Abstract
The aim of this study was to investigate the protective effects of cepharanthine (CEP) on inflammation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells in vitro and a LPS-induced lung injury model in vivo. RAW264.7 cells were treated with various concentrations of CEP for 1 h followed by incubation with or without 1 μg/ml LPS for 18 h. TNF-α, IL-6, and IL-1β in the supernatants were measured by ELISA. Nuclear factor-κB (NF-κB) and mitogen-activated protein kinase pathways were analyzed by Western blot. Mice were randomly divided into control group, LPS group, CEP + LPS group, and dexamethasone + LPS group. A male BALB/c mouse model of acute lung injury was induced by LPS. Bronchoalveolar lavage fluid was collected for inflammatory cell count and cytokine assays. Histopathologic examination was performed on mice that were not subjected to bronchoalveolar lavage fluid collection. CEP dose-dependently inhibited the release of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. Significantly, CEP dose-dependently suppressed NF-κB activation, IκBα degradation, and phosphorylation of ERK, JNK, and p38 induced by LPS. In vivo, it was also observed that CEP attenuated lung histopathologic changes and down-regulated the level of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, in the mouse acute lung injury model. These results suggest that CEP potentially decreases inflammation in vitro and in vivo and might be a therapeutic agent against inflammatory diseases.
Similar content being viewed by others
References
Rubenfeld, G.D. 2003. Epidemiology of acute lung injury. Critical Care Medicine 31: S276–S284.
Zhang, X., K. Song, H. Xiong, H. Li, X. Chu, and X. Deng. 2009. Protective effect of florfenicol on acute lung injury induced by lipopolysaccharide in mice. International Immunopharmacology 9: 1525–1529.
Atabai, K., and M.A. Matthay. 2002. The pulmonary physician in critical care. 5: Acute lung injury and the acute respiratory distress syndrome: definitions and epidemiology. Thorax 57: 452–458.
Rubenfeld, G.D., E. Caldwell, E. Peabody, J. Weaver, D.P. Martin, M. Neff, E.J. Stern, and L.D. Hudson. 2005. Incidence and outcomes of acute lung injury. New England Journal of Medicine 353: 1685–1693.
Boje, K.M. 2004. Nitric oxide neurotoxicity in neurodegenerative diseases. Frontiers in Bioscience 9: 763–776.
Takeuchi, H., S. Jin, J. Wang, G. Zhang, J. Kawanokuchi, R. Kuno, Y. Sonobe, T. Mizuno, and A. Suzumura. 2006. Tumor necrosis factor-alpha induces neurotoxicity via glutamate release from hemichannels of activated microglia in an autocrine manner. Journal of Biological Chemistry 281: 21362–21368.
Hartlage-Rubsamen, M., R. Lemke, and R. Schliebs. 1999. Interleukin-1beta, inducible nitric oxide synthase, and nuclear factor-kappaB are induced in morphologically distinct microglia after rat hippocampal lipopolysaccharide/interferon-gamma injection. Journal of Neuroscience Research 57: 388–398.
Kang, J., Y. Zhang, X. Cao, J. Fan, G. Li, Q. Wang, Y. Diao, Z. Zhao, L. Luo, and Z. Yin. 2012. Lycorine inhibits lipopolysaccharide-induced iNOS and COX-2 up-regulation in RAW264.7 cells through suppressing P38 and STATs activation and increases the survival rate of mice after LPS challenge. International Immunopharmacology 12: 249–256.
Medzhitov, R., and J.C. Kagan. 2006. Phosphoinositide-mediated adaptor recruitment controls toll-like receptor signaling. Cell 125: 943–955.
Jung, H.W., R. Mahesh, J.H. Park, Y.C. Boo, K.M. Park, and Y.K. Park. 2010. Bisabolangelone isolated from Ostericum koreanum inhibits the production of inflammatory mediators by down-regulation of NF-kappaB and ERK MAP kinase activity in LPS-stimulated RAW264.7 cells. International Immunopharmacology 10: 155–162.
Qiu, X., H. Li, H. Tang, Y. Jin, W. Li, YuSun, PingFeng, X. Sun, and Z. Xia. 2011. Hydrogen inhalation ameliorates lipopolysaccharide-induced acute lung injury in mice. International Immunopharmacology 11: 2130–2137.
Wang, D., M. Liu, Y. Wang, M. Luo, J. Wang, C. Dai, P. Yan, X. Zhang, C. Tang, and J. Xiao. 2011. Synthetic LXR agonist T0901317 attenuates lipopolysaccharide-induced acute lung injury in rats. International Immunopharmacology 11: 2098–2103.
Ware, L.B., and M.A. Matthay. 2000. The acute respiratory distress syndrome. New England Journal of Medicine 342: 1334–1349.
Lee, W.L., and G.P. Downey. 2001. Neutrophil activation and acute lung injury. Current Opinion in Critical Care 7: 1–7.
Kudo, K., S. Hagiwara, A. Hasegawa, J. Kusaka, H. Koga, and T. Noguchi. 2011. Cepharanthine exerts anti-inflammatory effects via NF-kappaB inhibition in a LPS-induced rat model of systemic inflammation. Journal of Surgical Research 171: 199–204.
Murakami, K., K. Okajima, and M. Uchiba. 2000. The prevention of lipopolysaccharide-induced pulmonary vascular injury by pretreatment with cepharanthine in rats. American Journal of Respiratory and Critical Care Medicine 161: 57–63.
Choi, Y.H., G.Y. Jin, G.Z. Li, and G.H. Yan. 2011. Cornuside suppresses lipopolysaccharide-induced inflammatory mediators by inhibiting nuclear factor-kappa B activation in RAW 264.7 macrophages. Biological and Pharmaceutical Bulletin 34: 959–966.
Nys, M., J.C. Preiser, G. Deby-Dupont, Y. Habraken, M. Mathy-Hartert, P. Damas, and M. Lamy. 2005. Nitric oxide-related products and myeloperoxidase in bronchoalveolar lavage fluids from patients with ALI activate NF-kappa B in alveolar cells and monocytes. Vascular Pharmacology 43: 425–433.
Satoh, H., G.S. Firestein, P.B. Billings, J.P. Harris, and E.M. Keithley. 2003. Proinflammatory cytokine expression in the endolymphatic sac during inner ear inflammation. Journal of the Association for Research in Otolaryngology 4: 139–147.
Libby, P. 2000. Coronary artery injury and the biology of atherosclerosis: inflammation, thrombosis, and stabilization. American Journal of Cardiology 86: 3J–8J. discussion 8J-9J.
Gilmore, T.D. 2006. Introduction to NF-kappaB: players, pathways, perspectives. Oncogene 25: 6680–6684.
Rao, K.M. 2001. MAP kinase activation in macrophages. Journal of Leukocyte Biology 69: 3–10.
Jiang, J.X., Y. Zhang, S.H. Ji, P. Zhu, and Z.G. Wang. 2002. Kinetics of mitogen-activated protein kinase family in lipopolysaccharide-stimulated mouse Kupffer cells and their role in cytokine production. Shock 18: 336–341.
Deng, X.M., X.X. Ci, R. Ren, K. Xu, H.Y. Li, Q.L. Yu, Y. Song, D.C. Wang, and R.T. Li. 2010. Schisantherin A exhibits anti-inflammatory properties by down-regulating NF-kappaB and MAPK signaling pathways in lipopolysaccharide-treated RAW 264.7 cells. Inflammation 33: 126–136.
Lai, C.S., Y.S. Lai, D.H. Kuo, C.H. Wu, C.T. Ho, and M.H. Pan. 2011. Magnolol potently suppressed lipopolysaccharide-induced iNOS and COX-2 expression via downregulating MAPK and NF-kappa B signaling pathways. Journal of Functional Foods 3: 198–206.
Bhatia, M., and S. Moochhala. 2004. Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome. Journal of Pathology 202: 145–156.
Donnelly, S.C., C. Haslett, P.T. Reid, I.S. Grant, W.A. Wallace, C.N. Metz, L.J. Bruce, and R. Bucala. 1997. Regulatory role for macrophage migration inhibitory factor in acute respiratory distress syndrome. Nature Medicine 3: 320–323.
Giebelen, I.A., D.J. van Westerloo, G.J. LaRosa, A.F. de Vos, and T. van der Poll. 2007. Local stimulation of alpha7 cholinergic receptors inhibits LPS-induced TNF-alpha release in the mouse lung. Shock 28: 700–703.
Christman, J.W., R.T. Sadikot, and T.S. Blackwell. 2000. The role of nuclear factor-kappa B in pulmonary diseases. Chest 117: 1482–1487.
McCoy, M.K., K.A. Ruhn, A. Blesch, and M.G. Tansey. 2011. TNF: a key neuroinflammatory mediator of neurotoxicity and neurodegeneration in models of Parkinson's disease. Advances in Experimental Medicine and Biology 691: 539–540.
Martin, T.R. 1999. Lung cytokines and ARDS: Roger S. Mitchell Lecture. Chest 116: 2S–8S.
Hodge, D.R., E.M. Hurt, and W.L. Farrar. 2005. The role of IL-6 and STAT3 in inflammation and cancer. European Journal of Cancer 41: 2502–2512.
Minamino, T., and I. Komuro. 2006. Regeneration of the endothelium as a novel therapeutic strategy for acute lung injury. Journal of Clinical Investigation 116: 2316–2319.
Meduri, G.U. 2002. Clinical review: a paradigm shift: the bidirectional effect of inflammation on bacterial growth. Clinical implications for patients with acute respiratory distress syndrome. Critical Care 6: 24–29.
Sun, J., D. Yang, S. Li, Z. Xu, X. Wang, and C. Bai. 2009. Effects of curcumin or dexamethasone on lung ischaemia–reperfusion injury in rats. European Respiratory Journal 33: 398–404.
Acknowledgments
This work was supported by a grant from the National Natural Science Foundation of China (nos. 31272566 and 31272541).
Conflict of interest
The authors declare that there are no conflicts of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Huang, H., Hu, G., Wang, C. et al. Cepharanthine, an Alkaloid from Stephania cepharantha Hayata, Inhibits the Inflammatory Response in the RAW264.7 Cell and Mouse Models. Inflammation 37, 235–246 (2014). https://doi.org/10.1007/s10753-013-9734-8
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10753-013-9734-8