中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (35): 5596-5602.doi: 10.12307/2023.889

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

ChAT/α7nAChR/核因子κB信号途径在类风湿关节炎发病中的免疫调控

李玉彤，刘静淑，李  振   

  1. 山西中医药大学基础医学院，山西省晋中市  030619
  • 收稿日期:2022-11-01 接受日期:2022-12-12 出版日期:2023-12-18 发布日期:2023-06-01
  • 通讯作者: 李振，副教授，硕士生导师，山西中医药大学基础医学院，山西省晋中市 030619
  • 作者简介:李玉彤，女，1997年生，河北省邢台市人，汉族，在读硕士，主要从事风湿免疫性疾病中西医结合研究。
  • 基金资助:
    国家自然科学基金项目(81904034)，项目负责人：李振；山西中医药大学博士启动基金项目(2020BK02)，项目负责人：李振；山西中医药大学优秀青年科学家培育项目(2021PY-QN-04)，项目负责人：李振；基于炎性反应的重大疾病创新药物山西省重点实验室开放基金资助项目(2021sxcxyw01)，项目负责人：李振；山西省中医药管理局科研课题计划(2023ZYYC058)，项目负责人：李振；山西中医药大学研究生创新创业项目(2021CX015)，项目负责人：李玉彤

Immunoregulatory mechanism of choline acetyltransferase/alpha 7 nicotinic acetylcholine receptor/nuclear factor-kappa B signaling pathway in the pathogenesis of rheumatoid arthritis

Li Yutong, Liu Jingshu, Li Zhen   

  1. College of Basic Medical Sciences, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • Received:2022-11-01 Accepted:2022-12-12 Online:2023-12-18 Published:2023-06-01
  • Contact: Li Zhen, Associate professor, Master’s supervisor, College of Basic Medical Sciences, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • About author:Li Yutong, Master candidate, College of Basic Medical Sciences, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81904034 (to LZ); Shanxi University of Chinese Medicine Doctoral Start-up Fund Project, No. 2020BK02 (to LZ); Shanxi University of Chinese Medicine Outstanding Young Scientist Cultivation Project, No. 2021PY-QN-04 (to LZ); Shanxi Province Key Laboratory Open Fund for Inflammatory Response-based Innovative Drugs for Major Diseases, No. 2021sxcxyw01 (to LZ); Scientific Research Plan of Shanxi Provincial Administration of Traditional Chinese Medicine, No. 2023ZYYC058 (to LZ); Postgraduate Innovation and Entrepreneurship Project of Shanxi University of Chinese Medicine, No. 2021CX015 (to LYT)

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摘要：


文题释义：

类风湿关节炎：是一种慢性炎症性自身免疫疾病，表现为骨和软骨的进行性破坏和血管翳的形成，严重者伴脏器损害和机体功能丧失。类风湿关节炎的发病机制尚不完全明晰，涉及免疫细胞的过度活化和成纤维样滑膜细胞的异常增生，与环境、遗传等因素密切相关。
ChAT/α7nAChR/核因子κB信号途径：ChAT/α7nAChR/核因子κB信号途径立足于胆碱能抗炎通路，胆碱乙酰转移酶(ChAT)催化胆碱和乙酰辅酶A在迷走神经远端产生乙酰胆碱，乙酰胆碱活化并结合烟碱型乙酰胆碱7受体(α7nAChR)，抑制核因子κB信号通路的激活，减少炎症因子的产生，从而减轻机体炎症反应。

背景：胆碱能抗炎通路广泛参与类风湿关节炎发生发展，然而胆碱乙酰转移酶(choline acetyltransferase，ChAT)/烟碱型乙酰胆碱7受体(α7 nicotinic acetylcholine receptor，α7nAChR)/核因子κB信号途径在类风湿关节炎发病中的免疫调控机制研究未见报道。
目的：探究类风湿关节炎发病中ChAT/α7nAChR/核因子κB信号途径的免疫调控机制。
方法：取32只雌性Wistar大鼠，采用随机数字表法分为4组，每组8只：除健康对照组外，关节炎模型组、迷走神经切断组和假手术组均采用牛Ⅱ型胶原和弗氏完全/不完全佐剂构建关节炎大鼠模型，造模成功后，迷走神经切断组切断颈部左侧迷走神经，假手术组仅分离迷走神经。术后5周，检测各组大鼠体质量、关节炎评分和关节肿胀度、脾脏及关节病理变化，ELISA检测血清白细胞介素1、白细胞介素6、肿瘤坏死因子α水平，qRT-PCR及Western blot、免疫组化检测ChAT/α7nAChR/核因子κB通路核心基因mRNA及蛋白表达。

结果与结论：①与健康对照组相比，关节炎模型组大鼠体质量下降(P < 0.05)，关节炎评分升高(P < 0.01)，踝关节肿胀度加重，关节间隙减小伴炎细胞浸润，脾脏白髓及生发中心增大增多，血清白细胞介素、白细胞介素6、肿瘤坏死因子α水平升高(P < 0.05，P < 0.01)，关节中α7nAChR、ChAT、IκBα、核因子κB p50/p65的mRNA及蛋白表达升高(P < 0.05，P < 0.01)，在关节面中显著表达；②与关节炎模型组和假手术组相比，迷走神经切断组大鼠体质量下降(P < 0.05)，关节炎评分升高(P < 0.05，P < 0.01)，踝关节肿胀度加重伴畸形，关节间隙消失伴炎细胞浸润，脾脏白髓及生发中心增大融合，血清白细胞介素1、白细胞介素6、肿瘤坏死因子α水平升高(P < 0.05，P < 0.01)，关节中α7nAChR、ChAT、IκBα、核因子κB p50/p65 mRNA及蛋白表达升高(P < 0.05，P < 0.01)，在关节面中显著表达；③结果提示，迷走神经通过调控胆碱能抗炎通路刺激ChAT/α7nAChR/核因子κB信号途径，进而参与类风湿关节炎的疾病进程，提示胆碱能抗炎通路可能是治疗类风湿关节炎的潜在靶标。

https://orcid.org/0000-0003-4465-1086(李玉彤)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: ChAT/α7nAChR/核因子κB, 信号途径, 类风湿关节炎, 胆碱能抗炎通路, 迷走神经, 免疫调控

Abstract: BACKGROUND: The cholinergic anti-inflammatory pathway is widely involved in the development of rheumatoid arthritis. However, the immunoregulatory mechanism of choline acetyltransferase (ChAT)/α7 nicotinic acetylcholine receptor (α7nAChR)/nuclear factor (NF)-κB signaling pathway in the pathogenesis of rheumatoid arthritis has not been reported.
OBJECTIVE: To investigate the immunoregulatory mechanism of ChAT/α7nAChR/NF-κB signaling pathway in the pathogenesis of rheumatoid arthritis. 
METHODS: Thirty-two female Wistar rats were randomly divided into healthy control group, arthritis model group, vagotomy group and sham operation group, with eight rats in each group. Except for the healthy control group, other groups were treated with bovine type-all collagen and Freund’s complete/incomplete adjuvant to construct the rat arthritis model. After successful molding, the vagus nerve was severed from the left vagus nerve in the neck in the vagotomy group, while only the vagus nerve was isolated in the sham operation group. At 5 weeks after surgery, the body mass, arthritis score, joint swelling degree, pathological changes of the spleen and joint were detected. The levels of interleukin-1, interleukin-6 and tumor necrosis factor-α in serum were detected by ELISA. The mRNA and protein expression levels of ChAT/α7nAChR/NF-κB pathway core genes were detected by qRT-PCR and western blot, respectively, and analyzed by immunohistochemistry. 
RESULTS AND CONCLUSION: Compared with the healthy control group, the body mass of rats in the arthritis model group was significantly decreased (P < 0.05), the arthritis score was significantly increased (P < 0.01), the ankle swelling degree was aggravated, the joint space was reduced with inflammatory cell infiltration, and the white pulp and germinal center of the spleen were enlarged and increased. Moreover, the levels of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased, and the mRNA and protein expressions of α7nAChR, ChAT, IκBα, and NF-κBp50/p65 on the joint surface were also significantly increased in the arthritis model group compared with the healthy control group (P < 0.05, P < 0.01). Compared with the arthritis model group and the sham operation group, the body mass of rats in the vagotomy group was significantly decreased (P < 0.05), the arthritis score was significantly increased (P < 0.05, P < 0.01), the swelling degree of the ankle joint was aggravated with deformity, the joint space disappeared with inflammatory cell infiltration, the white pulp and germinal center of the spleen were enlarged and merged. Compared with the arthritis model group and the sham operation group, the levels of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased in the vagotomy group (P < 0.05, P < 0.01), as well as the mRNA and protein expressions of α7nAChR, ChAT, IκBα, and NF-κBp50/p65 were significantly increased on the joint surface (P < 0.05, P < 0.01). To conclude, the vagus nerve regulates the cholinergic anti-inflammatory pathway and stimulates the ChAT/α7nAChR/NF-κB signaling pathway to participate in the disease progression of rheumatoid arthritis. It indicates that the cholinergic anti-inflammatory pathway may be a potential target for the treatment of rheumatoid arthritis.

Key words: ChAT/α7nAChR/NF-κB, signaling pathway, rheumatoid arthritis, cholinergic anti-inflammatory pathway, the vagus nerve, immunoregulation

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