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重组腺病毒Ad-shRNA-NgR在大鼠缺血脑组织的转染效率研究 被引量：2

Transfection efficiency of recombinant adenovirus Ad-shRNA-NgR in ischemic brain tissues of rats
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摘要 目的探索用于RNA干扰的重组腺病毒Ad-shRNA-NgR转染大鼠缺血脑组织的适宜滴度。方法将低、中、高3种不同滴度(7.90×109、1.58×1010、3.16×1010pfu/ml)的重组腺病毒通过立体定位手术注射到大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)/再灌注模型大鼠(n=10)脑缺血区,于术后48 h和1周在荧光显微镜下观察各组大鼠脑组织中绿色荧光蛋白(green fluorescent protein,GFP)的表达强度并估算转染效率,通过HE染色检测炎性反应。结果中滴度和高滴度注射组大鼠脑组织腺病毒转染效率相似,为60%～70%,明显高于低滴度注射组(P<0.01)。在各组脑组织切片中,仅高滴度注射组有明显炎症反应,血管周围可见淋巴和巨噬细胞浸润。结论中等滴度(1.58×1010pfu/ml)的腺病毒能高效、稳定、低毒地转染大鼠缺血脑组织,推荐用于脑缺血基因干预治疗的实验研究。 Objective To explore a proper titer of RNA interference recombinant adenovirus Ad-shRNA-NgR transfecting ischemic region in rat brains for further study on gene therapy in cerebral ischemia.Methods Three different titers（7.90×10^9,1.58×10^10,and 3.16×10^10 pfu/ml） of recombinant adenovirus were respectively injected into the ischemic region of middle cerebral artery occlusion（MCAO）/reperfusion rats（n=10） by stereotactic surgery.Green fluorescent protein（GFP） expression in each group was observed to measure transfection efficiency of recombinant adenovirus in postoperative 48 h and 1 week.Inflammation was observed by HE staining.Results Transfection efficiencies of recombinant adenovirus in high and medium titer injection group were similar,about 60% to 70%,both significantly higher than low titer group（P〈0.01）.Besides,inflammation as perivascular infiltration of lymphoid and macrophage was only observed in high titer group.Conclusion Medium titer（1.58×10^10 pfu/ml） of recombinant adenovirus can stably,with high efficiency and low toxicity,transfect ischemic brain tissue in rats.
作者 吴小慧 秦新月 王敬 张沁丽 李鑫
机构地区 重庆医科大学附属第一医院神经内科
出处 《第三军医大学学报》 CAS CSCD 北大核心 2010年第8期802-804,共3页 Journal of Third Military Medical University
基金 国家自然科学基金(30470607)~~
关键词 重组腺病毒 Ad-shRNA-NgR 适宜滴度 转染 生物学标记 药理学 大鼠 recombinant adenovirus Ad-shRNA-NgR proper titer transfection biomarkers pharmacological rats
分类号 R394.33 [医药卫生—医学遗传学] R394.2 [医药卫生—医学遗传学]
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参考文献15

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共引文献3

同被引文献21

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引证文献2

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第三军医大学学报
2010年 第8期

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