Inhibitory Effects of Polydatin on Lipopolysaccharide-Stimulated RAW 264.7 Cells
The purpose of this study was to evaluate the effects of polydatin (PD) on cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expressions at protein and transcriptional levels, as well as the production of prostaglandin E2 (PGE2) and nitric oxide (NO) in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells. To elucidate the underlying mechanism responsible for these symptoms, we investigated the phosphorylation of mitogen-activated protein kinase (MAPK) pathway and nuclear factor-κB (NF-κB) expression. NO was analyzed with the Griess method. PGE2 was measured by enzyme-linked immunosorbent assay (ELISA). iNOS and COX-2 messenger RNA (mRNA) were identified by qPCR assay. iNOS, COX-2, NF-κB, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 protein expressions were detected with Western blot. The results revealed that PD effectively inhibited NO and PGE2 production, and it is not surprising that PD reduced iNOS and COX-2 expression at protein and transcriptional levels. Additionally, PD significantly ameliorated the activation of NF-κB and the phosphorylation of MAPKs in LPS-induced RAW 264.7 macrophages. These findings suggested that PD exerted potent anti-inflammatory activity in macrophages.
KEY WORDSpolydatin LPS RAW 264.7 cells
- 2.Salvemini, D., H. Ischiropoulos, and S. Cuzzocrea. 2003. Roles of nitric oxide and superoxide in inflammation. Methods in Molecular Biology (Clifton, NJ) 225: 291–303.Google Scholar
- 3.Wang, J., Y.T. Liu, L. Xiao, L. Zhu, Q. Wang, T. Yan. 2014. Anti-inflammatory effects of apigenin in lipopolysaccharide-induced inflammatory in acute lung injury by suppressing COX-2 and NF-kB Pathway. Inflammation. ;37:2085–2090. doi: 10.1007/s10753-014-9942-x.
- 4.Mancini, A.D., and J.A. Di Battista. 2011. The cardinal role of the phospholipase A(2)/cyclooxygenase-2/prostaglandin E synthase/prostaglandin E(2) (PCPP) axis in inflammostasis. Inflammation Research: Official Journal of the European Histamine Research Society [Et al.] 60: 1083–1092.CrossRefGoogle Scholar
- 8.Van Laecke, S., W. Van Biesen, R. Vanholder. 2014. The paradox of bardoxolone methyl: A call for every witness on the stand? Diabetes, Obesity & Metabolism. 17:9–14.Google Scholar
- 9.Park, K.I., S.R. Kang, H.S. Park, H. do Lee, A. Nagappan, J.A. Kim, et al. 2012. Regulation of proinflammatory mediators via NF-kappaB and p38 MAPK-dependent mechanisms in RAW 264.7 macrophages by polyphenol components isolated from Korea Lonicera japonica THUNB. Evidence-Based Complementary and Alternative Medicine: eCAM 2012: 828521.PubMedGoogle Scholar
- 17.Ignarro, L.J., G.M. Buga, K.S. Wood, R.E. Byrns, and G. Chaudhuri. 1987. Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proceedings of the National Academy of Sciences of the United States of America 84: 9265–9269.CrossRefPubMedCentralPubMedGoogle Scholar
- 19.Asano, K., C.B. Chee, B. Gaston, C.M. Lilly, C. Gerard, J.M. Drazen, et al. 1994. Constitutive and inducible nitric oxide synthase gene expression, regulation, and activity in human lung epithelial cells. Proceedings of the National Academy of Sciences of the United States of America 91: 10089–10093.CrossRefPubMedCentralPubMedGoogle Scholar
- 23.Geiser, T., K. Atabai, P.H. Jarreau, L.B. Ware, J. Pugin, and M.A. Matthay. 2001. Pulmonary edema fluid from patients with acute lung injury augments in vitro alveolar epithelial repair by an IL-1beta-dependent mechanism. American Journal of Respiratory and Critical Care Medicine 163: 1384–1388.CrossRefPubMedGoogle Scholar
- 24.Cheng, B.C., X.Q. Ma, H.Y. Kwan, K.W. Tse, H.H. Cao, T. Su, et al. 2014. A herbal formula consisting of Rosae Multiflorae Fructus and Lonicerae Japonicae Flos inhibits inflammatory mediators in LPS-stimulated RAW 264.7 macrophages. Journal of Ethnopharmacology 153: 922–927.CrossRefPubMedGoogle Scholar
- 25.Miggin, S.M., E. Palsson-McDermott, A. Dunne, C. Jefferies, E. Pinteaux, K. Banahan, et al. 2007. NF-kappaB activation by the Toll-IL-1 receptor domain protein MyD88 adapter-like is regulated by caspase-1. Proceedings of the National Academy of Sciences of the United States of America 104: 3372–3377.CrossRefPubMedCentralPubMedGoogle Scholar
- 28.Wu, D., S. Zheng, W. Li, L. Yang, Y. Liu, X. Zheng, et al. 2013. Novel biphasic role of resolvin D1 on expression of cyclooxygenase-2 in lipopolysaccharide-stimulated lung fibroblasts is partly through PI3K/AKT and ERK2 pathways. Mediators of Inflammation 2013: 964012.PubMedCentralPubMedGoogle Scholar
- 31.Yun, K.J., J.Y. Kim, J.B. Kim, K.W. Lee, S.Y. Jeong, H.J. Park, et al. 2008. Inhibition of LPS-induced NO and PGE2 production by asiatic acid via NF-kappa B inactivation in RAW 264.7 macrophages: Possible involvement of the IKK and MAPK pathways. International Immunopharmacology 8: 431–441.CrossRefPubMedGoogle Scholar