m⁶A-mediated lncRNA MAPKAPK5-AS1 induces apoptosis and suppresses inflammation via regulating miR-146a-3p/SIRT1/NF-κB axis in rheumatoid arthritis

ABSTRACT

To investigate the role of m⁶A-mediated lncRNA MAPKAPK5-AS1 (MK5-AS1) in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLSs) and its underlying molecular mechanism. RT-qPCR, western blot, flow cytometry (FCM), and enzyme-linked immunosorbent assay (ELISA) were utilized for evaluating inflammation and apoptosis. Next, RIP, RNA pull-down, dual-luciferase reporter gene assay, and a series of rescue experiments were performed to explore the regulatory mechanisms of MK5-AS1 and its sponge-like action in RA-FLSs. The regulatory relationships between MK5-AS1 and WTAP were explored using the MeRIP-qPCR assay and RT-qPCR. Finally, the critical RNAs in the ceRNA axis were verified in the clinical cohort. MK5-AS1 was poorly expressed and miR-146a-3p was overexpressed in co-cultured RA-FLSs. MK5-AS1 overexpression could inhibit inflammatory responses and promote cell apoptosis in the co-cultured RA-FLSs. MK5-AS1 bound to miR-146a-3p to target SIRT1, thereby affecting inflammatory responses and cell apoptosis in the co-cultured RA-FLSs. SIRT1 knockdown or miR-146a-3p overexpression reversed the impacts of MK5-AS1 overexpression on co-cultured RA-FLSs inflammation and apoptosis. Moreover, WTAP was downregulated, and induced the inhibition of MK5-AS1 by promoting its RNA transcript stability. Clinically, MK5-AS1 was downregulated in RA-PBMCS and correlated with the clinical characteristics of RA. Our study elucidated that m⁶A-mediated MK5-AS1 sequestered miR-146a-3p to suppress SIRT1 expression in co-cultured RA-FLSs, thus providing a new insight into the treatment of rheumatoid arthritis.

KEYWORDS:

• Rheumatoid arthritis
• MAPKAPK5-AS1
• ceRNA
• m⁶A
• apoptosis
• inflammatory responses

Author contributions

LJ, WL, JH, and WJT conceived and designed the experiment. WJT, WX, and WJ performed most of the experiments. SY and WL contributed to the patient samples. WJT and SY analyzed the statistical analyses. XL and FYY guidelined to how to proceed with clinical data mining. WJT wrote the manuscript. All authors revised the manuscript. All authors contributed to the article and approved the submitted version.

Disclosure statement

No potential conflict of interest was reported by the authors.

Data availability statement

All relevant data and materials are stored at the Anhui Province Key Laboratory of Modern Chinese Medicine Department of Internal Medicine Application Foundation Research and Development and can be obtained from the first author and corresponding author.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/15384101.2024.2302281

Additional information

Funding

This work was supported by grants from the Ministry of Science and Technology National Key Research and Development Program Chinese Medicine Modernization Research Key Project (2018YFC1705204); National Nature Fund Program (82274490, 81973655, 82074373); The University Synergy Innovation Program of Anhui Province (GXXT-2020-025); Open Foundation of Key Laboratory of Xin’an Medical Ministry of Anhui University of Traditional Chinese Medicine (No.2020×ayx10); Open Foundation of Anhui Province Key Laboratory of Modern Chinese Medicine Department of Internal Medicine Application Foundation Research and Development (2021AKLMCM004); Anhui Province Major and Intractable Diseases Collaborative Research Project of Traditional Chinese and Western Medicine (Anhui Traditional Chinese Medicine Development [2021] No. 70).