In situ determination by surface chemiluminescence of temporal relationships between evolving warm ischemia-reperfusion injury in rat liver and phagocyte activation and recruitment
In situ determination by surface chemiluminescence of temporal relationships between evolving warm ischemia-reperfusion injury in rat liver and phagocyte activation and recruitment
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DOI：
10.1002/hep.510310312
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发表时间：
2000-03-01
期刊：
影响因子：
13.5
通讯作者：
Poli, G
中科院分区：
文献类型：
--
作者：
Cutrìn, JC;Boveris, A;Poli, G
肝脏缺血再灌注的特征是氧依赖性自由基链反应速率增加和活性氧稳态浓度增加。本研究的目的是评估在原位产生的活性氧自由基及其与吞噬细胞的激活和再灌注大鼠肝脏募集的关系。大鼠肝脏进行2小时的选择性小叶缺血和再灌注长达12小时。测定了以下参数：原位肝脏化学发光，理解为反映单线态氧（O-1（2））的组织稳态浓度;髓过氧化物酶组织活性;中性粒细胞数量;和坏死程度。在血液回流30分钟后测量早期化学发光爆发（氧化应激的早期阶段），随后在再灌注4至12小时后复发并进一步增加（氧化应激的晚期阶段）。早期和晚期相均通过氯化钆（GdCl 3）预处理进行了修改，指出了枯否细胞的关键作用。中性粒细胞浸润到肝脏，髓过氧化物酶活性，原位化学发光和坏死被认为是强相关的4至12小时的再灌注期间（r = 0.960;平均的4个相关系数）。与常驻吞噬细胞一起，中性粒细胞募集和活化似乎对氧依赖性自由基反应的增加和肝脏再灌注损伤的放大做出了重大贡献。表面化学发光似乎适当地描述了在原位和在体内进行性组织的急性炎症反应与吞噬细胞介导的肝损伤。
Liver ischemia-reperfusion is characterized by an increased oxygen-dependent free radical chain-reaction rate and an increased steady-state concentration of reactive oxygen species. The aim of this study was to evaluate the in situ generation of reactive oxygen species and its relationship with phagocyte activation and recruitment in reperfused rat liver. Rat livers were subjected to 2 hours of selective lobular ischemia and reperfusion for up to 12 hours. The following parameters were determined: in situ liver chemiluminescence, understood to reflect the tissue steady-state concentration of singlet oxygen (O-1(2)); myeloperoxidase tissue activity; the number of neutrophils; and the degree of necrosis. An early chemiluminescence burst was measured after 30 minutes of blood reflow (early phase of oxidative stress), followed by a relapse and a further increase after 4 to 12 hours of reperfusion (late phase of oxidative stress). Both early and late phases were modified by pretreatment with gadolinium chloride (GdCl3), pointing to a key role of the Kupffer cells. Neutrophils infiltrated into the liver, myeloperoxidase activity, in situ chemiluminescence, and necrosis were found to be strongly correlated over the 4- to 12-hour reperfusion period (r = .960; average of the 4 correlation coefficients). Together with resident phagocytes, neutrophil recruitment and activation appear to provide a major contribution to the increase of oxygen-dependent free-radical reactions and amplification of liver reperfusion damage. Surface chemiluminescence appears to properly describe the in situ and in vivo progressive organization of the acute inflammatory response with phagocyte-mediated liver injury.