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Hypoxia and TNF-α Synergistically Induce Expression of IL-6 and IL-8 in Human Fibroblast-like Synoviocytes via Enhancing TAK1/NF-κB/HIF-1α Signaling

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Abstract—

Hypoxia and increased levels of inflammatory cytokines in the joints are characteristics of rheumatoid arthritis (RA). However, the effects of hypoxia and tumor necrosis factor-α (TNF-α) on interleukin (IL)-6 and IL-8 production on fibroblast-like synoviocytes (FLSs) remain to be clarified. This study aimed to explore how hypoxia and TNF-α affect the expression of IL-6 and IL-8 in human FLSs isolated from RA patients. Hypoxia or TNF-α treatment alone significantly increased the expression and promoter activity of IL-6, IL-8, and hypoxia-inducible factor-1α (HIF-1α). Treatment of hypoxic FLSs with TNF-α further significantly elevated the expression of these cytokines and enhanced promoter activity of HIF-1α, which was abrogated by treatment with the HIF-1α inhibitor YC-1. Similarly, TNF-α alone elevated the phosphorylation and promoter activity of nuclear factor-κBp65 (NF-κBp65) in the FLSs. These effects were further enhanced by the combined treatment of hypoxia and TNFα but were attenuated by the NF-κB inhibitor BAY11-7082. NF-κB-p65 inhibition decreased the effect of TNF-α on HIF-1α upregulation in the FLSs in response to hypoxia. The combination of hypoxia and TNF-α also significantly upregulated transforming growth factor-β-activated kinase 1 (TAK1) expression, and silencing TAK1 dramatically decreased NF-κB-p65, HIF-1α, IL-6, and IL-8 expression under the same conditions. Our results indicate that hypoxia and TNF-α synergistically increase IL-6 and IL-8 expression in human FLSs via enhancing TAK1/NF-κB/HIF-1α signaling.

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Funding

This work was supported by grants from the Joint Funds of the Zhejiang Provincial Natural Science Foundation of China (Grant number LTY21H100001) and the College Students’ Innovation and Entrepreneurship Training Program Project of China (Grant number 202110350052,202210350058).

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Authors and Affiliations

  1. Department of Rheumatology and Immunology, Taizhou Central Hospital of Taizhou University, Taizhou, 318000, Zhejiang, China

    Guofen Wang, Junsong Wang, Xian Li & Xinjing Luo

  2. Department of Basic Medical Science, School of Medicine of Taizhou University, Taizhou, 318000, Zhejiang, China

    Qiyang Wu, Ruifeng Yao & Xinjing Luo

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  1. Guofen Wang
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  2. Junsong Wang
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  3. Xian Li
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Contributions

W.G. designed the experiments and edited the final manuscript. W. J. and L. X. collected synovial tissue samples and analyzed the data. W. Q. and Y. R. performed the experiments and data analysis. L. X. conceived and designed the study and drafted the manuscript. All authors reviewed and approved the final manuscript.

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Correspondence to Xinjing Luo.

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The experiments were performed according to the protocols approved by the Ethics Committee of the Affiliated Hospital of Taizhou University (2020-sc-032).

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Wang, G., Wang, J., Li, X. et al. Hypoxia and TNF-α Synergistically Induce Expression of IL-6 and IL-8 in Human Fibroblast-like Synoviocytes via Enhancing TAK1/NF-κB/HIF-1α Signaling. Inflammation 46, 912–924 (2023). https://doi.org/10.1007/s10753-022-01779-x

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