title: Casticin inhibits COX-2 and iNOS expression via suppression of NF-κB and MAPK signaling in lipopolysaccharide-stimulated mouse macrophages.
abstract: BACKGROUND
  The fruits of Vitex rotundifolia L. are widely used to treat inflammation of the airway in Traditional Chinese medicine. Previous studies found that casticin, isolated from Vitex rotundifolia, could induce apoptosis of tumor cells. In this study, we evaluated the anti-inflammatory effects of casticin and its underlying molecular mechanism in lipopolysaccharide (LPS)-stimulated macrophages.
  
  
  METHODS
  RAW264.7 cells were pretreated with various concentrations of casticin (0.3-10μM), and then treated with LPS to induce inflammation. We assayed the levels of proinflammatory cytokines and prostaglandin E2 (PGE2) using ELISA, and examined the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, and heme oxygenase (HO)-1 by Western blot. We also investigated the anti-inflammatory molecular mechanism by analyzing inflammatory-associated signaling pathways, including the nuclear transcription factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways.
  
  
  RESULTS
  We found casticin inhibited the levels of nitric oxide and PGE2, and decreased the production of proinflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor α (TNF-α). In addition, iNOS and COX-2 expression levels were suppressed and casticin increased HO-1 and Nrf2 production in a concentration-dependent manner. Furthermore, casticin significantly inhibited NF-κB subunit p65 proteins in the nucleus and decreased Akt and MAPK activation.
  
  
  CONCLUSIONS
  These results suggest that the anti-inflammatory effect of casticin is due to inhibition of proinflammatory cytokines and mediators by blocking the NF-κB, Akt, and MAPK signaling pathways.
authors.affiliation: Department of Nursing, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.
authors.forename: Chian-Jiun
authors.initials: CJ
authors.lastname: Liou
authors.affiliation: Department of Nursing, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Len
authors.affiliation: Department of Nutrition and Health Sciences, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.
authors.forename: Shu-Ju
authors.initials: SJ
authors.lastname: Wu
authors.affiliation: Department of Cosmetic Sciences, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.
authors.forename: Chwan-Fwu
authors.initials: CF
authors.lastname: Lin
authors.affiliation: Department of Nutrition and Health Sciences, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC.
authors.forename: Xin-Ling
authors.initials: XL
authors.lastname: Wu
authors.affiliation: Department of Nutrition and Health Sciences, Chang Gung University of Science and Technology, 261 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC; Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC; Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kwei-Shan, Tao-Yuan 333, Taiwan, ROC. Electronic address: wchuang@gw.cgust.edu.tw.
authors.forename: Wen-Chung
authors.initials: WC
authors.lastname: Huang
mesh_terms: D000818:Animals; D002460:Cell Line; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism
chemical_list: D005419:Flavonoids; D008070:Lipopolysaccharides; D016328:NF-kappa B; C054133:casticin; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D051057:Proto-Oncogene Proteins c-akt
keywords: Casticin; Inducible nitric oxide synthase; MAPK; NF-κB; Proinflammatory cytokines