Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Inhibition of miR-665 alleviates lipopolysaccharide-induced inflammation via up-regulation of SOCS7 in chondrogenic ATDC5 cells

Qiaoyi Ning^1,2, Yiting He³ , Wukai Ma², Fang Tang², Ying Huang², Xueming Yao²

¹The 2nd Clinical School, Guangzhou University of Chinese Medicine, Guangzhou City, Guangdong Province 510120; ²Department of Rheumatology and Immunology, The 2nd hospital affiliated to Guizhou University of Chinese Traditional Medicine, Guiyang City, Guizhou Province 550003; ³Department of New Drug Development, the 2nd Clinical School, Guangzhou University of Chinese Medicine, Guangzhou City, Guangdong Province 510120, China.

For correspondence:- Yiting He Email: YitingHedjk@163.com Tel:+862081887233

Accepted: 24 September 2020 Published: 30 October 2020

Citation: Ning Q, He Y, Ma W, Tang F, Huang Y, Yao X. Inhibition of miR-665 alleviates lipopolysaccharide-induced inflammation via up-regulation of SOCS7 in chondrogenic ATDC5 cells. Trop J Pharm Res 2020; 19(10):2067-2072 doi: 10.4314/tjpr.v19i10.X7

© 2020 The authors.
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Abstract

Purpose: To examine the e?ect and mechanism of action of miR-665 in osteoarthritis.

Methods: An in vitro inflammatory injury model of osteoarthritis was established using chondrogenic ATDC5 cells with lipopolysaccharide (LPS) treatment. The expression levels of inflammatory cytokines were determined by enzyme-linked immunosorbent assays (ELISAs) and by quantitative real-time polymerase chain reaction (qRT-PCR). A binding target for miR-665 was predicted using TargetScan and then evaluated using a dual-luciferase reporter assay.

Results: Treatment with LPS significantly up-regulated the inflammatory cytokine expressions of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-alpha (TNF-α), in ATDC5 cells (p < 0.01), and the expression of miRNA-665 was significantly increased in LPS-treated ATDC5 cells (p < 0.01). Knockdown of miR-665 down-regulated the expression levels of these inflammatory cytokines. Suppressor of cytokine signaling 7 (SOCS7) was identified as a target of miR-665. Data from qRT-PCR and western-blot analyses indicated that SOCS7 expression was promoted by miR-665 inhibition and inhibited by miR-665 over-expression. LPS treatment significantly decreased the expression of SOCS7 protein in ATDC5 cells (p < 0.01), and over-expression of SOCS7 attenuated the LPS-stimulated inflammatory injury. In addition, over-expression of miR-655 enhanced the inflammatory injury and reversed the protective e?ect of SOCS7 against LPS-stimulated inflammation.

Conclusion: Inhibition of miR-665 alleviated LPS-stimulated inflammatory injury in ATDC5 cells via the up-regulation of SOCS7, suggesting a potential therapeutic target for osteoarthritis.

Keywords: MiR-665, Lipopolysaccharide, Inflammation, SOCS7, Chondrogenic, ATDC5