安徽医科大学学报2025，Vol.60Issue(12)：2187-2196,10.DOI:10.19405/j.cnki.issn1000-1492.2025.12.001
乙酰唑胺抑制自噬对类风湿关节炎成纤维样滑膜细胞增殖、凋亡和炎症反应的影响
Effects of acetazolamide on the proliferation,apoptosis,and inflammatory response of rheumatoid arthritis fibroblast-like synoviocytes by inhibiting autophagy
摘要
Abstract
Objective To study the effects and potential mechanisms of the aquaporin 1(AQP1)inhibitor acetazo-lamide(AZ)on the proliferation,apoptosis,and inflammatory response of rheumatoid arthritis(RA)fibroblast-like synoviocytes(FLS).Methods TNF-α-induced RA-FLS was served as in vitro RA model.MTT assay,IF staining,and EdU incorporation assay were applied to study AZ's effects on RA-FLS proliferation.Hoechst stai-ning,flow cytometry analysis of Annexin V-FITC/PI-stained cells,and mitochondrial membrane potential detection experiments were used to detect cell apoptosis.ELISA and quantitative real-time PCR methods were used to meas-ure pro-inflammatory cytokine production.Cell autophagy was evaluated using IF staining and mCherry-GFP-LC3B puncta assay.Western blot was performed to detect the levels of autophagy,apoptosis,and proliferation-related proteins.Moreover,the role of autophagy inhibition in AZ's effects on RA-FLS was examined by co-treating with the autophagy activator rapamycin(RAPA)or the autophagy inhibitor 3-methyladenine(3-MA).Results AZ dose-dependently inhibited cell proliferation,promoted apoptosis,and reduced the production of pro-inflammatory cytokines in RA-FLS.Furthermore,AZ suppressed cytoprotective autophagy in these cells,as evidenced by de-creased LC3B levels(P＜0.05),increased p62 expression(P＜0.05),and reduced autophagic flux(P＜0.01).Particularly,AZ's beneficial effects were reversed by RAPA-induced autophagy activation and enhanced by 3-MA-induced autophagy inhibition.Conclusion This study provides the first evidence that AZ hinders cytopro-tective autophagy,thereby alleviating the hyperproliferation,apoptosis resistance,and aberrant inflammatory re-sponse of RA-FLS,revealing the core role of autophagy inhibition in AZ's anti-RA effects.关键词
乙酰唑胺/类风湿关节炎/成纤维样滑膜细胞/细胞增殖/细胞凋亡/自噬Key words
acetazolamide/rheumatoid arthritis/fibroblast-like synoviocytes/cell proliferation/cell apoptosis/autophagy分类
医药卫生引用本文复制引用
王梦晴,张曼玉,顾胜龙,黄艳,李荣..乙酰唑胺抑制自噬对类风湿关节炎成纤维样滑膜细胞增殖、凋亡和炎症反应的影响[J].安徽医科大学学报,2025,60(12):2187-2196,10.基金项目
国家自然科学基金项目(编号:81972040) （编号:81972040）
安徽省高校协同创新项目(编号:GXXT-2022-063) National Natural Science Foundation of China(No.81972040) （编号:GXXT-2022-063）
Collaborative Innovation Pro-ject of Anhui Province Universities(No.GXXT-2022-063) （No.GXXT-2022-063）
