aps.2018.0011

引用本文：

蒋永莹, 李霖, 沈卫星, 邱一华, 彭聿平. 乙酰胆碱通过作用于α7烟碱型乙酰胆碱受体抑制大鼠小胶质细胞炎性反应[J]. 生理学报 2018; 70 (1): 33-39.

JIANG Yong-Ying, LI Lin, SHEN Wei-Xing, QIU Yi-Hua, PENG Yu-Ping. Acetylcholine suppresses microglial inflammatory response in rats via acting on microglial α7nAChR. Acta Physiol Sin 2018; 70 (1): 33-39 (in Chinese with English abstract).