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。 2009年11月;15(11):1318-21.
DOI：10.1038 / nm.2053。 EPUB 2009年10月25日。

细胞外组蛋白是败血症中死亡的主要介质

徐徐1, 小美张, 罗萨纳佩莱奥, Marc勋爵, Concetta T Ammollo, Fabrizio Semeraro, 弗莱彻B泰勒, naomi l esm, 佛罗里亚卢普, 查尔斯T ESMON.
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细胞外组蛋白是败血症中死亡的主要介质

徐徐et al. Nat Med。 2009年11月。
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抽象

Hyperinflammatory responses can lead to a variety of diseases, including sepsis. We now report that extracellular histones released in response to inflammatory challenge contribute to endothelial dysfunction, organ failure and death during sepsis. They can be targeted pharmacologically by antibody to histone or by activated protein C (APC). Antibody to histone reduced the mortality of mice in lipopolysaccharide (LPS), tumor necrosis factor (TNF) or cecal ligation and puncture models of sepsis. Extracellular histones are cytotoxic toward endothelium in vitro and are lethal in mice. In vivo, histone administration resulted in neutrophil margination, vacuolated endothelium, intra-alveolar hemorrhage and macro- and microvascular thrombosis. We detected histone in the circulation of baboons challenged with Escherichia coli, and the increase in histone levels was accompanied by the onset of renal dysfunction. APC cleaves histones and reduces their cytotoxicity. Co-infusion of APC with E. coli in baboons or histones in mice prevented lethality. Blockade of protein C activation exacerbated sublethal LPS challenge into lethality, which was reversed by treatment with antibody to histone. We conclude that extracellular histones are potential molecular targets for therapeutics for sepsis and other inflammatory diseases.

数据

Fig.1
Fig.1
Cytotoxicity of extracellular histones toward endothelium and APC cleavage of histones. (a) EA.hy926 cells were cultured with calf thymus histones (50µg ml−1) or calf thymus histone H1, H2A, H2B, H3 or H4 (20µg ml−1）37°C的1小时。通过流式细胞术进行PI染色测量细胞损伤。（b）在与上述测定中的组蛋白，H3或H4孵育期间不存在或存在APC（100nm）。（c）纯化小牛胸腺H3（顶面板）或H4（底板）的SDS-PAGE分析（100μgmL−1）在37℃下用指定的人APC孵育1小时。（d）SDS-PAGE分析纯化的小牛胸腺组蛋白H3（顶面板）或H4（底板）（100μgmL−1）在没有或存在0.5mg ml的情况下孵育10nm人APC−1PS / PC或PE / PS / PC脂质体在37°C时为1小时。
Fig.2
Fig.2
APC切割组蛋白in vitro和体内。（a）在37℃下在37℃下在不存在或存在下的表明浓度下，用CALF胸腺组蛋白培养EA.hy926细胞。通过流式细胞术测量细胞损伤，用于PI染色并表示为平均荧光指数（MFI）。（b）小牛胸腺组蛋白（50μgml−1）在37℃下与APC（100nm）孵育的时间，然后与PPACK（10μm）混合以灭活APC。上述培养基用于培养EA.HY926细胞1小时进行细胞毒性测定或（c）对SDS-PAGE进行的（c）进行SDS-PAGE和Western印迹，在时刻表示的H3或H4。（d）用小牛胸腺组蛋白培养EA.hy926细胞（50μgml−1）在37℃下在不存在或存在蛋白C（100nm），凝血酶（T）（10nm）或Apc（100nm），持续30分钟。通过流式细胞术进行PI染色测量细胞损伤。（e）狒狒等离子体样品H3的Western印迹分析在后面的时间大肠杆菌要么大肠杆菌加上APC挑战。（f）在人体化学患者的APC治疗开始时显示的血浆样品H3的Western印迹分析。
Fig.3
Fig.3
静脉内注射组蛋素引发炎症和细胞损伤反应。(a) Survival rates of mice injected intravenously with calf thymus histones (75 mg per kg) with or without APC (5 mg per kg). (b–d) Pathological changes of mouse lung three hours after intravenous injection of histones (50 mg per kg). Immunofluorescence staining for neutrophil elastase detected massive neutrophil accumulation in the alveolar microvasculature (c: histone treated vs. b: control). Alveolar capillaries are almost fully obstructed by cells, as seen by electron microscopy (d: PMN). (e–f) Histones induce strong alterations of the selective permeability of plasma membranes and subsequent intracellular edema, vacuolization (*) of the intracellular organelles (endoplasmic reticulum, Golgi, and mitochondria), both within endothelial (EC) and type I epithelial cells (ep–I). av, alveolae; cav, caveolae; RBC, red blood cells. Magnification bars: b and c: 50 µm; d, 10 µm; e and f, 500 nm.
Fig.4
Fig.4
Antibody to H4 protects mice from the lethality of LPS, CLP and TNF体内。（a）小鼠用高剂量的LPS（每千克/克）静脉内注射小鼠，与H4或小鼠IgG对照抗体（每kg 20mg）。指出了每组的存活率。（b）小鼠用低剂量的LPS（每千克/克）静脉内注射或不含蛋白C（每千克/克）的抗体，并抗体至H4或H2B（每kg 20mg）。指出了每组的存活率。在高剂量的LPS（每kg）加上H4或小鼠IgG对照抗体（每千克/克）或（D）后6小时后，小鼠等离子体的H3（C）24小时的蛋白质印迹分析具有或不具有蛋白C的抗体（每千克/ kg的2.5mg）的LPS（每千克/克/毫升）。（e）对CLP进行的小鼠的存活率，然后用抗体处理到H4或小鼠IgG对照抗体（20mg / kg）加庆大霉素（每公斤5mg）6小时后CLP。（f）静脉注射的小鼠的存活率与TNF（0.75mg / kg）加上H4或小鼠IgG对照抗体（每kg 5mg）。
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评论

  • 败血症：组蛋白的黑暗面。
    Chaput C，Zychlinsky A. Chaput C等人。 Nat Med。2009年11月15日（11）：1245-6。DOI：10.1038 / NM1109-1245。 Nat Med。200.9. PMID：19893552 没有抽象可用。

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