摘要
目的探讨RNA干扰(RNA interference,RNAi)对大鼠脑内水通道蛋白4(aquaporin4,AQP4)表达的抑制作用。方法构建特异性抑制AQP4表达的短发夹环质粒载体AQP4RNAi pGenesil-2。Wistar成年大鼠80只随机分为假手术组、脂质体组、对照质粒组和AQP4RNAi组(n=20),采用右侧脑室给药的方法分别于术后2、5、7、15d进行一般情况观察,对双侧大脑半球、脑干和小脑进行光、电镜检查,运用免疫荧光和原位杂交方法分别检测AQP4蛋白和mRNA的表达。结果脑室内注入AQP4短发夹环质粒载体后大鼠的精神、运动、行为未见明确异常,光、电镜病理检查发现神经元和胶质细胞形态、分布和脑室的形态结构与假手术组相比未见明显改变。AQP4蛋白和mRNA的表达呈进行性减少,至术后第7天,分别减少62.75%±3.6%和63.65%±2.35%。结论短发夹环AQP4RNAi质粒载体能有效减少脑内AQP4的表达,对中枢神经的功能无明显影响。
Objective To observe the expression of AQP4 in brain of rat after RNA interference. Methods We recombinated a short hairpin plasmid vector pGenesil-2 that can specifically suppress AQP4 expression. Eighty Wistar rats were divided into AQP4 RNA interference group, sham operation group, Lipefectamin group and contrast plasmid vector group. We injected recombinant plasmid pGenesil into right lateral cerebral ventricle with Lipofectamin^TM2000. AQP4, AQP4 mRNA were observed by immunohistochemistry analysis and semiquantitative in situ hybridazation experiments on 2, 5,7,15 days after transfection. Results Semiquantitative in situ hybridazation experiments and immunohistochemistry analysis showed that AQP4 silencing determined a progressive and parallel reduction in AQP4 mRNA and protein. Consciousness, behavior and limb movement was normal, and morph of neuron and astrocyte was not changed following AQP4 RNAi in brain of rat. Conclusions This result indicates that recombinant plasmid pC, enesil-2 can inhibite AQP4 expression of brain in vivo. It has no influence on function of CNS of rat.
出处
《中国神经精神疾病杂志》
CAS
CSCD
北大核心
2007年第12期719-723,共5页
Chinese Journal of Nervous and Mental Diseases
基金
军队"十五"指令性课题(编号:01L060)
