电针对佐剂性关节炎大鼠滑膜组织中mTOR磷酸化水平的影响

龙轶映 张亮 祁芳 唐丽亚 曹佳男 艾坤 刘梨

〔摘要〕 目的 觀察电针对佐剂性关节炎模型大鼠踝关节滑膜组织中哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin， mTOR） 磷酸化的影响，探索电针治疗风湿性关节炎的作用机制。方法 将48只SD大鼠按随机法分为4组：空白组、模型对照组、电针组、甲氨蝶呤组，每组12只。除空白组外，其余3组于大鼠右后足跖关节处皮内注射弗氏完全佐剂（Freund’s complete

adjuvant， FCA）（0.1 mL/只）进行造模，空白组则同法注射生理盐水。造模后第3天，电针组予以电针“足三里”“关元”和“阿是穴”，隔日1次，共16次;甲氨蝶呤组予以甲氨蝶呤（0.1 μg/kg）灌胃给药，每周1次，共4次。造模后第3、24天测量足跖容积。干预结束后第2天使用ELISA法对各组大鼠血清中肿瘤坏死因子-α（tumor necrosis factor-α， TNF-α）表达量进行检测，Western blot法检测大鼠踝关节滑膜组织p-mTORC1、mTORC1蛋白表达水平。结果 相比于空白组，模型对照组大鼠双侧足跖容积、踝关节滑膜组织中p-mTORC1蛋白表达均增多（P<0.05或P<0.01），且血清中TNF-α含量显著增多（P<0.01）;相比于模型对照组，电针组、甲氨蝶呤组大鼠双侧足跖容积、踝关节滑膜组织p-mTORC1蛋白表达均降低（P<0.05或P<0.01），且大鼠血清中TNF-α含量显著降低（P<0.01）。结论 电针可能通过抑制mTORC1蛋白磷酸化，诱导细胞自噬，缓解炎症反应。

〔关键词〕 佐剂性关节炎;电针;mTOR;细胞自噬;足跖容积;肿瘤坏死因子-α

〔中图分类号〕R245       〔文献标志码〕A        〔文章编号〕doi：10.3969/j.issn.1674-070X.2021.12.012

Effect of Electroacupuncture on mTOR Phosphorylation Lever of Synovial Tissues of

Adjuvant-induced Arthritis Rats

LONG Yiying1， ZHANG Liang1， QI Fang1， TANG Liya1， CAO Jia’nan1， AI Kun1*， LIU Li2*

（1. Hunan University of Chinese Medicine， Changsha， Hunan 410208， China;

2. The First Affiliated Hospital of Hunan University of Chinese Medicine， Changsha， Hunan 410007， China）

〔Abstract〕 Objective To research the effects of electroacupuncture on the phosphorylation of mammalian target of rapamycin （mTOR） in the synovial tissue of ankle joint of adjuvant-induced arthritis rats， and explore the mechanism of electroacupuncture in treating  rheumatic arthritis. Methods 48 SD rats were randomly divided into four groups： blank group， model control group， electroacupuncture group， methotrexate group， with 12 rats in each group. Except blank group， the other three groups were intradermal injected with Freund’s complete adjuvant （0.1 mL in each rat） at the foot pad of the right posterior ankle joint for modeling， while the blank group was injected with saline solution in the same way. Three days after the molding， the electroacupuncture group was given electroacupuncture at “Zusanli” （ST36）， “Guanyuan” （BL26） and “Ashi” （experience acupoint） once of everyday， a total of 16 times. Methotrexate group was given methotrexate （0.1 μg/kg） by intragastric administration， once a week for 4 times. Plantar volume was measured before and on day 3 and 24 after modeling. On the second day after intervention， the levels of tumor necrosis factor-α （TNF-α） in serum of rats were measured by ELISA， the protein expression levels of p-mTORC1 and mTORC1 in the synovium tissue were identified by Western blot. Results Compared with blank group， the plantar volume both sides and p-mTORC1 protein expression in synovial tissue of ankle joint were increased （P<0.05， P<0.01）， and the content of TNF-α in serum was significantly increased （P<0.01） in model control group. Compared with model control group， bilateral plantar volume and the expression of p-mTORC1 protein in synovial tissue of ankle joint of rats in electroacupuncture group and methotrexate group were decreased （P<0.05， P<0.01）， and the content of TNF-α in serum was significantly decreased （P<0.01）. Conclusion Electroacupuncture

may induce autophagy and alleviate inflammatory response by inhibiting phosphorylation of mTORC1 protein.

〔Keywords〕 adjuvant-induced arthritis; electricacupuncture; mTOR; autophagy; plantar volume; tumor necrosis factor-α

类风湿性关节炎（rheumatoid arthritis， RA）是一种病因不明的系统性免疫性疾病。临床研究[1-3]表明，电针可以有效减轻关节炎症，减少RA患者晨僵时间、缓解关节疼痛等症状，改善患者日常生活质量。目前认为，电针治疗RA的机制可能与多方面因素密切相关[4]。……