Tsoyi Konstantin, Lee Tae Yu, Lee Young Soo, Kim Hye Jung, Seo Han Geuk, Lee Jae Heun, Chang Ki Churl
Department of Pharmacology, School of Medicine, Gyeongsang National University, Jinju, Republic of Korea.
Mol Pharmacol. 2009 Jul;76(1):173-82. doi: 10.1124/mol.109.055137. Epub 2009 Apr 14.
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.
血红素加氧酶-1（HO-1）衍生的一氧化碳（CO）在体外可抑制脂多糖（LPS）激活的RAW264.7细胞中高迁移率族蛋白B1（HMGB1）的释放，在体内可抑制LPS或盲肠结扎穿孔（CLP）诱导的脓毒症小鼠中HMGB1的释放，从而HO-1的诱导或CO可提高啮齿动物脓毒症的存活率。我们发现，用HO-1诱导剂（血红素、钴原卟啉IX）预处理或HO-1转染可显著抑制LPS激活的细胞中HMGB1的释放，而HO-1小干扰RNA可阻断这种抑制作用。一氧化碳释放分子2（CORM-2）而非胆红素或去铁胺可抑制LPS激活的巨噬细胞中HMGB1的释放。氧合血红蛋白可逆转HO-1诱导剂对HMGB1释放的作用。HO-1诱导剂、CORM-2或HO-1转染可显著抑制HMGB1从细胞核向细胞质的转位。肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、干扰素-β的中和抗体以及盐酸N（ω）-硝基-L-精氨酸甲酯可显著抑制LPS激活的细胞中HMGB1的释放，但N-[2-（环己氧基）-4-硝基苯基]-甲磺酰胺（NS-398）则不能。HO-1诱导剂、CORM-2或HO-1转染预处理可显著降低LPS激活的细胞中TNF-α、IL-1β和IFN-β的产生。给予HO-1诱导剂或CORM-2可显著降低LPS或CLP攻击的小鼠血浆中HMGB1的水平，同时血清TNF-α和IL-1β水平要么降低（预处理）要么无变化（延迟给药）。无论预处理还是延迟给药，CORM-2和血红素均可使小鼠从LPS或CLP诱导的致死性内毒素血症和脓毒症中获救。综上所述，我们得出结论：HO-1衍生的CO可减少LPS激活的细胞以及LPS或CLP诱导的脓毒症动物模型中HMGB1的释放。
