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彭雄,周兰,施雨燕,龚志贤,张亮,艾坤,李鑫,陈青,刘梨.艾灸对佐剂性关节炎大鼠滑膜组织Beclin-1、LC3-Ⅱ表达的影响[J].湖南中医药大学学报,2024,44(1):122-127[点击复制] |
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| 艾灸对佐剂性关节炎大鼠滑膜组织Beclin-1、LC3-Ⅱ表达的影响 |
| 彭雄,周兰,施雨燕,龚志贤,张亮,艾坤,李鑫,陈青,刘梨 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208;湖南中医药大学中医诊断学湖南省重点实验室, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察艾灸对类风湿关节炎（rheumatoid arthritis， RA）大鼠炎症因子白细胞介素-2（interleukin-2，IL-2）及滑膜细胞中自噬相关因子Beclin-1、微管相关蛋白1轻链3-Ⅱ（microtubule-associated protein1 light chain 3-Ⅱ，LC3-Ⅱ）表达的影响，探索艾灸治疗RA的作用机制。方法 将36只雄性SD大鼠随机分为空白组、模型组、甲氨蝶呤组、艾灸组，每组9只。采用弗氏完全佐剂造模法制备RA大鼠模型。造模成功后艾灸组予艾灸足三里、关元，每次20 min，每天1次；甲氨蝶呤组予甲氨蝶呤0.35 mg/kg灌胃，每周2次。空白组、模型组、甲氨蝶呤组每天给予艾灸组大鼠同样时长及强度的捆绑。每组均干预3周。观察大鼠一般情况，采用足趾容积测量仪检测大鼠左后肢足趾容积，ELISA法检测血清中IL-2含量，Western blot检测大鼠踝关节滑膜组织中Beclin-1、LC3-Ⅱ蛋白相对表达量。结果 与模型组比较，甲氨蝶呤组及艾灸组精神一般，反应尚可，体质量恢复，较活跃，摄食、饮水尚可，足部肿胀、红肿缓解，其局部炎症及全身多发性关节炎情况均轻于模型组。与空白组比较，模型组大鼠于造模后第3、10、17、24天足趾容积明显增大（P<0.01），结合一般情况，提示模型制备成功；甲氨蝶呤组、艾灸组足趾容积于第24天增大（P<0.05）。与模型组比较，甲氨蝶呤组造模后第17、24天足趾容积降低（P<0.05，P<0.01），艾灸组第10、17、24天足趾容积明显降低（P<0.01）。干预3周后，与空白组比较，模型组血清中IL-2含量明显升高（P<0.01），滑膜组织Beclin-1、LC3-Ⅱ蛋白表达量明显上升（P<0.01）；与模型组比较，甲氨蝶呤组、艾灸组血清中IL-2含量降低（P<0.05），艾灸组滑膜组织Beclin-1、LC3-Ⅱ蛋白表达量下降（P<0.05）。结论 艾灸能改善RA大鼠关节肿胀，降低IL-2含量，其作用机制可能是通过调节自噬因子Beclin-1、LC3-Ⅱ蛋白表达量有关。 |
| 关键词: 类风湿关节炎 佐剂性关节炎 滑膜组织 艾灸 白细胞介素-2 Beclin-1 微管相关蛋白1轻链3-Ⅱ |
| DOI：10.3969/j.issn.1674-070X.2024.01.018 |
| 投稿时间：2023-09-30 |
| 基金项目:国家自然科学基金项目（81804204，82074565，82274627）；湖南省中医药管理局重点项目（A2023010）。 |
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| Effects of moxibustion on Beclin-1 and LC3-Ⅱ expressions in synovium of rats with adjuvant-induced arthritis |
| PENG Xiong,ZHOU Lan,SHI Yuyan,GONG Zhixian,ZHANG Liang,AI Kun,LI Xin,CHEN Qing,LIU Li |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Hunan Key Laboratory of TCM Diagnostics, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effects of moxibustion on the inflammatory factor interleukin-2 (IL-2) and the expressions of Beclin-1 and microtubule-associated protein 1 light chain 3-Ⅱ (LC3-Ⅱ) in the synovium of rats with rheumatoid arthritis (RA), and to explore the mechanism of action of moxibustion treating RA. Methods A total of 36 SD male rats were randomly divided into blank, model, methotrexate, and moxibustion groups, with nine rats in each group. The rat model of RA was established by complete Freund's adjuvant. After modeling, moxibustion group was treated with moxibustion at "ZuSanli" (ST36) and "Guanyuan" (CV4) for 20 min, once a day. Methotrexate group was given methotrexate intragastrically (0.35 mg/kg) twice a week. The rats in blank, model, and methotrexate groups were bound for the time and intensity as same as those of moxibustion group daily. All groups were intervened for three weeks. The general condition of the rats was observed. The paw volume of the left hind limb of rats was measured by the paw volume measuring instrument, the content of serum IL-2 was determined by ELISA, and the relative expression levels of Beclin-1 and LC3-Ⅱ proteins in the ankle synovial tissues of the rats were determined by Western blot. Results Compared with model group, the spirit of the rats in methotrexate and moxibustion groups was average, their reaction and intake of food and water were passable with more activities, the body weight was restored, the redness and swelling of their paws were alleviated, and the local inflammation and systemic polyarthritis were less severe than those in model group. Compared with blank group, the paw volume of the rats in model group increased on the 3rd, 10th, 17th, and 24th days after modeling (P<0.01), which, along with the general condition of the rats, suggested a successful modeling; meanwhile, the paw volume of the rats in methotrexate and moxibustion groups increased on the 24th day (P<0.05). Compared with model group, the paw volume of the rats in methotrexate group significantly decreased on the 17th and 24th days after modeling (P<0.05, P<0.01), and that in moxibustion group significantly decreased on the 10th, 17th, and 24th days (P<0.01). After three-week intervention, the expression level of serum IL-2 in model group was significantly elevated (P<0.01), and the protein expression levels of Beclin-1 and LC3-Ⅱ in the synovial tissues were significantly higher (P<0.01); compared with model group, the expression level of serum IL-2 in methotrexate and moxibustion groups was reduced significantly (P<0.05), and the protein expression levels of Beclin-1 and LC3-Ⅱ in the synovial tissues of moxibustion group were lower significantly (P<0.05). Conclusion Moxibustion can alleviate the joint swelling of RA rats and decrease the content of IL-2. The mechanism of action may be related to regulating the protein expression levels of Beclin-1 and LC3-Ⅱ. |
| Key words: rheumatoid arthritis adjuvant-induced arthritis synovial tissue moxibustion interleukin-2 Beclin-1 microtubule-associated protein 1 light chain 3-Ⅱ |
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