成年大鼠侧脑室注射与脑实质注射腺病毒表达载体转染脑细胞的比较研究

目的比较观察脑室注射和脑实质注射腺病毒表达载体转染脑细胞的范围及表达产物的程度。方法分别将Ad5CMV LacZ表达载体（20μl,病毒滴度为1．01×10^5pfu／ml）注入成年SD大鼠右侧脑室、尾壳核、内侧隔核-斜角带复合体和背侧海马,在注射后3～28d将大鼠脑制作成为切片,进行X-gal组织化学反应,观察被转染和表达的细胞形态和数目。结果在侧脑室内注射后3d,右侧侧脑室室管膜细胞及其下层开始有病毒载体转染和表达,继之向对侧侧脑室、第三脑室和第四脑室转染和表达产物,脊髓中央管壁偶尔有被转染和表达的细胞;而将等体积和等滴度的AdSCMV LacZ载体注入尾壳核、内侧隔核-斜角带复合体和背侧海马,则均出现直径约2～5mm的转染范围,表达产物的细胞离注射原位最远处约为5—6mm,细胞表达产物很局限并相对集中;而侧脑室注射病毒载体可以随脑脊液流动而使转染范围广,但被转染细胞主要集中在室管膜及其下层,离脑室壁最远处也只在0．5—2mm范围内。结论侧脑室注射病毒表达载体便于基因治疗弥漫性脑疾病;而脑实质注射病毒表达载体则利于基因治疗局部性脑疾病。

A comparison investigation into the effects of adenoviral vector delivery to the brain between the lateral ventricle injection and cerebral parenchyma injection

Objective To compare the effects of transferring adenoviral vector carrying LacZ gene from the two different pathways ：lateral ventricular injection and cerebral parencbyma injection. Methods Injections of Ad5 CMV LacZ vector （20μl,1.01 × 10^5 pfu/ml） into the lateral ventricle, medial septal nucleus-diagonal complex, caudate-putmen and hippocampus were given, respectively. After 3 - 28 days of injection,the rats were sacrificed to detect the transfected and expressed cells in the above four brain areas by means of X-gal histoehemistry. Results Three days After infection, the cells from the lateral ventricle to the fourth ventricle, and to the spinal cord were transfeeted and expressed. 7 -28 days after injection ,the ceils in medial septal nucleus-diagonal complex ,hippocarnpus and eaudate-putman were transfected and expressed. Conclusion The above gene deliveries of adenoviral vector to the brain are potential pathways for transferring a target gene to treating brain diseases.