Effects of acetazolamide on the proliferation , apoptosis , and inflammatory response of rheumatoid arthritis fibroblast⁃like synoviocytes by inhibiting autophagy
10. 19405/j. cnki. issn1000 - 1492. 2025. 12. 001
- VernacularTitle: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
- Author:
Mengqing Wang
1
;
Manyu Zhang
1
;
Shenglong Gu
1
;
Yan Huang
1
;
Rong Li
1
Author Information
1. School of Pharmacy, Anhui Medical University, Inflammation and Immune Mediated Diseases Laboratory of Anhui Province , Hefei 230032
- Publication Type:Journal Article
- Keywords:
acetazolamide;
rheumatoid arthritis;
fibroblast-like synoviocytes;
cell proliferation;
cell apoptosis;
autophagy
- From:
Acta Universitatis Medicinalis Anhui
2025;60(12):2187-2196
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effects and potential mechanisms of the aquaporin 1(AQP1) inhibitor acetazolamide(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 staining, 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 measure 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 decreased 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 cytoprotective autophagy , thereby alleviating the hyperproliferation , apoptosis resistance , and aberrant inflammatory response of RA⁃FLS , revealing the core role of autophagy inhibition in AZ ′s anti⁃RA effects.
- Full text:2026030223005029459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_王梦晴.pdf
