title: Anti-inflammatory effects of ononin on lipopolysaccharide-stimulated RAW 264.7 cells.
abstract: Increasing evidence has shown that ononin, a major isoflavone, has anti-inflammatory effects on lipopolysaccharide (LPS)-induced inflammation. However, the molecular mechanisms underlying the anti-inflammatory effects of ononin are still unclear. In the present study, we investigated these effects and the underlying mechanisms of ononin on LPS-induced inflammatory responses. Mouse RAW 264.7 cells were treated with 1μg/mL LPS and 5, 25, 50, 100 or 150μM ononin for 18h. Cell viability was assessed using MTT assays, and the production of nitric oxide (NO), prostaglandin E2(PGE2) and the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 in cultures was examined by Griess and ELISA analyses. qRT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2). Mitogen-activated protein kinases (MAPKs) and nuclear transcription factor Kappa-B (NF-κB) signalling pathway-related proteins were assessed by western blot assays. The results showed that cell viability was not significantly affected by up to 100μM ononin. The production of NO, PGE2and the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 in the cultures, the mRNA expression of two major inflammatory mediators, COX-2 and iNOS, and the expression of phosphorylated IκB-α, ERK, JNK, and p38 MAPKs proteins in LPS-treated cells were significantly increased. These changes could be reversed by treatment with ononin in a concentration-dependent manner (P<0.05). The results suggest that ononin has anti-inflammatory effects on LPS-induced inflammatory responses by inhibiting the NF-κB and MAPK pathways and may be a potential treatment for inflammation.
authors.affiliation: School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China; School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Dong
authors.affiliation: School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yin
authors.affiliation: School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
authors.forename: Yuanbin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Ningxia Medical University, Yinchuan 750004, China.
authors.forename: Xueyan
authors.initials: X
authors.lastname: Fu
authors.affiliation: School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang 110016, China. Electronic address: jincailu@126.com.
authors.forename: Jincai
authors.initials: J
authors.lastname: Lu
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005960:Glucosides / Q000494:pharmacology*; D007249:Inflammation / Q000276:immunology*; D007529:Isoflavones / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000276:immunology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000276:immunology; D051379:Mice; D016328:NF-kappa B / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D000067996:RAW 264.7 Cells; D015398:Signal Transduction / Q000187:drug effects* / Q000276:immunology
chemical_list: D000893:Anti-Inflammatory Agents; D005960:Glucosides; D007529:Isoflavones; D008070:Lipopolysaccharides; D016328:NF-kappa B; C526426:calycosin-7-O-beta-D-glucoside
keywords: Inflammation; Inflammatory cytokines; Mapk; NF-κB; Ononin