Ca 2+ signalling plays a role in celastrol-mediated suppression of synovial fibroblasts of rheumatoid arthritis patients and experimental arthritis in rats

• Published in: British journal of pharmacology 2019-08, Vol.176 (16), p.2922-2944
• Main Authors: Wong, Vincent Kam Wai, Qiu, Congling, Xu, Su-Wei, Law, Betty Yuen Kwan, Zeng, Wu, Wang, Hui, Michelangeli, Francesco, Dias, Ivo Ricardo De Seabra Rodrigues, Qu, Yuan Qing, Chan, Tsz Wai, Han, Yu, Zhang, Ni, Mok, Simon Wing Fai, Chen, Xi, Yu, Lu, Pan, Hudan, Hamdoun, Sami, Efferth, Thomas, Yu, Wen Jing, Zhang, Wei, Li, Zheng, Xie, Yuesheng, Luo, Riqiang, Jiang, Quan, Liu, Liang
• Format: Article
• Language: English
• Subjects: Animals; Arthritis, Experimental - drug therapy; Arthritis, Experimental - metabolism; Arthritis, Rheumatoid - drug therapy; Arthritis, Rheumatoid - metabolism; Autophagy - drug effects; Calcium Signaling - drug effects; Cells, Cultured; Fibroblasts - drug effects; Fibroblasts - metabolism; Gene Expression Regulation - drug effects; Humans; Male; Mice, Knockout; Pentacyclic Triterpenes; Rats, Sprague-Dawley; Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism; Synovial Membrane - cytology; Triterpenes - pharmacology; Triterpenes - therapeutic use
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/bph.14718

Celastrol exhibits anti-arthritic effects in rheumatoid arthritis (RA), but the role of celastrol-mediated Ca mobilization in treatment of RA remains undefined. Here, we describe a regulatory role for celastrol-induced Ca signalling in synovial fibroblasts of RA patients and adjuvant-induced arthritis (AIA) in rats. We used computational docking, Ca dynamics and functional assays to study the sarcoplasmic/endoplasmic reticulum Ca ATPase pump (SERCA). In rheumatoid arthritis synovial fibroblasts (RASFs)/rheumatoid arthritis fibroblast-like synoviocytes (RAFLS), mechanisms of Ca -mediated autophagy were analysed by histological, immunohistochemical and flow cytometric techniques. Anti-arthritic effects of celastrol, autophagy induction, and growth rate of synovial fibroblasts in AIA rats were monitored by microCT and immunofluorescence staining. mRNA from joint tissues of AIA rats was isolated for transcriptional analysis of inflammatory genes, using siRNA methods to study calmodulin, calpains, and calcineurin. Celastrol inhibited SERCA to induce autophagy-dependent cytotoxicity in RASFs/RAFLS via Ca /calmodulin-dependent kinase kinase-β-AMP-activated protein kinase-mTOR pathway and repressed arthritis symptoms in AIA rats. BAPTA/AM hampered the in vitro and in vivo effectiveness of celastrol. Inflammatory- and autoimmunity-associated genes down-regulated by celastrol in joint tissues of AIA rat were restored by BAPTA/AM. Knockdown of calmodulin, calpains, and calcineurin in RAFLS confirmed the role of Ca in celastrol-regulated gene expression. Celastrol triggered Ca signalling to induce autophagic cell death in RASFs/RAFLS and ameliorated arthritis in AIA rats mediated by calcium-dependent/-binding proteins facilitating the exploitation of anti-arthritic drugs based on manipulation of Ca signalling.