title: Carbon monoxide-releasing molecule-3 suppresses Prevotella intermedia lipopolysaccharide-induced production of nitric oxide and interleukin-1β in murine macrophages.
abstract: This study was performed to analyze the effect of carbon monoxide (CO)-releasing molecule-3 (CORM-3) in alleviating the production of proinflammatory mediators in macrophages treated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen associated with periodontal disease, and its possible mechanisms of action. LPS was isolated using the hot phenol-water method. Culture supernatants were assayed for nitric oxide (NO) and interleukin-1β (IL-1β). Gene expression was quantified by real-time PCR, and protein expression by immunoblotting. DNA-binding activities of NF-κB subunits were determined using an ELISA-based kit. CORM-3 suppressed the production of inducible NO synthase (iNOS)-derived NO and IL-1β at both gene transcription and translation levels in P. intermedia LPS-activated RAW264.7 cells. CORM-3 enhanced heme oxygenase-1 (HO-1) expression in cells stimulated with P. intermedia LPS, and inhibition of HO-1 activity by SnPP notably reversed the suppressive effect of CORM-3 on LPS-induced production of NO. LPS-induced phosphorylation of p38 and JNK was not affected by CORM-3. CORM-3 did not influence P. intermedia LPS-induced degradation of IκB-α. Instead, nuclear translocation of NF-κB p65 and p50 subunits was blocked by CORM-3 in LPS-treated cells. In addition, CORM-3 reduced LPS-induced p65 and p50 binding to DNA. Besides, CORM-3 significantly suppressed P. intermedia LPS-induced phosphorylation of STAT1. Overall, this study indicates that CORM-3 suppresses the production of NO and IL-1β in P. intermedia LPS-activated murine macrophages via HO-1 induction and inhibition of NF-κB and STAT1 pathways. The modulation of host inflammatory response by CORM-3 would be an attractive therapeutic approach to attenuate the progression of periodontal disease.
authors.affiliation: Department of Biological Science, College of Medical and Life Sciences, Silla University, Busan, South Korea.
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Choi
authors.affiliation: Department of Biological Science, College of Medical and Life Sciences, Silla University, Busan, South Korea.
authors.forename: So-Hui
authors.initials: SH
authors.lastname: Choe
authors.affiliation: Department of Biological Science, College of Medical and Life Sciences, Silla University, Busan, South Korea.
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Hyeon
authors.affiliation: Department of Periodontology, School of Dentistry, Pusan National University, 49 Busandaehak-ro, Mulgeum-eup, Yangsan, Gyeongsangnam-do, South Korea; Dental Research Institute, Pusan National University Dental Hospital, Yangsan, Gyeongsangnam-do, South Korea.
authors.forename: Jeom-Il
authors.initials: JI
authors.lastname: Choi
authors.affiliation: Department of Biological Science, College of Medical and Life Sciences, Silla University, Busan, South Korea.
authors.forename: In Soon
authors.initials: IS
authors.lastname: Choi
authors.affiliation: Department of Periodontology, School of Dentistry, Pusan National University, 49 Busandaehak-ro, Mulgeum-eup, Yangsan, Gyeongsangnam-do, South Korea; Dental Research Institute, Pusan National University Dental Hospital, Yangsan, Gyeongsangnam-do, South Korea; Institute of Translational Dental Sciences, Pusan National University, Yangsan, Gyeongsangnam-do, South Korea. Electronic address: sungjokim@pusan.ac.kr.
authors.forename: Sung-Jo
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D002248:Carbon Monoxide / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D051547:Heme Oxygenase-1 / Q000096:biosynthesis; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000096:biosynthesis*; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D009942:Organometallic Compounds / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D018713:Prevotella intermedia / Q000737:chemistry*; D000067996:RAW 264.7 Cells; D050794:STAT1 Transcription Factor / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D018836:Inflammation Mediators; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D016328:NF-kappa B; D009942:Organometallic Compounds; D050794:STAT1 Transcription Factor; C476530:tricarbonylchloro(glycinato)ruthenium(II); D009569:Nitric Oxide; D002248:Carbon Monoxide; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases
keywords: CORM-3; HO-1; Lipopolysaccharide; NF-κB; Proinflammatory mediators; STAT1