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Effect of Electroacupuncture in “Zusanli” and “Kunlun” Acupoints on TLR4 Signaling Pathway of Adjuvant Arthritis Rats

Dong, Zhe-Qing MD1,*; Zhu, Ji MD1; Lu, De-zhao PhD2; Chen, Qin PhD1; Xu, Ying-ling MD2

American Journal of Therapeutics: May/June 2018 - Volume 25 - Issue 3 - p e314–e319
doi: 10.1097/MJT.0000000000000477
Original Article

Background: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Previous study suggested that toll-like receptor (TLR) signaling pathway contributes to the development and progression of RA. In recent years, acupuncture has become one of the most vital treatments of arthralgia. But little is known about the mechanisms of improving RA by acupuncture.

Study Question: The study studied the effect of electroacupuncture in “Zusanli” and “Kunlun” acupoints on the expression of TLR4, myeloid differentiation factor 88 (MYD88), and NF-κB in adjuvant arthritis rats to clarify the molecular mechanism of acupuncture of RA.

Study Design: A rat model of adjuvant arthritis was established with injection of 0.1 mL Freund complete adjuvant in the right hindlimb footpad. We next punctured the Zusanli and Kunlun acupoints with 0.25 × 40-mm acupuncture needles to 5-mm depth. Then, we performed electroacupuncture treatment for 28 days with frequency of 2 Hz and intensity of 2 mA, once a day and 30 minutes each time.

Measures and Outcomes: Arthritis index and paw swelling were measured every week. FQ-PCR and western blot were used to detect the expression of TLR4, MYD88, and NF-κB.

Results: Paw swelling of rats injected with Freund complete adjuvant was more serious than that of the normal rats, which illustrated the successful establishment of adjuvant arthritis rat model. After treatment for 14 days, the paw swelling and joint symptoms score decreased, paw tissue inflammation eased in the rats of treatment group compared with the model group during the same period. After treatment for 28 days, the expression of TLR4, MYD88, and NF-κB in the ankle bone tissues decreased at both mRNA and protein levels.

Conclusions: Stimulation with electric needle in Zusanli and Kunlun acupoints can reduce the expression of TLR4, MYD88, and NF-κB, which play an important role in treatment of adjuvant arthritis.

1Clinical Laboratory, The Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, China; and

2College of Life Science, Zhejiang Chinese Medical University, Hangzhou, China.

Address for correspondence: Clinical Laboratory, The Third Affiliated Hospital of Zhejiang Chinese Medical University, No. 219, Mogan Road, Hangzhou 310005, Zhejiang, China. E-mail:

Supported by Zhejiang Provincial Natural Science Foundation of China (Y211159), Research Foundation of Traditional Chinese Medicine of Zhejiang Province, China (2013ZA066), Research Foundation of Education Bureau of Zhejiang Province, China (Y201017345).

The authors have no conflicts of interest to declare.

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