Abstract
Rheumatoid arthritis (RA) remains the most common inflammatory arthritis and a major cause of disability. This study investigated the mechanism of MLL3 in fibroblast-like synoviocyte (FLS) apoptosis and inflammatory factor secretion in RA. Expression of MLL3 in synovial tissue of RA patients and patients with bone trauma was detected. FLS was isolated and identified by flow cytometry. Expressions of TNF-α, IL-1β, IL-8, and IL-10 and apoptosis were measured by MTT, flow cytometry, and ELISA. Western blot and qRT-PCR were performed to detect MLL3 and CCL2 expressions, H3K4me3 level, and NF-κB pathway-related proteins in rat joints. MLL3 was highly expressed in the synovial tissue of RA patients, and silencing MLL3 in FLS-RA promoted apoptosis, inhibited pro-inflammatory factors TNF-α, IL-1β, and IL-8 secretion, and promoted anti-inflammatory factor IL-10 secretion. Inhibition of MLL3 suppressed intracellular H3K4me3 and CCL2 expressions. CCL2 activated the NF-κB pathway to promote pro-inflammatory factors TNF-α, IL-1β, and IL-8, inhibit anti-inflammatory factor IL-10, and inhibit apoptosis in FLS-RA. Inhibition of MLL3 expression in RA rats reduced joint redness, swelling, and intra-articular inflammation, but increasing H3K4me3 level reversed the ameliorative effects of sh-MLL3 on RA rats. Collectively, MLL3 activated the NF-κB pathway by increasing H3K4me3 modification in the CCL2 promoter region in FLS-RA, thereby inhibiting apoptosis and promoting pro-inflammatory factors of FLS-RA.
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The datasets and materials used during the current study are available from the corresponding author on reasonable request.
References
Grover, H.S., N. Gaba, A. Gupta, and C.M. Marya. 2011. Rheumatoid arthritis: a review and dental care considerations. Nepal Medical College Journal 13 (2): 74–76.
An, Q., W. Yan, Y. Zhao, and K. Yu. 2018. Enhanced neutrophil autophagy and increased concentrations of IL-6, IL-8, IL-10 and MCP-1 in rheumatoid arthritis. International Immunopharmacology 65: 119–128.
Scott, D.L., F. Wolfe, and T.W. Huizinga. 2010. Rheumatoid arthritis. Lancet 376 (9746): 1094–1108.
Smolen, J.S., D. Aletaha, and I.B. McInnes. 2016. Rheumatoid arthritis. Lancet 388 (10055): 2023–2038.
Ngian, G.S. 2010. Rheumatoid arthritis. Australian Family Physician 39 (9): 626–628.
Yang, B., Y. Ge, Y. Zhou, J. Wang, X. Xie, S. Li, M. Tang, L. Xu, and J. Tian. 2019. miR-124a inhibits the proliferation and inflammation in rheumatoid arthritis fibroblast-like synoviocytes via targeting PIK3/NF-kappaB pathway. Cell Biochemistry and Function 37 (4): 208–215.
Lee, S., J.S. Kong, S. You, H.M. Kwon, S.A. Yoo, C.S. Cho, and W.U. Kim. 2018. Transcription factor NFAT5 promotes migration and invasion of rheumatoid synoviocytes via coagulation factor III and CCL2. Journal of Immunology 201 (2): 359–370.
Koenders, M.I., and W.B. van den Berg. 2015. Novel therapeutic targets in rheumatoid arthritis. Trends in Pharmacological Sciences 36 (4): 189–195.
Sze, C.C., and A. Shilatifard. 2016. MLL3/MLL4/COMPASS Family on epigenetic regulation of enhancer function and cancer. Cold Spring Harbor Perspectives in Medicine 6 (11).
Lai, B., J.E. Lee, Y. Jang, L. Wang, W. Peng, and K. Ge. 2017. MLL3/MLL4 are required for CBP/p300 binding on enhancers and super-enhancer formation in brown adipogenesis. Nucleic Acids Research 45 (11): 6388–6403.
Araki, Y., Y. Aizaki, K. Sato, H. Oda, R. Kurokawa, and T. Mimura. 2018. Altered gene expression profiles of histone lysine methyltransferases and demethylases in rheumatoid arthritis synovial fibroblasts. Clinical and Experimental Rheumatology 36 (2): 314–316.
Chen, C.Y., L.J. Fuh, C.C. Huang, C.J. Hsu, C.M. Su, S.C. Liu, Y.M. Lin, and C.H. Tang. 2017. Enhancement of CCL2 expression and monocyte migration by CCN1 in osteoblasts through inhibiting miR-518a-5p: implication of rheumatoid arthritis therapy. Scientific Reports 7 (1): 421.
Lin, Y.C., Y.C. Lin, M.Y. Huang, P.L. Kuo, C.C. Wu, M.S. Lee, C.C. Hsieh, H.F. Kuo, C.H. Kuo, W.C. Tsai, and C.H. Hung. 2017. Tumor necrosis factor-alpha inhibitors suppress CCL2 chemokine in monocytes via epigenetic modification. Molecular Immunology 83: 82–91.
Han, P., D. Gao, W. Zhang, S. Liu, S. Yang, and X. Li. 2015. Puerarin suppresses high glucose-induced MCP-1 expression via modulating histone methylation in cultured endothelial cells. Life Sciences 130: 103–107.
Malemud, C.J. 2015. The PI3K/Akt/PTEN/mTOR pathway: a fruitful target for inducing cell death in rheumatoid arthritis? Future Medicinal Chemistry 7 (9): 1137–1147.
Li, J., R.S. Tang, Z. Shi, and J.Q. Li. 2020. Nuclear factor-kappaB in rheumatoid arthritis. International Journal of Rheumatic Diseases 23 (12): 1627–1635.
Duan, W., and H. Li. 2018. Combination of NF-kB targeted siRNA and methotrexate in a hybrid nanocarrier towards the effective treatment in rheumatoid arthritis. Journal of Nanobiotechnology 16 (1): 58.
Ogura, N., K. Satoh, M. Akutsu, M. Tobe, K. Kuyama, N. Kuboyama, H. Sakamaki, H. Kujiraoka, and T. Kondoh. 2010. MCP-1 production in temporomandibular joint inflammation. Journal of Dental Research 89 (10): 1117–1122.
Aletaha, D., T. Neogi, A.J. Silman, J. Funovits, D.T. Felson, C.O. Bingham, N.S. Birnbaum, G.R. Burmester, V.P. Bykerk, M.D. Cohen, B. Combe, K.H. Costenbader, M. Dougados, P. Emery, G. Ferraccioli, J.M. Hazes, K. Hobbs, T.W. Huizinga, A. Kavanaugh, J. Kay, T.K. Kvien, T. Laing, P. Mease, H.A. Menard, L.W. Moreland, R.L. Naden, T. Pincus, J.S. Smolen, E. Stanislawska-Biernat, D. Symmons, P.P. Tak, K.S. Upchurch, J. Vencovsky, F. Wolfe, and G. Hawker. 2010. 2010 rheumatoid arthritis classification criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Annals of the Rheumatic Diseases 69 (9): 1580–1588.
Klein, K., and S. Gay. 2015. Epigenetics in rheumatoid arthritis. Current Opinion in Rheumatology 27 (1): 76–82.
Xiong, W., H. Deng, C. Huang, C. Zen, C. Jian, K. Ye, Z. Zhong, X. Zhao, and L. Zhu. 2019. MLL3 enhances the transcription of PD-L1 and regulates anti-tumor immunity. Biochimica et Biophysica Acta - Molecular Basis of Disease 1865 (2): 454–463.
Zhao, J.X., H.X. Zhang, B. Li, Q.X. Fan, S.Z. Xue, M. Zhang, and C.Q. Ke. 2017. Role of MLL3 in regulating cardiac stem cells following cardiac cachexia. European Review for Medical and Pharmacological Sciences 21 (21): 4924–4929.
Quinones, M.P., C.A. Estrada, Y. Kalkonde, S.K. Ahuja, W.A. Kuziel, M. Mack, and S.S. Ahuja. 2005. The complex role of the chemokine receptor CCR2 in collagen-induced arthritis: implications for therapeutic targeting of CCR2 in rheumatoid arthritis. Journal of Molecular Medicine (Berlin, Germany) 83 (9): 672–681.
Liou, L.B., W.P. Tsai, C.J. Chang, W.J. Chao, and M.H. Chen. 2013. Blood monocyte chemotactic protein-1 (MCP-1) and adapted disease activity Score28-MCP-1: favorable indicators for rheumatoid arthritis activity. PLoS One 8 (1): e55346.
Kimball, A.S., A. Joshi, W.F. Carson 4th, A.E. Boniakowski, et al. 2017. The histone methyltransferase MLL1 directs macrophage-mediated inflammation in wound healing and is altered in a murine model of obesity and type 2 diabetes. Diabetes 66 (9): 2459–2471.
Sun, W., R. Qin, R. Qin, R. Wang, D. Ding, Z. Yu, Y. Liu, R. Hong, Z. Cheng, and Y. Wang. 2018. Sam68 promotes invasion, migration, and proliferation of fibroblast-like synoviocytes by enhancing the NF-kappaB/P65 pathway in rheumatoid arthritis. Inflammation 41 (5): 1661–1670.
Xu, J., H. Dong, Q. Qian, X. Zhang, Y. Wang, W. Jin, and Y. Qian. 2017. Astrocyte-derived CCL2 participates in surgery-induced cognitive dysfunction and neuroinflammation via evoking microglia activation. Behavioural Brain Research 332: 145–153.
Tang, S.C., P.Y. Liao, S.J. Hung, J.S. Ge, S.M. Chen, J.C. Lai, Y.P. Hsiao, and J.H. Yang. 2017. Topical application of glycolic acid suppresses the UVB induced IL-6, IL-8, MCP-1 and COX-2 inflammation by modulating NF-kappaB signaling pathway in keratinocytes and mice skin. Journal of Dermatological Science 86 (3): 238–248.
Wu, M., J. Gu, S. Mei, D. Xu, Y. Jing, Q. Yao, M. Chen, M. Yang, S. Chen, B. Yang, N. Qi, H. Hu, R.P. Wüthrich, and C. Mei. 2016. Resveratrol delays polycystic kidney disease progression through attenuation of nuclear factor kappaB-induced inflammation. Nephrology, Dialysis, Transplantation 31 (11): 1826–1834.
Wang, L., N. Wang, Q. Zhao, B. Zhang, and Y. Ding. 2019. Pectolinarin inhibits proliferation, induces apoptosis, and suppresses inflammation in rheumatoid arthritis fibroblast-like synoviocytes by inactivating the phosphatidylinositol 3 kinase/protein kinase B pathway. Journal of Cellular Biochemistry 120 (9): 15202–15210.
Meng, Q., X. Du, H. Wang, H. Gu, J. Zhan, and Z. Zhou. 2017. Astragalus polysaccharides inhibits cell growth and pro-inflammatory response in IL-1beta-stimulated fibroblast-like synoviocytes by enhancement of autophagy via PI3K/AKT/mTOR inhibition. Apoptosis 22 (9): 1138–1146.
Acknowledgements
We thank all the members in our team for the excellent work.
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This research was supported by funds from the 2019 Xinxiang City Science and Technology Research Plan Project(GG2019026); The Henan Medical Science and Technology Public Relations Plan jointly established a project(LHGJ20200959, LHGJ20200941).
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Conceptualization: WF and SL. Validation, research, resources, data reviewing, and writing: ZX, AZ, and ZB. Review and editing: XG, YQ, and JW. All authors read and approved the final manuscript.
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This study obtained approval from the ethics committee of Xinxiang Central Hospital, and all procedures were conducted according to the declaration of Helsinki. The animal experiments were performed with the principle of minimized animal number and the least pains.
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Fan, W., Xu, Z., Liang, S. et al. MLL3 Inhibits Apoptosis of Rheumatoid Arthritis Fibroblast-Like Synoviocytes and Promotes Secretion of Inflammatory Factors by Activating CCL2 and the NF-κB Pathway. Inflammation 44, 1803–1814 (2021). https://doi.org/10.1007/s10753-021-01459-2
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DOI: https://doi.org/10.1007/s10753-021-01459-2