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血管紧张素Ⅱ1型受体亚型鉴别的RNAi应用研究

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摘要 目的针对小鼠血管紧张素Ⅱ 1型受体(Angiotensin Ⅱ Receptor 1,AT1R)两亚型之间无工具药区分的现状,研究应用RNA干扰(RNAinterference,RNAi)技术来区分两种亚型受体功能的可行性。方法选择雄性C57BL小鼠作为动物模型,通过在脑干孤束核(nucleus tractus solitarius,NTS)注射携带短发夹RNA(shor thairpin RNA,shRNA)腺病毒的方法,喂养10d后取出脑干组织做标本,使用原位杂交的方法检测每只小鼠脑干NTS部位AT1R mRNA两种亚型的表达情况,同时使用免疫组织化学的方法显示受体蛋白表达的情况。结果注入携带针对AT1aR mRNA的shRNA腺病毒后,AT1aRmRNA的表达降低了(61.2±6.8)%(P<0.01),AT1bR mRNA的表达则不受影响(P>0.05);改用AT1bR mRNA相应的shRNA腺病毒,AT1bRmRNA的表达降低了(51.6±5.2)%(P<0.01),AT1aR mRNA的表达基本不受影响(P>0.05)。用免疫组织化学显示AT1R蛋白表达情况,发现两种情况下表达均出现不同程度的下降。结论所设计的针对不同亚型AT1R的shRNA,可以特异地沉默各自不同受体的mRNA表达,并同时在蛋白水平上显示出AT1R受到抑制的结果,故可以将其应用于区别AT1R两种亚型受体功能的研究。
机构地区 广东省人民医院
出处 《广东医学》 CAS CSCD 北大核心 2008年第7期1117-1120,共4页 Guangdong Medical Journal
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