脑缺血再灌注后热休克蛋白70、c-fos的表达及柘树制剂的神经保护作用  

Expression of HSP70,c-fos following focal cerebral ischemic reperfusion in rats and neuroprotective effect of Cudrania tricuspidata

作　　者：孙琳[1] 刘爱芬[2] 李义召[2] 陈立云[2] 王晓云[1] 韩丹春[2] 

机构地区：[1]山东大学第二医院神经内科,济南250033 [2]齐鲁医院神经内科

出　　处：《临床神经病学杂志》2005年第5期354-356,共3页Journal of Clinical Neurology

基　　金：山东省科委资助项目(课题编号973000056)

摘　　要：目的观察局灶脑缺血再灌注后热休克蛋白(HSP)70、c-fos的表达及其与细胞调亡的关系,探讨柘树制剂对脑缺血后神经细胞损伤的保护作用。方法采用改良Longa法制作大鼠局灶脑缺血再灌注模型。柘树制剂预处理组(柘树组)大鼠在实验前灌服柘树制剂2 m l每日3次,连用5 d。在缺血再灌注不同时点(1 h、6 h、12 h、24 h、3 d、7 d)将大鼠处死取脑,进行HSP70及c-fos免疫组化染色、c-fos mRNA原位杂交、原位末端标记(TUNEL)及HE染色,并对其阳性结果进行半定量分析。结果脑缺血再灌注能诱导HSP70及c-fos的表达。缺血再灌注6 h组HSP70在缺血侧皮质及基底节开始表达,24 h达高峰。缺血再灌注1 h组c-fos即有表达,6 h达高峰,后逐渐下降。细胞凋亡于缺血再灌注6 h最显著。柘树组HSP70及c-fos表达的阳性细胞数均较缺血再灌注组明显增加,两组比较差异均有显著性(均P<0.01),而TUNEL阳性细胞数明显减少。结论HSP70及c-fos均参与了脑缺血的病理生理过程,柘树制剂对脑缺血再灌注损伤有保护作用。Objective To investigate the expression of heat shock protein 70 （HSP70） and c-los, the presence of apoptosis and the ueuroprolective effects of Cudrania tricuspidata root extrate （Ecr） after focal cerebral ischemic reperfusion in rats. Methods Models of middle cerebral artery, occlusion （MCAO） were used in this study. The rats in Ecr pretreated ischemia reperfusion （IR） group were fed with Ecr （2 ml tid） for 5 days before MCAO. The brain specimen were took at different reperfusion limes： 1 h, 6 h, 12 h, 24 h, 3 d and 7 d after recirculation. Then immunohistnchemistry, insitu hybridization, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） were used to detect the expression of HSPTO, c-fos gene and cell apoptosis in the brains. Positive results were semiquantitatively analyzed. Results IR could induce the expression of HSP70 and e-fos. HSP70 was detected at 6 h following reperfusion and peaked at 24 h both in cortical and basal ganglia regions, c-fos was markedly expressed at 1 h and the level peaked at 6 h in the ischemic hemispheres, and then reduced gradually. The TUNEL positive cells were markedly ohserved at 6 h. After treatment with Ecr, the positive reaction cells both of HSP70 and c-fos were significantly increased （ all P 〈0.01 ） and TUNEL positive cells were obviously reduced. Conclusion HSP70 and c-fos may be involved in pathophysiologic process of cerebral ischemia. Ecr may play neuroprotective roles for ischemical reperfusion injury.

关 键 词：脑缺血 热休克蛋白70 C-FOS 柘树制剂 

分 类 号：R96[医药卫生—药理学] R743[医药卫生—药学]