Xiong Xiaochuan, Xiong Hao, Peng Jun, Liu Yingjie, Zong Yang
Department of Orthopaedics, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai, China.
Cartilage. 2025 Mar;16(1):89-99. doi: 10.1177/19476035231200336. Epub 2023 Sep 19.
Osteoarthritis (OA) is a common degenerative joint disease. The occurrence of OA slowly destroys the soft tissue structure of the patient's joint. Severe cases could lead to disability. Current studies had shown that inhibition of chondrocytes pyroptosis could slow down the progression of OA. Our work aimed to explore the specific mechanisms and ways of regulating this process.
In this work, the level of N-methyladenosine (mA) in clinical tissues was detected by ribonucleic acid (RNA) mA dot blot. qRT-PCR (quantitative real-time polymerase chain reaction) was used to detect the messenger RNA (mRNA) expression level of mA modified enzyme in clinical tissues. MTT (3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromid) and flow cytometry were used to detect the effect of sh-METTL3 (methyltransferase like 3) and NIMA-related kinase 7 (NEK7) transfection on chondrocytes pyroptosis in OA. Western blot was used to detect the protein expression levels of pyroptosis-related proteins. ELISA (enzyme-linked immunosorbent assay) was used to measure the protein concentration of inflammatory cytokines. The SRAMP online database was used to predict the mA site of NEK7. HE staining was used to assess the progression of OA in mice.
The level of mA in clinical samples of OA patients was higher, and METTL3 was significantly higher expressed in clinical samples of OA patients. We provided evidence that low expression of METTL3 inhibited chondrocytes pyroptosis. In addition, Rescue experiments and experiments had shown that METTL3 in combination with NEK7 inhibited the progression of OA by promoting chondrocytes pyroptosis.
METTL3 regulates mA modification of NEK7 and inhibits OA progression.
骨关节炎（OA）是一种常见的退行性关节疾病。OA的发生会缓慢破坏患者关节的软组织结构。严重病例可能导致残疾。目前的研究表明，抑制软骨细胞焦亡可减缓OA的进展。我们的工作旨在探索调节这一过程的具体机制和方法。
在本研究中，通过核糖核酸（RNA）甲基化斑点印迹法检测临床组织中N-甲基腺苷（mA）的水平。采用qRT-PCR（定量实时聚合酶链反应）检测临床组织中mA修饰酶的信使核糖核酸（mRNA）表达水平。使用MTT（3-(4,5)-二甲基噻唑-2,5-二苯基四氮唑溴盐）和流式细胞术检测sh-METTL3（甲基转移酶样3）和NIMA相关激酶7（NEK7）转染对OA中软骨细胞焦亡的影响。采用蛋白质免疫印迹法检测焦亡相关蛋白的表达水平。酶联免疫吸附测定（ELISA）用于检测炎性细胞因子的蛋白浓度。利用SRAMP在线数据库预测NEK7的mA位点。采用苏木精-伊红（HE）染色评估小鼠OA的进展情况。
OA患者临床样本中mA水平较高，且METTL3在OA患者临床样本中的表达显著升高。我们提供的证据表明，METTL3低表达可抑制软骨细胞焦亡。此外，挽救实验表明，METTL3与NEK7联合通过促进软骨细胞焦亡抑制OA的进展。
METTL3调节NEK7的mA修饰并抑制OA进展。
