A novel function of artesunate on inhibiting migration and invasion of fibroblast-like synoviocytes from rheumatoid arthritis patients

• Published in: Arthritis research & therapy Vol. 21; no. 1; p. 153
• Main Authors: Ma, Jian-Da, Jing, Jun, Wang, Jun-Wei, Yan, Tao, Li, Qian-Hua, Mo, Ying-Qian, Zheng, Dong-Hui, Gao, Jin-Long, Nguyen, Ky-Anh, Dai, Lie
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 24.06.2019; BioMed Central; BMC
• Subjects: Adult; Antimalarials - pharmacology; Apoptosis; Artesunate; Artesunate - pharmacology; Arthritis; Arthritis, Rheumatoid - drug therapy; Arthritis, Rheumatoid - metabolism; Arthritis, Rheumatoid - pathology; Biopsy, Needle; Blotting, Western; Cancer therapies; Cell adhesion & migration; Cell Movement - drug effects; Cell Proliferation - drug effects; Cells, Cultured; Drug therapy; Drugs; Enzyme-Linked Immunosorbent Assay; Female; Fibroblast-like synoviocytes; Fibroblasts; Fibroblasts - metabolism; Fibroblasts - pathology; Humans; Inflammation; Invasion; Male; Middle Aged; Migration; Motility; Patient outcomes; Pharmaceuticals; Phosphorylation; Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase - genetics; Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase - metabolism; Rheumatoid arthritis; Rheumatology; Signal Transduction - drug effects; Synoviocytes - drug effects; Synoviocytes - metabolism; Synoviocytes - pathology; Systematic review; Tumor necrosis factor-TNF; Young Adult; China; St Louis Missouri; United States > US; Fibroblast-like synoviocytes; Artesunate; Migration; Rheumatoid arthritis; Invasion
• ISSN: 1478-6362; 1478-6354; 1478-6362
• DOI: 10.1186/s13075-019-1935-6

Anti-malarial drug artesunate can suppress inflammation and prevent cartilage and bone destruction in collagen-induced arthritis model in rats-suggesting it may be a potent drug for rheumatoid arthritis (RA) therapy. We aimed to investigate its effect on the invasive property of fibroblast-like synoviocytes (FLS) from patients with RA. Synovial tissues were obtained by closed needle biopsy from active RA patients, and FLS were isolated and cultured in vitro. RA-FLS were treated with artesunate at various concentrations, while methotrexate or hydroxychloroquine was employed as comparator drugs. Cell viability, proliferation, cell cycle, apoptosis, migration, invasion, and pseudopodium formation of RA-FLS were assessed by CCK-8 assays, EdU staining, Annexin V-FITC/PI staining, transwell assays, or F-actin staining, respectively. Further, relative changes of expressed proteases were analyzed by Proteome profiler human protease array and verified by quantitative real-time PCR (qPCR), Western blot, and ELISA. The expression of signaling molecules of MAPK, NF-κB, AP-1, and PI3K/Akt pathways were measured by qPCR and Western blot. PDK-1 knockdown by specific inhibitor AR-12 or siRNA transfection was used to verify the pharmacological mechanism of artesunate on RA-FLS. Artesunate significantly inhibited the migration and invasion of RA-FLS in a dose-dependent manner with or without TNF-α stimulation. The effect was mediated through artesunate inhibition of MMP-2 and MMP-9 production, and pre-treatment with exogenous MMP-9 reversed the inhibitory effect of artesunate on RA-FLS invasion. Artesunate had a stronger inhibitory effect on migration and invasion of RA-FLS as well as greater anti-inflammatory effect than those of hydroxychloroquine. Similar inhibitory effect was detected between artesunate and methotrexate, and synergy was observed when combined. Mechanistically, artesunate significantly inhibited PDK-1 expression as well as Akt and RSK2 phosphorylation-in a similar manner to PDK-1-specific inhibitor AR-12 or PDK-1 knockdown by siRNA transfection. This inhibition results in suppression of RA-FLS migration and invasion as well as decreased MMP-2 and MMP-9 expression. Our study demonstrates artesunate is capable of inhibiting migration and invasion of RA-FLS through suppression of PDK1-induced activation of Akt and RSK2 phosphorylation-suggesting that artesunate may be a potential disease-modifying anti-rheumatic drug for RA.