Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia
Reactive oxygen intermediates induce monocyte chemotactic protein-1 in vascular endothelium after brief ischemia
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DOI：
10.1016/s0002-9440(10)62517-5
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发表时间：
2001-10-01
影响因子：
6
通讯作者：
Entman, ML
中科院分区：
文献类型：
--
作者：
Lakshminarayanan, V;Lewallen, M;Entman, ML
趋化因子的表达与心肌梗死再灌注密切相关。在组织坏死、剧烈炎症和炎性细胞因子释放的背景下。再灌注区心肌小静脉特异性合成单核细胞趋化蛋白-1(MCP)-1mRNA的区域与白细胞正常定位的区域相对应。外源可以诱导MCP-1的表达。肿瘤坏死因子(TNF)-α或含有肿瘤坏死因子-α的缺血后心脏淋巴。然而，再灌流早期的肿瘤坏死因子-α的释放并不能解释MCP-1诱导的静脉定位。为了更好地了解介导MCP-1诱导的因素，我们在一个短暂的冠状动脉闭塞模型中研究了缺血/再灌注的作用，在这个模型中，没有看到坏死或炎症反应。成年杂种犬冠状动脉结扎15min，再灌流5h。核糖核酸酶保护实验显示MCP-1mRNA仅在缺血区表达上调。再灌流犬心肌的节段。活性氧清除剂N-(2-巯基丙酰基)-甘氨酸可完全抑制MCP-1的诱导。原位杂交将MCP-1定位于无心肌细胞坏死的小静脉内皮缺血区。GET位移分析显示，缺血区转录因子AP-1和核因子(NF)-kappaB的DNA结合增强，这是MCP-1表达的关键。免疫组织化学染色显示小静脉内皮细胞c-jun和核因子-kappaB(P65)的核移位呈再灌流依赖性，仅在心肌缺血区，N-(2-巯基丙酰基)-甘氨酸可抑制这种移位。体外培养的犬颈静脉内皮细胞用活性氧中间体H_2O_2处理后，单核细胞趋化蛋白-1m RNA水平呈浓度依赖性增加，这种作用可被抗氧化剂N-乙酰-L半胱氨酸抑制，而不能被抗氧化剂N-乙酰-L-半胱氨酸抑制，而吡咯烷二硫代氨基甲酸酯则不能。与我们研究的脑梗塞相比，犬颈静脉内皮细胞的孵育。有缺血后心脏淋巴的细胞不能诱导MCP-1mRNA的表达，这表明在亚致死性缺血期后没有细胞因子介导的MCP-1诱导。这些结果表明，在短暂的缺血发作后，活性氧中间产物的产生能够通过AP-1和NF-kappaB诱导微静脉内皮细胞MCP-1的表达。短时间的缺血/再灌流不足以导致心肌梗死，可诱导MCP-1的表达，潜在地介导缺血非梗死性心脏的血管生成。
Chemokine expression is associated with reperfusion of infarcted myocardium. in the setting of tissue necrosis, intense inflammation, and inflammatory cytokine release. The specific synthesis of monocyte chemotactic protein (MCP)-1 mRNA by cardiac venules in reperfused infarcts corresponded to the region where leukocytes; normally localize. MCP-1 could be induced by exogenous. tumor necrosis factor (TNF)-alpha or by postischemic cardiac lymph containing TNF-alpha. However, the release of TNF-alpha during early reperfusion did not explain the venular localization of MCP-1 induction. To better understand the factors mediating MCP-1 induction, we examined the role of ischemia/reperfusion in a model of brief coronary occlusion in which no necrosis or inflammatory response is seen. Adult mongrel dogs were subjected to 15 minutes of coronary occlusion and 5 hours of reperfusion. Ribonuclease protection assay revealed up-regulation of MCP-1 mRNA only in ischemic. segments of reperfused canine myocardium. Pretreatment with the reactive oxygen scavenger N-(2-mercaptopropionyl)-glycine completely inhibited MCP-1 induction. In situ hybridization localized MCP-1 message to small venular endothelium ischemic areas without myocyte necrosis. Get shift analysis of nuclear extracts from the ischemic area showed enhanced DNA binding of the transcription factors AP-1 and nuclear factor (NF)-kappaB, crucial for MCP-1 expression, in ischemic myocardial regions. Immunohistochemical staining demonstrated reperfusion-dependent nuclear translocation of c-Jun and NF-kappaB (p65) in small venular endothelium., only in the ischemic regions of the myocardium, that was inhibited by N-(2-mercaptopropionyl)-glycine. in vitro, treatment of cultured canine jugular vein endothelial cells with the reactive oxygen intermediate H2O2 induced a concentration-dependent increase in MCP-1 mRNA levels, which was inhibited by the antioxidant N-acetyl-L-cysteine, a precursor of glutathione, but not pyrrolidine dithiocarbamate, an inhibitor of NF-kappaB and activator of AP-1. In contrast to our studies with infarction, incubation of canine jugular vein endothelial. cells with postischemic cardiac lymph did not induce MCP-1 mRNA expression suggesting the absence of cytokine-mediated MCP-1 induction after a sublethal ischemic period. These results suggest that reactive oxygen intermediate generation, after a brief ischemic episode, is capable of inducing MCP-1 expression in venular endothelium through AP-1 and NF-kappaB. Short periods of ischemia/reperfusion, insufficient to produce a myocardial infarction, induce MCP-1 expression, potentially mediating angiogenesis in the ischemic noninfarcted heart.