Effect of Wutou Decoction on TLR4/NF-κB signaling pathway in synovial tissue of rat models with knee osteoarthritis

doi:10.3969/j.issn.2095-4344.1389

Abstract

Abstract:

BACKGROUND: Preliminary study has shown that Wutou Decoction can effectively inhibit inflammatory reaction in knee osteoarthritis. But its underling mechanism remains unclear.
OBJECTIVE: To observe the effect of Wutou Decoction on the related regulatory factors in TLR4/NF-κB signaling pathway in synovium of rats with knee osteoarthritis, so as to explore its anti-inflammatory mechanism on synovitis of knee osteoarthritis.
METHODS: Thirty-six male Sprague-Dawley rats, SPF grade, were provided by Shanghai Slack Laboratory Animals Co., Ltd., and the study was approved by the Experimental Animal Ethics Committee of Fujian University of Traditional Chinese Medicine. After 1 week of acclimation, the rats were randomly divided into blank, model and treatment groups. The rats in the model and treatment groups were used for establishing osteoarthritis model in both knees by modified Hulth method. One week later, the blank and model groups received 10 mL/(kg•d) saline via gavage. The experiment group received 4.2 g/(kg•d) Wutou Decoction. After 8 weeks of treatment, the rat synovial tissue was obtained. The contents of inducible nitric oxide synthase, tumor necrosis factor-α and interleukin-6 were measured by ELISA. The mRNA expression levels of TLR4, MyD88, TRAF6 and NF-κB p65 were measured by real-time PCR. The protein levels of TLR4, MyD88, TRAF6 and NF-κB p65 were measured by western blot assay.
RESULTS AND CONCLUSION: (1) ELISA results showed that Wutou Decoction could effectively inhibit the expression of inducible nitric oxide synthase, tumor necrosis factor-α and interleukin-6 compared with the model group (P < 0.01). (2) Real-time PCR results suggested that Wutou Decoction could inhibit the mRNA expression of TLR4, MyD88, TRAF6 and NF-κB p65 compared with the model group (P < 0.01). (3) The results of western blot assay were in agreement with the results of real-time PCR. That was, Wutou Decoction could inhibit the protein expression of TLR4, MyD88, TRAF6 and NF-κB p65 (P < 0.01). (4) These results suggest that Wutou Decoction can inhibit the inflammatory reaction in synovitis of knee osteoarthritis by regulating TLR4/NF-κB signaling pathway, which may, in part, explain its clinical efficacy in the treatment of synovitis of knee osteoarthritis.

Key words: Wutou Decoction, knee osteoarthritis, synovitis, TLR4/NF-κB signaling pathway, TLR4, MyD88, inducible nitric oxide synthase, tumor necrosis factor-α, interleukin-6

CLC Number:

Chen Jun1, Lin Jie2, Zhao Zhongsheng2, Huang Yanfeng2, Wu Guangwen3. Effect of Wutou Decoction on TLR4/NF-κB signaling pathway in synovial tissue of rat models with knee osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(27): 4381-4386.

Figures/Tables 1

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