艾灸对类风湿性关节炎大鼠足趾滑膜组织中AMPK/ULK1信号通路的影响

[Effect of moxibustion on AMPK/ULK1 signaling pathway in synovium tissue of toes in rats with rheumatoid arthritis].

作者信息

Liu Li, Zhang Liang, Li Xin, Xu Mei-Li, Wang Wen-Yi, Gong Zhi-Xian, Ai Kun

机构信息

The First Affiliated Hospital/The First Clinical College of Hunan University of Chinese Medicine, Changsha 410007, China.

College of Acupuncture and Massage, Hunan University of Chinese Medicine, Changsha 410208.

出版信息

Zhen Ci Yan Jiu. 2022 Nov 25;47(11):1019-24. doi: 10.13702/j.1000-0607.20220340.

DOI:10.13702/j.1000-0607.20220340

PMID:36453680

Abstract

OBJECTIVE

To observe the effect of moxibustion on AMP-activated protein kinase (AMPK)/UNC-51-like kinase 1 (ULK1) signaling pathway in the synovial tissue of toes in rheumatoid arthritis rats, so as to explore the mechanism of mo-xibustion in the treatment of rheumatoid arthritis (RA).

METHODS

A total of 45 SD rats were randomly divided into blank, model， moxibustion, methotrexate and rapamycin groups, with 9 rats in each group. RA rat model was established by injection of Freund's complete adjuvant. Moxibustion was applied to "Zusanli" (ST36) and "Guanyuan" (CV4) for 20 min, once a day for 3 weeks. Methotrexate group was given methotrexate (0.35 mg/kg) by gavage, twice a week for 3 weeks. Rapamycin group was intraperitoneally injected with rapamycin (1 mg/kg)，once every other day for 3 weeks. The toe volume of the left hind limb of rats was measured by the toe volume measuring instrument. The content of AMP in toe synovium was detected by ELISA. The expression of AMPK and VPS34 protein in toe synovium was detected by Western blot.The expression of ULK1 and Atg13 mRNA in toe synovium was detected by RT-PCR.

RESULTS

Compared with the blank group, the volume of toe in the model group was increased (<0.01)，while the content of AMP, the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly decreased(<0.01).Compared with the model group, the volume of toe in the moxibustion，methotre-xate and rapamycin groups was decreased (<0.05); the content of AMP, the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly increased(<0.05) in the moxibustion group; the content of AMP, the expression of VPS34 protein, the expression of Atg13 mRNA were significantly increased(<0.05) in the methotrexate group; the expression of AMPK and VPS34 proteins, the expression of ULK1 and Atg13 mRNAs were significantly increased (<0.05, <0.01) in the rapamycin group. Compared with the moxibustion group, the expression of AMPK protein in the methotrexate group and the content of AMP in the rapamycin group were significantly decreased (<0.05).

CONCLUSION

Moxibustion can improve joint swelling in RA rats, and the mechanism may be related to promoting the activity of AMPK/ULK1 signaling pathway.

摘要

目的

观察艾灸对类风湿关节炎大鼠足趾滑膜组织中腺苷酸活化蛋白激酶（AMPK）/UNC-51样激酶1（ULK1）信号通路的影响，以探讨艾灸治疗类风湿关节炎（RA）的作用机制。

方法

将45只SD大鼠随机分为空白组、模型组、艾灸组、甲氨蝶呤组和雷帕霉素组，每组9只。采用注射弗氏完全佐剂的方法建立RA大鼠模型。艾灸“足三里”（ST36）和“关元”（CV4）20分钟，每天1次，共3周。甲氨蝶呤组给予甲氨蝶呤（0.35mg/kg）灌胃，每周2次，共3周。雷帕霉素组腹腔注射雷帕霉素（1mg/kg），隔天1次，共3周。用足趾容积测量仪测量大鼠左后肢足趾容积。采用ELISA法检测足趾滑膜中AMP含量。采用Western blot法检测足趾滑膜中AMPK和VPS34蛋白表达。采用RT-PCR法检测足趾滑膜中ULK1和Atg13 mRNA表达。

结果

与空白组比较，模型组足趾容积增大（P<0.01），而AMP含量、AMPK和VPS34蛋白表达、ULK1和Atg13 mRNA表达均明显降低（P<0.01）。与模型组比较，艾灸组、甲氨蝶呤组和雷帕霉素组足趾容积减小（P<0.05）；艾灸组AMP含量、AMPK和VPS34蛋白表达、ULK1和Atg13 mRNA表达均明显升高（P<0.05）；甲氨蝶呤组AMP含量、VPS34蛋白表达、Atg13 mRNA表达均明显升高（P<0.05）；雷帕霉素组AMPK和VPS34蛋白表达、ULK1和Atg13 mRNA表达均明显升高（P<0.05，P<0.01）。与艾灸组比较，甲氨蝶呤组AMPK蛋白表达及雷帕霉素组AMP含量明显降低（P<0.05）。