中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
10期
1789-1793
,共5页
王艳国%李得春%胡海%张圆
王豔國%李得春%鬍海%張圓
왕염국%리득춘%호해%장원
基质金属蛋白酶9%转化生长因子β1%急性脑缺血
基質金屬蛋白酶9%轉化生長因子β1%急性腦缺血
기질금속단백매9%전화생장인자β1%급성뇌결혈
Matrix metalloproteinase 9%Transforming growth factor beta 1%Acute brain ischemia
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6 h即开始升高,到急性缺血2 d时最高。而TGF-β1 mRNA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。
目的:探討大鼠急性缺血模型中基質金屬蛋白酶9(matrix metalloproteinase 9,MMP-9)和轉化生長因子β1( transforming growth factorβ1,TGF-β1)的錶達及其組織來源。方法:280~330 g雄性Wistar大鼠,使用線栓法建立急性大鼠腦缺血模型,併採用Zealonga 5分製評分法評定模型建立的效果。分彆取正常對照組、假處理組以及缺血後6 h、12 h、1 d、2 d、6 d和14 d組大鼠的大腦皮質和海馬,採用定量 PCR檢測腦組織中MMP-9和TGF-β1的錶達。取對應時點各組大鼠大腦進行MMP-9和TGF-β1的原位組織化學染色。取對應時點各組大鼠血漿,以酶聯免疫法( ELISA)檢測MMP-9和TGF-β1的濃度變化。結果:定量PCR結果顯示MMP-9 mRNA水平在急性缺血6 h即開始升高,到急性缺血2 d時最高。而TGF-β1 mRNA水平在急性缺血12 h開始升高,到急性缺血6 d時達到最高水平。與假處理組樣本相比,缺血時間為12 h、1 d、2 d、6 d和14 d的樣本有顯著差異(P<0.05)。原位組織化學染色結果錶明從急性腦缺血後1 d起,大腦的皮質和海馬區域開始有MMP-9錶達。急性腦缺血後2 d左右,大腦皮質的MMP-9染色最明顯。在急性缺血2 d後皮質和海馬區域開始能觀察到TGF-β1錶達,6 d時最明顯。 ELISA結果顯示缺血後12 h,血漿中即可檢測到MMP-9和TGF-β1的錶達。與假處理組相比,缺血1 d時2種因子的錶達差異顯著(P<0.05)。結論:大鼠急性腦缺血模型中其腦組織是急性腦缺血後MMP-9和TGF-β1的來源之一,該髮現為進一步開展相關研究提供瞭線索。
목적:탐토대서급성결혈모형중기질금속단백매9(matrix metalloproteinase 9,MMP-9)화전화생장인자β1( transforming growth factorβ1,TGF-β1)적표체급기조직래원。방법:280~330 g웅성Wistar대서,사용선전법건립급성대서뇌결혈모형,병채용Zealonga 5분제평분법평정모형건립적효과。분별취정상대조조、가처리조이급결혈후6 h、12 h、1 d、2 d、6 d화14 d조대서적대뇌피질화해마,채용정량 PCR검측뇌조직중MMP-9화TGF-β1적표체。취대응시점각조대서대뇌진행MMP-9화TGF-β1적원위조직화학염색。취대응시점각조대서혈장,이매련면역법( ELISA)검측MMP-9화TGF-β1적농도변화。결과:정량PCR결과현시MMP-9 mRNA수평재급성결혈6 h즉개시승고,도급성결혈2 d시최고。이TGF-β1 mRNA수평재급성결혈12 h개시승고,도급성결혈6 d시체도최고수평。여가처리조양본상비,결혈시간위12 h、1 d、2 d、6 d화14 d적양본유현저차이(P<0.05)。원위조직화학염색결과표명종급성뇌결혈후1 d기,대뇌적피질화해마구역개시유MMP-9표체。급성뇌결혈후2 d좌우,대뇌피질적MMP-9염색최명현。재급성결혈2 d후피질화해마구역개시능관찰도TGF-β1표체,6 d시최명현。 ELISA결과현시결혈후12 h,혈장중즉가검측도MMP-9화TGF-β1적표체。여가처리조상비,결혈1 d시2충인자적표체차이현저(P<0.05)。결론:대서급성뇌결혈모형중기뇌조직시급성뇌결혈후MMP-9화TGF-β1적래원지일,해발현위진일보개전상관연구제공료선색。
AIM:To observe the expression and tissue localization of matrix metalloproteinase 9 (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-β1 in the brain tissues were detec-ted by real-time PCR and in situ histochemistry staining, respectively.The levels of MMP-9 and TGF-β1 in the plasma were also measured by ELISA.RESULTS:The results of real-time PCR showed that the mRNA levels of MMP-9 began to in-crease 6 h after acute ischemia and reached to a peak 2 d after acute ischemia.Similarly, the mRNA level of TGF-β1 began 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-β1 in 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-β1 2 d after acute ischemia and TGF-β1 staining 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-β1 was 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-β1 post acute cerebral ischemia, which shed light on the related research in the field.