Rebamipide Suppresses Monosodium Urate Crystal-Induced Interleukin-1β Production Through Regulation of Oxidative Stress and Caspase-1 in THP-1 Cells
- 685 Downloads
This study investigated the effect of rebamipide on activation of the NLRP3 inflammasome and generation of reactive oxygen species (ROS) in monosodium urate (MSU) crystal-induced interleukin-1β (IL-1β) production. Human monocyte cell line THP-1 and human umbilical venous endothelial cells (HUVECs) were used to assess the inflammatory response to MSU crystals. NADP/NADPH activity assays were used as a marker of ROS generation. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were performed to evaluate levels of IL-1β, caspase-1, NLRP3, associated speck-like protein (ASC), nuclear factor-κB (NF-κB), p65, IκBα, intercellular adhesion molecule 1 (ICAM-1), and vascular cell adhesion molecule 1 (VCAM-1). Experimental pharmaceuticals included rebamipide, colchicine, dexamethasone, and ascorbic acid. In THP-1 cells, treatment with MSU crystals increased NADP/NADPH ratios and IL-1β expression, and both of these responses were potently inhibited by addition of rebamipide. Rebamipide also attenuated enhanced expression of caspase-1 gene by MSU crystals (p < 0.05). Western blotting demonstrated that MSU crystals stimulated caspase-1 but not NLRP3 and ASC activation. Similarly, MSU crystals activated the NF-κB pathway, which in turn was blocked by rebamipide. Stimulation of HUVECs with MSU crystals increased expression of VCAM-1 and ICAM-1, which were markedly inhibited by both rebamipide and dexamethasone. This study demonstrated that rebamipide inhibits IL-1β activation through suppression of ROS-mediated NF-κB signaling pathways and caspase-1 activation in MSU crystal-induced inflammation.
KEY WORDSuric acid interleukin-1β inflammasome inflammation rebamipide
This work was supported by a grant from the Research Institute of Medical Science, Catholic University of Daegu (2015).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflicts of interest concerning this article.
- 9.Farhadi, A., A. Keshavarzian, L.R. Fitzpatrick, E. Mutlu, Y. Zhang, and A. Banan. 2002. Modulatory effects of plasma and colonic milieu of patients with ulcerative colitis on neutrophil reactive oxygen species production in presence of a novel antioxidant, rebamipide. Digestive Diseases and Sciences 47: 1342–1348.CrossRefPubMedGoogle Scholar
- 12.Moon, S.J., Y.J. Woo, J.H. Woo, M.K. Park, H.J. Oh, J.S. Park, E.K. Kim, M.L. Cho, S.H. Park, H.Y. Kim, and J.K. Min. 2012. Rebamipide attenuates pain severity and cartilage degeneration in a rat model of osteoarthritis by downregulating oxidative damage and catabolic activity in chondrocytes. Osteoarthritis and Cartilage 20: 1426–1438.CrossRefPubMedGoogle Scholar
- 14.Sugai, S., H. Takahashi, S. Ohta, M. Nishinarita, M. Takei, S. Sawada, K. Yamaji, H. Oka, H. Umehara, I. Koni, E. Sugiyama, S. Nishiyama, and A. Kawakami. 2009. Efficacy and safety of rebamipide for the treatment of dry mouth symptoms in patients with Sjögren’s syndrome: a double-blind placebo-controlled multicenter trial. Modern Rheumatology 19: 114–124.CrossRefPubMedGoogle Scholar
- 19.Sautin, Y.Y., T. Nakagawa, S. Zharikov, and R.J. Johnson. 2007. Adverse effects of the classic antioxidant uric acid in adipocytes: NADPH oxidase-mediated oxidative/nitrosative stress. American Journal of Respiratory and Critical Care Medicine 293: C584–C596.Google Scholar
- 20.Kostura, M.J., M.J. Tocci, G. Limjuco, J. Chin, P. Cameron, A.G. Hillman, N.A. Chartrain, and J.A. Schmidt. 1989. Identification of a monocyte specific pre-interleukin 1 beta convertase activity. Proceedings of the National Academy of Sciences of the United States of America 86: 5227–5231.PubMedCentralCrossRefPubMedGoogle Scholar
- 25.Hiscott, J., J. Marois, J. Garoufalis, M. D'Addario, A. Roulston, I. Kwan, N. Pepin, J. Lacoste, H. Nguyen, G. Bensi, and M. Fenton. 1993. Characterization of a functional NF-κB site in the human interleukin 1β promoter: evidence for a positive autoregulatory loop. Molecular and Cellular Biology 13: 6231–6240.PubMedCentralCrossRefPubMedGoogle Scholar