Schumer M, Colombel M C, Sawczuk I S, Gobé G, Connor J, O'Toole K M, Olsson C A, Wise G J, Buttyan R
Division of Urology, Maimonides Medical Center, Brooklyn.
Am J Pathol. 1992 Apr;140(4):831-8.
A multiparametric analysis to demonstrate that even brief periods of arterial clamping can initiate extensive cell loss in a rat kidney through the process of apoptosis during the 48-hour period after reperfusion was performed. Microscopic examination of rat renal tissues subject to a 5-, 30-, or 45-minute period of complete ischemia showed the presence of apoptotic bodies both within and occasionally between renal tubules, appearing as early 12 hours after reperfusion, and increasing in numbers at 24 hours. Furthermore, DNA extracted from such reperfused renal tissue demonstrated the appearance of a distinct "ladder" pattern of DNA fragments after electrophoresis in agarose gels, a phenomenon commonly associated with cells undergoing apoptosis and in contrast to the predominant smear pattern obtained after electrophoresis of DNA extracted from necrotic renal tissue. Finally, messenger RNA (mRNA) encoding sulfated glycoprotein-2, a gene product previously identified to apoptotic renal cells, was found to be highly expressed in the 30-minute arterial clamped rat kidney after 24 hours of reperfusion, but was not detectable in mRNA extracted from renal tissue after 24 hours chronic infarction. This study demonstrates that a combination of morphologic, biochemical, and molecular markers can be used to distinguish predominant modes of cell death in varying forms of tissue injury. Application of these analytical techniques to renal vascular injury has distinguished that brief periods of complete ischemia initiates a form of cell death (apoptosis) during a subsequent reperfusion phase that is drastically different from cellular necrosis induced by prolonged severe ischemia.
进行了一项多参数分析，以证明即使是短暂的动脉钳夹也会在再灌注后的48小时内通过凋亡过程引发大鼠肾脏广泛的细胞损失。对经历5分钟、30分钟或45分钟完全缺血的大鼠肾组织进行显微镜检查发现，肾小管内及偶尔在肾小管之间存在凋亡小体，在再灌注后12小时早期出现，并在24小时时数量增加。此外，从这种再灌注肾组织中提取的DNA在琼脂糖凝胶电泳后显示出明显的DNA片段“梯状”模式，这一现象通常与经历凋亡的细胞相关，与从坏死肾组织中提取的DNA电泳后得到的主要涂片模式形成对比。最后，编码硫酸化糖蛋白-2的信使核糖核酸（mRNA），一种先前已确定与凋亡肾细胞相关的基因产物，在再灌注24小时后的30分钟动脉钳夹大鼠肾脏中高度表达，但在慢性梗死24小时后的肾组织提取的mRNA中未检测到。这项研究表明，形态学、生化和分子标记物的组合可用于区分不同形式组织损伤中细胞死亡的主要模式。将这些分析技术应用于肾血管损伤已表明，短暂的完全缺血会在随后的再灌注阶段引发一种细胞死亡形式（凋亡），这与长时间严重缺血诱导的细胞坏死截然不同。
