GFP标记的重组腺病毒GDNF基因在脑缺血大鼠脑内的表达

THE EXPRESSION OF GFP-LABELED RECOMBINANT ADENOVIRUS GDNF GENE IN BRAIN OF RATS WITH CEREBRAL ISCHEMIA

目的:研究绿色荧光蛋白(GFP)标记的重组腺病毒胶质细胞源性神经营养因子(GDNF)基因在脑缺血大鼠脑内的表达。方法:采用改良的线栓法制作脑缺血再灌注模型,3d后用脑立体定位仪经侧脑室注射生理盐水或GDNF重组腺病毒液,注射前及注射后4d对大鼠进行神经功能损伤程度评分(NSS)。注射后4d麻醉处死大鼠,脑组织石蜡包埋。抗GFP免疫组织化学方法观察重组腺病毒GDNF基因在脑内的表达。结果:注射后4d,GDNF组与NS组相比,NSS评分显著降低;GDNF组,注射后4d,缺血侧纹状体内和梗死灶周围有大量GFP阳性细胞。结论:腺病毒介导的GDNF基因在脑缺血大鼠脑内能直接感染病灶周围神经细胞,表达GDNF蛋白,改善缺血后大鼠的神经功能状况。

Objective：To investigate the expression of GFP-labeled recombinant adenovirus GDNF gene in rat brain after focal cerebral ischemic reperfusion.Methods：The model of middle cerebral artery occlusion（MCAO） and reperfusion were set up and saline or solution of recombinant adenovirus GDNF gene were injected into the lateral cerebral ventricle of MACO model after reperfusion 3 days by stereotaxis instrument.Neurological severity score（NSS） tests were performed in all groups before injection and 4d after injection.Rats were anes-thetized and sacrificed 4d after injection.Anti-GFP immunohistochcmistry was used to check the expression of recombinant adenovirus GDNF gene.Results：Neurological Severity Score in GDNF group was significantly lower compared with NS group 4d after injection.In GDNF group,a large number of GFP positive cells were observed at the striatum in ischemic side and around infarction 4d after injection.Conclusion：Adenovirus-mediated GDNF gene could directly infect neural cells around lesion in the brain of rats with cerebral ischemia,expressed GDNF protein and improved neurological function.