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缺血再灌注损伤大鼠脑组织MMP-2、MMP-9动态变化与脑水肿改变
焦海霞1,2,王萍1,马腾1,薛一雪1*
0
(1.中国医科大学基础医学院神经生物学教研室, 沈阳 110001; 2.福建医科大学基础医学院生理学与病理生理学系生理学教研室,福州 350004)
摘要:
[摘要]目的探讨发生脑缺血再灌注损伤时MMP-2、MMP-9表达和活性的变化规律及应用强力霉素后脑水肿的变化。方法大脑中动脉线栓法制备局灶性脑缺血再灌注模型,根据再灌注时间不同分为再灌注3、6、12、24、72、120 h组及假手术组。应用干湿重法测定强力霉素干预后缺血侧脑半球含水量的变化,应用蛋白质印迹法检测缺血侧脑组织MMP-2和MMP-9蛋白表达变化,应用明胶酶谱法检测缺血侧脑组织MMP-2和MMP-9活性变化。结果MMP-2在脑缺血再灌注3 h和120 h表达和活性升高(P<0.01);MMP-9在脑缺血再灌注6 h表达和活性开始升高(P<0.01),到再灌注24 h达到峰值,再灌注120 h降至正常水平(P>0.05);脑缺血再灌注大鼠各时间点缺血侧脑半球含水量与假手术组相比皆升高,并且随着再灌注时间延长持续升高;应用强力霉素大鼠缺血侧脑半球含水量与同时间点生理盐水组相比降低(P<0.05或P<0.01)。结论MMP-2和MMP-9降解细胞外基质引发血管源性脑水肿,MMP-2和MMP-9的表达及其活性在脑缺血再灌注120 h内的交替变化可能会影响脑水肿的发展。
关键词:  脑缺血  再灌注损伤  基质金属蛋白酶  脑水肿  强力霉素
DOI:10.3724/SP.J.1008.2010.01193
投稿时间:2010-07-15修订日期:2010-10-08
基金项目:国家自然科学基金(30872656,30973079),教育部高等学校博士学科点专项科研基金(20092104110015),沈阳市科学技术计划项目(1091175-1-01, 1081266-9-00),福建省自然科学基金(C0610022).
Changes of MMP-2, MMP-9 contents in cerebral tissues and cerebral edema in cerebral ischemia-reperfusion rats
JIAO Hai-xia1,2, WANG Ping1, MA Teng1, XUE Yi-xue1 *
(1. Department of Neurobiology, College of Basic Medicine, China Medical University, Shenyang 110001, Liaoning, China; 2. Department of Physiology and Pathophysiology, College of Basic Medicine, Fujian Medical University, Fuzhou 350004, Fujian, China)
Abstract:
[Abstract]ObjectiveTo investigate the expression and activation of MMP-2, MMP-9 in cerebral tissues of cerebral ischemia-reperfusion rats and the effects of doxycycline on the cerebral edema. MethodsCerebral ischemia-reperfusion model was established by middle cerebral artery occlusion (MCAO). The rats were randomly divided into a sham-operated and 6 reperfusion groups according to different reperfusion periods (3 h, 6 h, 12 h, 24 h, 72 h, and 120 h groups). Dry weight method was used to evaluate hemispheric water content of the ischemic side after treatment with doxycycline. Western blotting analysis and Gelatin zymography were used to determine the expression and activity of MMP-2 and MMP-9 in the ischemic cerebral tissues. MethodsThe protein expression and activity of MMP-2 were significantly increased in the ischemia tissue at 3 h and 120 h after reperfusion(P<0.01); the protein expression and activity of MMP-9 began to increase at 6 h after reperfusion(P<0.01), reached the peak at 24 h, and then returned to the basal levels at 120 h after reperfusion (P>0.05). The hemispheric water content of the ischemic side gradually increased with reperfusion periods compared with the sham-operated group. Dexycycline significantly reduced the hemispheric water contents compared with the normal saline at the same time points(P<0.05 or P<0.01). MethodsMMP-2, MMP-9 can induce vasogenic brain edema by degrading basal membrane of capillary vessel. The alteration of MMP-2, MMP-9 expression and activities are likely associated with brain edema during cerebral ischemia-reperfusion.
Key words:  brain ischemia  reperfusion injury  matrix metalloproteinases  brain edema  doxycycline