ALKBH5-YTHDF2 m6A 修饰轴通过抑制 NLRP3 抑制类风湿关节炎进展。

ALKBH5-YTHDF2 m6A modification axis inhibits rheumatoid arthritis progression by suppressing NLRP3.

机构信息

Department of Rheumatology and Immunology, Shenzhen Futian Hospital for Rheumatic Diseases, Shenzhen, 518000, PR China.

Department of Rheumatology, Shanghai Guanghua Hospital of Integrated Traditional and Western Medicine, Shanghai, 200052, China.

出版信息

Biochem Biophys Res Commun. 2023 Aug 6;668:70-76. doi: 10.1016/j.bbrc.2023.05.087. Epub 2023 May 23.

DOI:10.1016/j.bbrc.2023.05.087

PMID:37244037

Abstract

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease. Recently, NLRP3 has been demonstrated to be closely related to RA. The objective of our research was to analyze the specific mechanism of NLRP3 in RA. The m6A levels of NLRP3 was detected with methylated RNA immunoprecipitation (MeRIP) kit. The mRNA and protein levels of related genes were tested with RT-qPCR and Western blot. The inflammatory factors levels were detected with ELISA kits. The cell proliferative ability was measured with CCK-8 and EdU staining assays. NLRP3 levels was prominently in synovial tissues and fibroblast-like synoviocytes (FLS) from RA patients. NLRP3 silencing suppressed FLS proliferation and inflammatory factor levels. Additionally, ALKBH5 was found to bind with NLRP3, and ALKBH5 silencing suppressed FLS proliferation and inflammatory factor levels while NLRP3 overexpressing neutralized the role of ALKBH5 in FLS. Furthermore, m6A modified induced by ALKBH5 suppressed NLRP3 mRNA level through YTHDC2 in RA, and NLRP3 is a hinge factor in RA progression.

摘要

类风湿关节炎（RA）是一种慢性炎症性自身免疫性疾病。最近，NLRP3 被证明与 RA 密切相关。我们的研究目的是分析 NLRP3 在 RA 中的具体作用机制。采用甲基化 RNA 免疫沉淀（MeRIP）试剂盒检测 NLRP3 的 m6A 水平。采用 RT-qPCR 和 Western blot 检测相关基因的 mRNA 和蛋白水平。采用 ELISA 试剂盒检测炎性因子水平。通过 CCK-8 和 EdU 染色检测细胞增殖能力。NLRP3 在 RA 患者的滑膜组织和成纤维样滑膜细胞（FLS）中水平明显升高。NLRP3 沉默抑制了 FLS 的增殖和炎性因子水平。此外，发现 ALKBH5 与 NLRP3 结合，ALKBH5 沉默抑制了 FLS 的增殖和炎性因子水平，而 NLRP3 的过表达中和了 ALKBH5 在 FLS 中的作用。此外，ALKBH5 诱导的 m6A 修饰通过 YTHDC2 抑制 RA 中 NLRP3 mRNA 水平，而 NLRP3 是 RA 进展的枢纽因素。