MiR-146a Upregulates the TLR4/NF-κb Signaling Pathway to Promote Cytokine Expression and Synovial Fibroblast Proliferation in Rheumatoid Arthritis

Yuan-hao Wu^1,2, Wei Liu¹, Lei Zhang³, Bin Xue³, Yi Wang³, Xiao-ya Liu³, Yang Ji⁴, Ran Duan³, Yue Cai³ and Bo Zhang³, ¹Department of Rheumatology and Immunology, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China, ²Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing, China, ³First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China, ⁴The 272nd Hospital of Chinese People's Liberation Army, Tianjin, China

Meeting: 2017 ACR/ARHP Annual Meeting

Date of first publication: September 18, 2017

Keywords: Cell Signaling, MicroRNA, NFB, rheumatoid arthritis, synovium and toll-like receptors

Session Information

Date: Tuesday, November 7, 2017

Title: Rheumatoid Arthritis – Human Etiology and Pathogenesis Poster III

Session Type: ACR Poster Session C

Session Time: 9:00AM-11:00AM

Background/Purpose: To investigate the role of miR-146a in the activation of toll-like receptor-4 (TLR4)/ nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway, and the effect of miR-146a in cytokines expression and synoviocytes proliferation during rheumatoid arthritis (RA).

Methods: Synovial tissues were collected from RA patients who received arthroscopic synovectomy in the Department of Rheumatism at our hospital. Cells were categorized into a blank control group, a negative control group, a miR-146a mimics group and a miR-146a inhibitors group. The cholecystokinin (CCK)-8 assay was used to measure the inhibition on cell proliferation rate. TLR4 mRNA level, as well as the mRNA expressions of NF-κB, interleukin (IL)-1β, IL-6, IL-8, IL-17, cyclooxygenase (COX)-2, matrix metalloproteinase (MMP)-3, Seprase, and inducible nitric oxide synthase (iNOS) were detected by the reverse transcription-polymerase chain reaction (RT-PCR). Western blot was used to measure TLR4 and NF-κB proteins, and the expressions of IL-1β, IL-6, IL-8, IL-17, Seprase, MMP-3 and prostaglandin E2 (PGE2) were measured by enzyme linked immunosorbent assay (ELISA). The content of nitric oxide (NO) was measured by the Griess method.

Results: The CD3 and CD14 expression rate gradually decreased, and the CD90 expression rate gradually increased. After the third generation of cell culture, the CD90 expression rate (96.48%) was significantly higher than those of CD3 (1.17%) and CD14 (1.27%). After culturing for 48h and 72h, cell proliferation in the miR-146a mimics group was significantly increased (both P < 0.05) as compared with the blank group, while that in the miR-146a inhibitors group significantly reduced (both P < 0.05). With an extended time of cell culture, the change in cell proliferation was more significant, and differences were observed at each time point (all P < 0.05). Compared with the blank group, the expressions of TLR4, NF-κB, IL-1, IL-6, IL-8, IL-17, COX-2, MMP-3, Seprase, iNOS and NO content in the miR-146a mimics group increased significantly (all P < 0.05), but in the miR-146a inhibitors group, these values significantly reduced (all P < 0.05).

Conclusion: The inhibition of miR-146a expression could block the TLR4/NF-κB signaling pathway, and inhibited synovial fibroblast proliferation and cytokine expressions in RA.

Disclosure: Y. H. Wu, None; W. Liu, None; L. Zhang, None; B. Xue, None; Y. Wang, None; X. Y. Liu, None; Y. Ji, None; R. Duan, None; Y. Cai, None; B. Zhang, None.

To cite this abstract in AMA style:

Wu YH, Liu W, Zhang L, Xue B, Wang Y, Liu XY, Ji Y, Duan R, Cai Y, Zhang B. MiR-146a Upregulates the TLR4/NF-κb Signaling Pathway to Promote Cytokine Expression and Synovial Fibroblast Proliferation in Rheumatoid Arthritis [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/mir-146a-upregulates-the-tlr4nf-%ce%bab-signaling-pathway-to-promote-cytokine-expression-and-synovial-fibroblast-proliferation-in-rheumatoid-arthritis/. Accessed .

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