乙酰胆碱通过调节IL-1β/IL-1RA平衡改善炎症微环境的机制探讨  

Acetylcholine ameliorates inflammatory microenvironment via regulating the balance of IL-1β/IL-1RA

作　　者：马宁[1,2] 纪超 Ning Ma;Chao Ji(Department of Pharmacology,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences&School of Basic Medicine,Peking Union Medical College,Beijing 100005,China;Institute of Pediatrics,the Seventh Medical Center of PLA General Hospital,Beijing 100700,China;National Demonstration Center for Experimental Basic Medical Education(Peking Union Medical College),Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences&School of Basic Medicine,Peking Union Medical College,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所&北京协和医学院基础学院,药理学系,北京100005 [2]中国人民解放军总医院第七医学中心儿科研究所,北京100700 [3]中国医学科学院基础医学研究所&北京协和医学院基础学院,基础医学国家级实验教学示范中心(北京协和医学院),北京100005

出　　处：《Journal of Chinese Pharmaceutical Sciences》2023年第4期260-267,共8页中国药学（英文版）

基　　金：National Natural Science Foundation of China(Grant No.81100801);Peking Union Medical College Small-Scale Characteristic School Education Project。

摘　　要：本研究旨在探讨乙酰胆碱的抗炎机制,为阿尔茨海默病(Alzheimer's disease,AD)炎性病理机制的验证和治疗药物的筛选提供进一步依据。研究首先建立Aβ25−35诱导小胶质细胞系BV-2细胞炎性损伤的体外模型,观察氯化乙酰胆碱(ACh)抑制炎症反应的作用。于有或无α7 nAChR的阻断剂(α-银环蛇毒)存在的情况下,采用ELISA方法评价ACh作用后IL-1β和IL-1RA的表达及二者比率,并分别利用Western blot和EMSA方法测定MAPK(JNK,p38和ERK)及NF-κB通路相关分子的表达及磷酸化水平。结果显示,ACh能够显著降低IL-1β/IL-1RA比率,对抗Aβ25−35诱导的IL-1系统炎症活性变化,维持IL-1亚家族平衡。预先给予α7 nAChR阻断剂能够阻断ACh对IL-1β和IL-1RA蛋白表达的抑制作用。同时,α7 nAChR阻断剂能够阻断ACh对下游ERK1/2 MAPK蛋白的磷酸化水平和NF-κB的活化的调节作用。总之,乙酰胆碱能够抑制Aβ诱导的BV-2细胞中IL-1亚家族的炎症活性、维持该亚家族的平衡稳定,该作用由α7 nAChR介导,有ERK1/2 MAPK和NF-κB通路的参与。该研究为发现探索AD发病的关键机制提供依据,同时也为寻找AD治疗的新靶点提供参考。In the present study,we aimed to explore the anti-inflammatory mechanism of acetylcholine(ACh)and to provide further evidence for the investigation of the inflammatory pathogenesis of Alzheimer’s disease(AD)and its therapeutic drugs.The in vitro model of Aβ25−35-induced inflammatory injury in microglial cell line BV-2 cells was established to observe the anti-inflammatory effect of ACh chloride.In the presence or absence ofα7 nAChR blocker(α-bungarotoxin),the expressions of IL-1βand IL-1RA and their ratio after ACh treatment were evaluated by ELISA.The expression and phosphorylation levels of MAPK(JNK,p38,and ERK1/2)and NF-κB pathway molecules were determined by Western blot analysis or EMSA,respectively.The results showed that ACh could significantly reduce the ratio of IL-1β/IL-1RA,antagonize the inflammatory activity of the IL-1 system induced by Aβ25−35,and restore the viability of BV-2 cells.Pretreatment withα7 nAChR blocker could block the inhibitory effect of ACh on the IL-1β/IL-1RA ratio.Meanwhile,α7 nAChR blocker could block the phosphorylation level of ERK1/2 MAPK protein and NF-κB activation downstream.Our study suggested that ACh could regulate IL-1 system inflammatory response induced by Aβ25−35 in BV-2 cells and maintain IL-1 subfamily balance,which was mediated byα7 nAChR and involved ERK1/2 MAPK and NF-κB pathways.This study provided a basis for exploring AD inflammatory pathogenesis and also a reference for discovering new targets for AD treatment.

关 键 词：乙酰胆碱 IL-1Β IL-1受体拮抗剂 Β-淀粉样蛋白 阿尔茨海默病 

分 类 号：R965[医药卫生—药理学]