大鼠急性脑缺血模型中基质金属蛋白酶9和转化生长因子β_1的表达及其组织来源分析  

Expression and histogenesis of matrix metalloproteinase-9 and transforming growth factor-beta 1 in acute cerebral ischemia model of rats

作　　者：王艳国[1] 李得春[2] 胡海[1] 张圆[1] 

机构地区：[1]包头医学院病理生理学教研室,内蒙古包头014060 [2]包头医学院第一附属医院检验科,内蒙古包头014060

出　　处：《中国病理生理杂志》2014年第10期1789-1793,共5页Chinese Journal of Pathophysiology

基　　金：内蒙古自治区教育厅高等学校科学研究项目(No.NJZY12224)

摘　　要：目的：探讨大鼠急性缺血模型中基质金属蛋白酶9（matrix metalloproteinase 9,MMP-9）和转化生长因子β1（transforming growth factorβ1,TGF-β1）的表达及其组织来源。方法：280～330 g雄性Wistar大鼠,使用线栓法建立急性大鼠脑缺血模型,并采用Zealonga 5分制评分法评定模型建立的效果。分别取正常对照组、假处理组以及缺血后6 h、12 h、1 d、2 d、6 d和14 d组大鼠的大脑皮质和海马,采用定量PCR检测脑组织中MMP-9和TGF-β1的表达。取对应时点各组大鼠大脑进行MMP-9和TGF-β1的原位组织化学染色。取对应时点各组大鼠血浆,以酶联免疫法（ELISA）检测MMP-9和TGF-β1的浓度变化。结果：定量PCR结果显示MMP-9 mRNA水平在急性缺血6h即开始升高,到急性缺血2 d时最高。而TGF-β1mRNA水平在急性缺血12 h开始升高,到急性缺血6 d时达到最高水平。与假处理组样本相比,缺血时间为12 h、1 d、2 d、6 d和14 d的样本有显著差异（P〈0.05）。原位组织化学染色结果表明从急性脑缺血后1 d起,大脑的皮质和海马区域开始有MMP-9表达。急性脑缺血后2 d左右,大脑皮质的MMP-9染色最明显。在急性缺血2 d后皮质和海马区域开始能观察到TGF-β1表达,6 d时最明显。ELISA结果显示缺血后12 h,血浆中即可检测到MMP-9和TGF-β1的表达。与假处理组相比,缺血1 d时2种因子的表达差异显著（P〈0.05）。结论：大鼠急性脑缺血模型中其脑组织是急性脑缺血后MMP-9和TGF-β1的来源之一,该发现为进一步开展相关研究提供了线索。AIM： To observe the expression and tissue localization of matrix metalloproteinase9（ MMP-9） and transforming growth factor beta 1（ TGF-β1） in the rat acute cerebral ischemia model. METHODS： Male Wistar rats were used to establish acute cerebral ischemia model by a suturing method. The rats were divided into normal control group,sham group and ischemia 6 h,12 h,1 d,2 d,6 d and 14 d groups. The rat cerebral cortex and hippocampus of the brain were collected at different time points. The mRNA and protein levels of MMP-9 and TGF-β1in the brain tissues were detected by real-time PCR and in situ histochemistry staining,respectively. The levels of MMP-9 and TGF-β1in the plasma were also measured by ELISA. RESULTS： The results of real-time PCR showed that the mRNA levels of MMP-9 began to increase 6 h after acute ischemia and reached to a peak 2 d after acute ischemia. Similarly,the mRNA level of TGF-β1began to rise 12 h after acute ischemia and reached to the highest level 6 d after acute ischemia. Compared with the sham rats,the mRNA levels of MMP-9 and TGF-β1in the rat brains that collected at ischemic time of 12 h,1 d,2 d,6 d and 14 d were significantly increased. Moreover,results of in situ histochemical staining showed that the expression of MMP-9 was detected at cerebral cortex and hippocampus 1 d after acute cerebral ischemia. Further studies showed that MMP-9 dyeing of the rat cerebral cortex was most obvious 2 d after the acute cerebral ischemia. Similarly,the rat cortex and hippocampus began to express TGF-β12 d after acute ischemia and TGF-β1staining at rat cerebral cortex was most obvious 6 d after the acute cerebral ischemia. In addition,ELISA showed that the increase in MMP-9 and TGF-β1was detected in the plasma 12 h after ischemia. Compared with the sham rats,the level of these 2 factors significantly upregulated since 1 d after ischemia.CONCLUSION： The brain tissue itself contributes to the upregulation of MMP-9 and TGF-β1post acute cerebral ischemia,which shed light on the relat

关 键 词：基质金属蛋白酶9 转化生长因子Β1 急性脑缺血 

分 类 号：R363[医药卫生—病理学]