AN INVITRO MODEL OF ISCHEMIA REPERFUSION-INDUCED MICROVASCULAR INJURY
AN INVITRO MODEL OF ISCHEMIA REPERFUSION-INDUCED MICROVASCULAR INJURY
复制标题
DOI：
10.1152/ajpgi.1990.259.1.g134
复制
发表时间：
1990-07-01
影响因子：
--
通讯作者：
KVIETYS, PR
中科院分区：
文献类型：
--
作者：
INAUEN, W;GRANGER, DN;KVIETYS, PR
本研究的主要目的是建立缺血/再灌注（I/R）诱导的微血管损伤的体外模型。将培养的静脉内皮细胞培养至融合后，用51Cr标记，并暴露于不同的缺氧时间（0.5、1、2、3和4 h）。在再氧后的不同时间（1、2、4、6、8和18 h），检测51Cr释放和细胞脱离（细胞损伤指标）。由于体内研究表明中性粒细胞与I/R损伤有关，在一些实验中，人中性粒细胞在再氧化后加入内皮细胞。缺氧期。2 h后释放70-80% 51Cr， 80-95%细胞脱离。在这些条件下（接近最大损伤），添加中性粒细胞产生的影响可以忽略不计。短时间的缺氧。1 h 51Cr释放30-40%，细胞脱离50-60%。在这些条件下（中度细胞损伤），添加中性粒细胞增强内皮细胞损伤。通过30分钟的缺氧时间，我们还评估了超氧化物歧化酶（SOD; 300 U/ml）和别嘌呤醇(20 .mu。M)在中性粒细胞存在或不存在的情况下对缺氧/再氧化（A/R）诱导的损伤的影响。在没有中性粒细胞的情况下，SOD或丙烯醇对A/ r诱导的损伤没有保护作用。然而，在中性粒细胞的存在下，SOD和丙烯醇都减弱了51Cr释放的增加。使用这种体外I/R损伤模型得出的结果与已发表的体内研究基本一致。因此，这个体外模型可以为I/R损伤的机制提供进一步的见解。
The major objective of this study was to develop an in vitro model of ischemia/reperfusion (I/R)-induced microvascular injury. Cultured venular endothelial cells were grown to confluency, labeled with 51Cr, and exposed to different durations of anoxia (0.5, 1, 2, 3, and 4 h). 51Cr release and cell detachment (indexes of cell injury) were determined at different times after reoxygenation (1, 2, 4, 6, 8, and 18 h). Because in vivo studies have implicated neutrophils in I/R injury, in some experiments human neutrophils were added to the endothelial cells upon reoxygenation. Periods of anoxia .gtoreq. 2 h resulted in 70-80% 51Cr release and 80-95% cell detachment upon reoxygenation. Under these conditions (near maximal injury), the addition of neutrophils produced negligible effects. Periods of anoxia .ltoreq. 1 h resulted in 30-40% 51Cr release and 50-60% cell detachment. Under these conditions (moderate cell injury), addition of neutrophils enhanced endothelial cell injury. Using a 30-min period of anoxia, we also assessed the effects of superoxide dismutase (SOD; 300 U/ml) and allopurinol (20 .mu.M) on anoxia/reoxygenation (A/R)-induced injury in the presence or absence of neutrophils. In the absence of neutrophils, SOD or alluprinol did not protect against A/R-induced injury. However, in the presence of neutrophils, both SOD and alluprinol attenuated the increases in 51Cr release. The results derived using this in vitro model of I/R injury are largely consistent with published in vivo studies. Thus this in vitro model may provide further insights regarding the mechanisms involved in I/R injury.