化学合成小干扰RNA对大鼠脑内AQP4蛋白沉默效果验证与筛选  

Test and Selection of Effect of Silencing AQP4 Gene with Chemical Synthesis siRNA

作　　者：宋振全[1] 赵旭[1,2] 刘恩智[1] 柳云恩 张兴[2] 张海松[1] 单提坤 赵航[2] 

机构地区：[1]沈阳军区总医院神经外科,辽宁沈阳110016 [2]辽宁医学院研究生学院,辽宁锦州121001 [3]沈阳军区重症( 战) 创伤实验室,辽宁沈阳110016

出　　处：《创伤与急危重病医学》2014年第2期73-76,共4页Trauma and Critical Care Medicine

基　　金：辽宁省科技攻关课题(2009225018)

摘　　要：目的针对大鼠脑内AQP4蛋白使用化学合成法制备数条小干扰RNA链(siRNA),对其中两条链进行沉默效果验证,并筛选出效果显著的小干扰RNA链。方法根据大鼠的AQP4mRNA序列选择2个不同的靶点,采用化学合成的方法分别合成2条siRNA,并同时合成阴性对照siRNA。雄性成年Wistar大鼠18只,随机分为注射阴性对照液组、注射725链组和注射1006链组,并分别经侧脑室注射相应药物。6 h后将其全部处死,利用免疫组织化学法和原位杂交法分别检测脑内AQP4蛋白和mRNA的表达水平。结果给药6 h后,注射725链siRNA组大鼠脑内AQP4在蛋白质和基因两个水平均显著低于注射阴性对照液组和注射1006链组(P<0.05)。结论脑室注射725链siRNA可以在6 h内有效沉默大鼠脑内AQP4的表达。Objective Prepared several small interfering RNAs （siRNA）for AQP4 protein in rat brain by chemical synthe-sis.The silencing effect of two chains were verified and selected the one which was more effective.Methods Select two dif-ferent target sequences of the rat according to the AQP4 mRNA.Two siRNAs were synthesised by chemical synthesis,mean-while negative siRNA was synthesised as control.18 male adult Wistar rats were randomly divided into negative siRNA in-jection group,725 line injection group and 1006 line injection group.Corresponding drugs were injected into the left ventri-cle respectively.All animals were executed after 6 hours.The protein expression of AQP4 was detected by immunohisto-chemical staining and the AQP4mRNA expression was detected by in situ hybridization.Results 6 hours after injection,the expression of AQP4 protein and AQP4 mRNA of brain in the 725 line injection group were significantly lower than those of the 1006 line injection group and the negative siRNA injection group （P＆lt;0.05）.Conclusion 725 line injection could ef-fectively silence the expression of AQP4 in rat brain in 6 hours.

关 键 词：RNA干扰 AQP4蛋白 化学合成 沉默效果 aquaporin4 

分 类 号：R393[医药卫生—基础医学]