Kelly Melissa A, Shuaib Ashfaq, Todd Kathryn G
Center for Neuroscience, University of Alberta, Edmonton, AB, Canada.
Exp Neurol. 2006 Jul;200(1):38-49. doi: 10.1016/j.expneurol.2006.01.032. Epub 2006 Apr 19.
Thrombolysis with tissue plasminogen activator (tPA) is the only pharmacotherapy available for cerebral ischemia. However, the use of tPA can increase the risk of hemorrhage due to blood-brain barrier (BBB) breakdown. Recent evidence suggests that increased activation of matrix metalloproteinases (MMPs) may be involved in this breakdown. This study examines the temporal profile of MMP-2 and -9 following tPA administration to ischemic rats. Male Sprague-Dawley rats were randomly assigned to one of four groups (Sham-tPA; Sham-Saline; Ischemia-tPA; Ischemia-Saline; group n = 6, total N = 120). Focal embolic ischemia was induced by middle cerebral artery occlusion through injection of an autologous clot. One hour post-surgery, tPA (10 mg/kg) or saline was delivered intravenously and animals were euthanized at 3, 6, 12, or 24 h after onset of ischemia. Infarct volume was measured by TTC staining; BBB components examined immunohistochemically; and MMP activation measured by gelatin zymography. Our results show that tPA significantly reduced infarct volumes (overall infarct volume-Sham-tPA: 5.80 +/- 4.55 [mean +/- SE]; Sham-Saline: 5.00 +/- 4.23; Ischemia-tPA: 186.1 +/- 73.45; Ischemia-Saline: 284.8 +/- 88.74; all P < 0.05). Treatment with tPA was also associated with the activation of MMP-9 at 6, 12, and 24 h following ischemia. No temporal changes were observed in MMP-2 activation, although tPA administration increased its activity compared to saline treatment. Analyses of immunohistochemistry showed that destruction of components of the BBB followed MMP-9 activation. Thus, increased MMP-9 activation may, in part, be responsible for the increases in hemorrhagic transformation reported with use of tPA. Our study is the first to demonstrate the temporal profile of MMP activation following thrombolysis with tPA in a model of thrombotic focal cerebral ischemia.
使用组织型纤溶酶原激活剂(tPA)进行溶栓是目前治疗脑缺血唯一可用的药物疗法。然而,使用tPA会因血脑屏障(BBB)破坏而增加出血风险。最近的证据表明,基质金属蛋白酶(MMPs)激活增加可能与这种破坏有关。本研究考察了给缺血大鼠注射tPA后MMP - 2和 - 9随时间的变化情况。将雄性Sprague - Dawley大鼠随机分为四组之一(假手术 - tPA组;假手术 - 生理盐水组;缺血 - tPA组;缺血 - 生理盐水组;每组n = 6,共N = 120)。通过注射自体血凝块诱导大脑中动脉闭塞,造成局灶性栓塞性缺血。术后1小时,静脉注射tPA(10 mg/kg)或生理盐水,缺血发作后3、6、12或24小时对动物实施安乐死。通过TTC染色测量梗死体积;采用免疫组织化学方法检测血脑屏障成分;通过明胶酶谱法测量MMP激活情况。我们的结果显示,tPA显著减小了梗死体积(总体梗死体积 - 假手术 - tPA组:5.80±4.55 [平均值±标准误];假手术 - 生理盐水组:5.00±4.23;缺血 - tPA组:186.1±73.45;缺血 - 生理盐水组:284.8±88.74;所有P < 0.05)。tPA治疗还与缺血后6、12和24小时MMP - 9的激活有关。尽管与生理盐水治疗相比,tPA给药增加了MMP - 2的活性,但未观察到MMP - 2激活随时间的变化。免疫组织化学分析表明,血脑屏障成分的破坏发生在MMP - 9激活之后。因此,MMP - 9激活增加可能部分导致了使用tPA后报道的出血性转化增加。我们的研究首次在血栓性局灶性脑缺血模型中展示了tPA溶栓后MMP激活随时间的变化情况。