Connolly E S, Winfree C J, Springer T A, Naka Y, Liao H, Yan S D, Stern D M, Solomon R A, Gutierrez-Ramos J C, Pinsky D J
Department of Neurosurgery, Columbia University, College of Physicians and Surgeons, New York 10032, USA.
J Clin Invest. 1996 Jan 1;97(1):209-16. doi: 10.1172/JCI118392.
Acute neutrophil (PMN) recruitment to postischemic cardiac or pulmonary tissue has deleterious effects in the early reperfusion period, but the mechanisms and effects of neutrophil influx in the pathogenesis of evolving stroke remain controversial. To investigate whether PMNs contribute to adverse neurologic sequelae and mortality after stroke, and to study the potential role of the leukocyte adhesion molecule intercellular adhesion molecule-1 (ICAM-1) in the pathogenesis of stroke, we used a murine model of transient focal cerebral ischemia consisting of intraluminal middle cerebral artery occlusion for 45 min followed by 22 h of reperfusion. PMN accumulation, monitored by deposition of 111In-labeled PMNs in postischemic cerebral tissue, was increased 2.5-fold in the ipsilateral (infarcted) hemisphere compared with the contralateral (noninfarcted) hemisphere (P < 0.01). Mice immunodepleted of neutrophils before surgery demonstrated a 3.0-fold reduction in infarct volumes (P < 0.001), based on triphenyltetrazolium chloride staining of serial cerebral sections, improved ipsilateral cortical cerebral blood flow (measured by laser Doppler), and reduced neurological deficit compared with controls. In wild-type mice subjected to 45 min of ischemia followed by 22 h of reperfusion, ICAM-1 mRNA was increased in the ipsilateral hemisphere, with immunohistochemistry localizing increased ICAM-1 expression on cerebral microvascular endothelium. The role of ICAM-1 expression in stroke was investigated in homozygous null ICAM-1 mice (ICAM-1 -/-) in comparison with wild-type controls (ICAM-1 +/+). ICAM-1 -/- mice demonstrated a 3.7-fold reduction in infarct volume (P < 0.005), a 35% increase in survival (P < 0.05), and reduced neurologic deficit compared with ICAM-1 +/+ controls. Cerebral blood flow to the infarcted hemisphere was 3.1-fold greater in ICAM-1 -/- mice compared with ICAM-1 +/+ controls (P < 0.01), suggesting an important role for ICAM-1 in the genesis of postischemic cerebral no-reflow. Because PMN-depleted and ICAM-1-deficient mice are relatively resistant to cerebral ischemia-reperfusion injury, these studies suggest an important role for ICAM-1-mediated PMN adhesion in the pathophysiology of evolving stroke.
急性中性粒细胞(PMN)募集至缺血后的心脏或肺组织在早期再灌注期具有有害作用,但中性粒细胞流入在进展性中风发病机制中的作用和影响仍存在争议。为了研究PMN是否导致中风后不良神经后遗症和死亡率,并研究白细胞粘附分子细胞间粘附分子-1(ICAM-1)在中风发病机制中的潜在作用,我们使用了一种短暂性局灶性脑缺血小鼠模型,该模型包括大脑中动脉腔内闭塞45分钟,随后再灌注22小时。通过111In标记的PMN在缺血后脑组织中的沉积监测PMN积累,与对侧(未梗死)半球相比,同侧(梗死)半球增加了2.5倍(P < 0.01)。术前免疫清除中性粒细胞的小鼠,基于连续脑切片的氯化三苯基四氮唑染色,梗死体积减少了3.0倍(P < 0.001),同侧皮质脑血流量改善(通过激光多普勒测量),与对照组相比神经功能缺损减轻。在经历45分钟缺血随后22小时再灌注的野生型小鼠中,同侧半球ICAM-1 mRNA增加,免疫组织化学显示脑微血管内皮上ICAM-1表达增加。与野生型对照(ICAM-1 +/+)相比,在纯合缺失ICAM-1小鼠(ICAM-1 -/-)中研究了ICAM-1表达在中风中的作用。与ICAM-1 +/+对照相比,ICAM-1 -/-小鼠梗死体积减少了3.7倍(P < 0.005),存活率提高了35%(P < 0.05),神经功能缺损减轻。与ICAM-1 +/+对照相比,ICAM-1 -/-小鼠梗死半球的脑血流量大3.1倍(P < 0.01),表明ICAM-1在缺血后脑无复流的发生中起重要作用。由于清除PMN和缺乏ICAM-1的小鼠对脑缺血-再灌注损伤相对具有抗性,这些研究表明ICAM-1介导的PMN粘附在进展性中风的病理生理学中起重要作用。