Chinese Journal of Tissue Engineering Research ›› 2022, Vol. 26 ›› Issue (11): 1714-1719.doi: 10.12307/2022.356

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Acupuncture combined with moxibustion regulates the expression of circadian clock protein in the synovium of rats with osteoarthritis

Tan Qian1, Li Bocun1, Li Jing1, Li Jia2, Xiang Hongchun1, Cai Guowei1   

  1. 1Department of Acupuncture and Moxibustion, Union Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China; 2College of Acupuncture and Orthopedics, Hubei University of Chinese Medicine, Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion, Wuhan 430061, Hubei Province, China
  • Received:2021-04-12 Revised:2021-04-15 Accepted:2021-06-01 Online:2022-04-18 Published:2021-12-11
  • Contact: Cai Guowei, Chief physician, Master’s supervisor, Department of Acupuncture and Moxibustion, Union Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • About author:Tan Qian, Master candidate, Department of Acupuncture and Moxibustion, Union Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81774410 (to CGW); Hubei Provincial Health Commission Project, No. ZY2021M018 (to CGW)

Abstract: BACKGROUND: Synovitis is one of the main pathological features of knee osteoarthritis. Previous studies have shown that acupuncture has a significant effect on knee osteoarthritis. Whether acupuncture can relieve the symptoms by regulating the related proteins in synovium is worthy of further study. 
OBJECTIVE: To investigate the effects of acupuncture combined with moxibustion on circadian clock proteins (Clock, Bmal1), TRPV4, and interleukin-1β in the synovium of rats with osteoarthritis. 
METHODS: Eighty 3-month-old male Sprague-Dawley rats were randomly divided into four groups: control group, model group, acupuncture group and acupuncture combined with moxibustion group (n=20 per group). Rat models of osteoarthritis were prepared by surgery in the latter three groups. Rats in the acupuncture group were treated with manual acupuncture at Zusanli (ST36), Dubi (ST35), and Neixiyan (EX-LE5) acupoints on the 7th day of modeling, at a depth of 5 mm, 15 minutes once a day, totally for 21 days. Rats in the acupuncture combined with moxibustion group were treated with acupuncture (same as that in the acupuncture group) combined with moxibustion. After corresponding treatments, the spontaneous activity ability of the rats was measured by an open field test. The pathological structure of the synovium was observed by hematoxylin-eosin staining. The expression of Clock, Bmal1 and transient receptor potential vanilloid 4 (TRPV4) in the synovium was detected by western blot. The expression of interleukin-1β in the synovium was detected by immunofluorescence labeling. 
RESULTS AND CONCLUSION: Compared with the control group, the expression of Clock and Bmal1 in the synovium was significantly decreased (P < 0.01), the levels of interleukin-1β and TRPV4 in the synovium were significantly increased (P < 0.05), and moreover severer loss of synovial epithelium was observed in the model group. Compared with the model group, acupuncture and acupuncture combined with moxibustion could significantly improve the spontaneous activity ability of the rats, increase the expression of Clock and Bmal1 (P < 0.05), decrease the levels of interleukin-1β and TRPV4 in the rat synovium  (P < 0.05), and relieve the loss of synovial epithelium. Compared with the acupuncture group, these changes were more obvious in the acupuncture combined with moxibustion group (P < 0.05). To conclude, relative to acupuncture alone, acupuncture combined with moxibustion can effectively regulate arthritic injury in knee osteoarthritis rats, reduce the expression of TRPV4 in synovial cells, promote the expression of Clock and Bmal1 in the synovium, thereby regulating the circadian clock in synovial cells, reducing the expression of inflammatory factors and slowing the degeneration of knee osteoarthritis.

Key words: acupuncture, Zusanli, knee osteoarthritis, circadian clock, Clock, Bmal1, TRPV4, interleukin-1β

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