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Abstract Number: 963

Artesunate Inhabits Migration and Invasion of Fibroblast-like Synoviocytes and Matrix Metalloproteinases Expression Via Suppression of PI3K/Akt Pathway in Rheumatoid Arthritis

Jian-Da Ma1, Jun Jing1, Tao Yan2, Ying-Qian Mo1 and Lie Dai1, 1Department of Rheumatology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China, 2Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China

Meeting: 2017 ACR/ARHP Annual Meeting

Date of first publication: September 18, 2017

Keywords: Fibroblasts, matrix metalloproteinase (MMP) and rheumatoid arthritis (RA)

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Session Information

Date: Monday, November 6, 2017

Title: Cytokines, Mediators, Cell-Cell Adhesion, Cell Trafficking and Angiogenesis Poster II: Mesenchymal Cells Do React - But How?

Session Type: ACR Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose: Fibroblast-like synoviocytes (FLS) play major roles on joint destruction in rheumatoid arthritis (RA) through migrating and invasing cartilage and bone by secreting proteases such as matrix metalloproteinases (MMPs).Recent studies show that artesunate, an important artemisinin derivative, may inhibit proinflammatory cytokines secretion of RA-FLS. We aim to investigate the effect of artesunate on migration and invasion of RA-FLS and itsunderlying mechanism

Methods: FLS isolated from knee synovium of active RA patients were cultured in vitro. The effects of artesunate, methotrexate (MTX) or hydroxychloroquine (HCQ) on cell viability and anti-proliferation were measured by CCK-8 assay. Effects on migration and invasion capacity were detected by wound healing and transwell assays. Differential expression of MMPs were analyzed by Proteome profiler human protease array (R&D Systems) and furtherly verifiedby quantitative real-time PCR, western blot (WB) and ELISA. The expression of PI3K, PIP2, PIP3, PDK1 and Akt in PI3K/Akt signal pathway was measured by WB.

Results: The IC50 value of artesunate, MTX and HCQ on RA-FLS were 6891μM, 181.4nM and 5433μM respectively. Compared with untreated group, IC1-20μM, IC3-40μM, IC5-60μM of artesunate, IC5-10nM of MTX and IC5-20μM of HCQ showed no significant change in proliferation even after 72h. Wound healing and migration assays for 12h and invasion assay for 24h showed artesunate inhabits the migration and invasion of RA-FLS in a dose-dependent manner. MTX also has inhibition effect on the migration and invasion of RA-FLS, but HCQ not (Fig. A). Proteome profiler human protease array of culture supernatant showed that 60μM artesunate markedly inhibited MMP-2/9 expression (Fig. B). Further verification showed that RA-FLS pretreated with 60μM artesunate attenuated MMP-2/9 mRNA and protein expression, and ELISA results showed 60μM artesunate decreased MMP-2 and MMP-9 concentration of 5.82±0.45 and 11.74±1.30ng/mL respectively (Fig. C). Further pre-treatment with recombination human MMP-2 (6 ng/ml) or MMP-9 (12 ng/ml) for 24h could reverse inhibitory effect of 60μM artesunate on invasion, but not migration (Fig. D). Quantitative real-time PCR and WB analysis showed that PDK1 expression and Akt activity (phophso-Akt/Akt) in 60μM artesunate treatment group was significantly lower than that in untreated group which indicated that artesunate suppressed generation of PDK1-induced activation of Akt (Fig. E).

Conclusion: Artesunate could inhibit migration and invasion of RA-FLS and MMP-2/9 expression through suppressing PDK1-induced activation of Akt.


Disclosure: J. D. Ma, National Natural Science Foundation of China (no. 81471597 and 81671612), 2,Guangdong Natural Science Foundation (no.2014A030313074), 2,Scientific Program of Traditional Chinese Medicine Bureau of Guangdong Province (no. 20161058), 2,Medical Science Research Grant of Guangdong Province, China (no. A2017109), 2; J. Jing, National Natural Science Foundation of China (no. 81471597 and 81671612), 2,Guangdong Natural Science Foundation (no.2014A030313074), 2,Scientific Program of Traditional Chinese Medicine Bureau of Guangdong Province (no. 20161058), 2; T. Yan, National Natural Science Foundation of China (no. 81471597 and 81671612), 2,Guangdong Natural Science Foundation (no.2014A030313074), 2,Scientific Program of Traditional Chinese Medicine Bureau of Guangdong Province (no. 20161058), 2; Y. Q. Mo, National Natural Science Foundation of China (no. 81471597 and 81671612), 2,Guangdong Natural Science Foundation (no.2014A030313074), 2,Scientific Program of Traditional Chinese Medicine Bureau of Guangdong Province (no. 20161058), 2; L. Dai, National Natural Science Foundation of China (no. 81471597 and 81671612), 2,Guangdong Natural Science Foundation (no.2014A030313074), 2,Scientific Program of Traditional Chinese Medicine Bureau of Guangdong Province (no. 20161058), 2.

To cite this abstract in AMA style:

Ma JD, Jing J, Yan T, Mo YQ, Dai L. Artesunate Inhabits Migration and Invasion of Fibroblast-like Synoviocytes and Matrix Metalloproteinases Expression Via Suppression of PI3K/Akt Pathway in Rheumatoid Arthritis [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/artesunate-inhabits-migration-and-invasion-of-fibroblast-like-synoviocytes-and-matrix-metalloproteinases-expression-via-suppression-of-pi3kakt-pathway-in-rheumatoid-arthritis/. Accessed .
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