Heme-oxygenase-1 induction and carbon monoxide-releasing molecule inhibit lipopolysaccharide (LPS)-induced high-mobility group box 1 release in vitro and improve survival of mice in LPS- and cecal ligation and puncture-induced sepsis model in vivo.
We examined our hypothesis that heme-oxygenase-1 (HO-1)-derived carbon monoxide (CO) inhibits the release of high-mobility group box 1 (HMGB1) in RAW264.7 cells activated with lipopolysaccharide (LPS) in vitro and in LPS- or cecal ligation and puncture (CLP)-induced septic mice in vivo, so that HO-1 induction or CO improves survival of sepsis in rodents. We found that pretreatment with HO-1 inducers (hemin, cobalt protoporphyrin IX) or transfection of HO-1 significantly inhibited HMGB1 release, which was blocked by HO-1 small interfering RNA, in cells activated by LPS. Carbon monoxide-releasing molecule 2 (CORM-2) but not bilirubin or deferoxamine inhibited HMGB1 release in LPS-activated macrophages. Oxyhemoglobin reversed the effect of HO-1 inducers on HMGB1 release. Translocation of HMGB1 from nucleus to cytosol was significantly inhibited by HO-1 inducers, CORM-2, or HO-1 transfection. Neutralizing antibodies to tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, interferon-beta, and N(omega)-nitro-L-arginine methyl ester hydrochloride but not N-[2-(cyclohexyloxyl)-4-nitrophenyl]-methane sulfonamide (NS-398) significantly inhibited HMGB1 release in LPS-activated cells. Production of TNF-alpha, IL-1beta, and IFN-beta was significantly reduced by pretreatment of HO-1 inducers, CORM-2, or HO-1 transfection in LPS-activated cells. Plasma levels of HMGB1 in mice challenged with LPS or CLP were significantly reduced by the administration of HO-1 inducers or CORM-2, which was accompanied by either reduction (pretreatment) or no change (delayed administration) of serum TNF-alpha and IL-1beta levels. Regardless of pretreatment or delayed administration, CORM-2 and hemin rescued mice from lethal endotoxemia and sepsis induced by LPS or CLP. Taken together, we concluded that HO-1-derived CO reduces HMGB1 release in LPS-activated cells and LPS- or CLP-induced animal model of sepsis.
Department of Pharmacology, School of Medicine, Gyeongsang National University, Jinju, Republic of Korea.
Konstantin
K
Tsoyi
Tae Yu
TY
Lee
Young Soo
YS
Lee
Hye Jung
HJ
Kim
Han Geuk
HG
Seo
Jae Heun
JH
Lee
Ki Churl
KC
Chang
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000818:Animals; D002248:Carbon Monoxide / Q000502:physiology*; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000502:physiology; D016207:Cytokines / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism; D004195:Disease Models, Animal; D024243:HMGB1 Protein / Q000378:metabolism*; D051547:Heme Oxygenase-1 / Q000502:physiology*; D006427:Hemin / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors*; D008565:Membrane Proteins / Q000502:physiology*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000502:physiology; D009942:Organometallic Compounds / Q000494:pharmacology; D011524:Protoporphyrins / Q000494:pharmacology; D018805:Sepsis / Q000401:mortality / Q000628:therapy*
D016207:Cytokines; D024243:HMGB1 Protein; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D009942:Organometallic Compounds; D011524:Protoporphyrins; C447082:tricarbonyldichlororuthenium (II) dimer; C007095:cobaltiprotoporphyrin; D006427:Hemin; D002248:Carbon Monoxide; D052247:Nitric Oxide Synthase Type II; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2