海南医科大学学报2025，Vol.31Issue(2)：94-101,8.DOI:10.13210/j.cnki.jhmu.20241106.002
基于METTL14-m6A-FOXO3A信号通路介导的细胞自噬探讨补肾强督方抑制炎症治疗强直性脊柱炎的机制研究
Bushen Qiangdu formula in suppressing inflammation in ankylosing spondylitis via METTL14-m6A-FOXO3A signaling pathway mediated autophagy
摘要
Abstract
Objective:To investigate the effect of Bushen Qiangdu formula on METTL14-m6A-FOXO3A signaling pathway-mediated autophagy in T lymphocytes,and to elucidate its molecular mechanism in suppressing inflammation for the treatment of ankylosing spondylitis(AS).Methods:Jurkat cells were stimulated with anti-CD3 and anti-CD28 antibodies for 24 h to establish the cell model,followed by intervention with Bushen Qiangdu medicated serum and METTL14 recombinant protein.ELISA was used to measure the levels of inflammatory factors(IL-23 and IL-17A)and oxidative stress markers.ROS levels were assessed by flow cytometry.Western blot methodology was utilized to ascertain the levels of expression of autophagy-related proteins,namely METTL14 and FOXO3A.Real-time quantitative polymerase chain reaction was used to determine the mRNA and protein expres-sion levels of METTL14 and FOXO3A,and dot blot hybridization was performed to assess the m6A methylation level of FOXO3A.Results:Compared to the model group,Bushen Qiangdu medicated serum significantly reduced the levels of IL-23,IL-17A,and MDA,as well as ROS levels of Jurkat cells,all of which were statistically significant(P<0.01),while the activity of SOD and GSH-Px were increased.The results demonstrated statistically significant discrepancies between the experimental and control groups(P<0.01).Additionally,Bushen Qiangdu medicated serum notably upregulated the expression of Beclin-1,FOXO3A,and METTL14 proteins,and increased the LC3II/LC3I ratio while downregulating p62 protein expression(P<0.01).Moreover,Bushen Qiangdu medicated serum significantly enhanced the m6A methylation level of FOXO3A in Jurkat cells compared to the model group(P<0.01).Conclusion:Bushen Qiangdu formula promotes T cell autophagy through METTL14-mediated m6A methylation of FOXO3A,achieving therapeutic effects in AS.关键词
补肾强督方/T淋巴细胞/METTL14/FOXO3A/m6A甲基化修饰Key words
Bushen Qiangdu formula/T lymphocytes/METTL14/FOXO3A/m6A methylation modification分类
医药卫生引用本文复制引用
吴琳,李松倍,苏晓庆,韩天然,李泽光..基于METTL14-m6A-FOXO3A信号通路介导的细胞自噬探讨补肾强督方抑制炎症治疗强直性脊柱炎的机制研究[J].海南医科大学学报,2025,31(2):94-101,8.基金项目
This study was supported by the Bethune Medical Science Research Fund(TY144EN) 白求恩医学科学研究基金(TY144EN) （TY144EN）
