ACE siRNA对人脐静脉内皮细胞ET-1表达的影响

目的:观察小干扰RNA(siRNA)对人脐静脉内皮细胞CRL-1730血管紧张素转换酶(ACE)mRNA与蛋白表达的作用及对内皮素-1(ET-1)表达的影响。方法:设计3条针对人ACE基因的siRNA,脂质体转染CRL-1730细胞,RT-PCR检测ACEmRNA的表达,ELISA法检测ACE蛋白含量;并比较ACE siRNA与卡托普利(Captopril)对血管紧张素II同步处理6、12、24h后CRL-1730细胞ET-1的表达。结果:ACE siRNA能够抑制ACE mRNA及蛋白表达,以48h为甚(mRNA水平下调71.01%;蛋白水平下调68.13%,均P<0.01)。ACE siRNA和Captopril均能抑制CRL-1730细胞ET-1表达(P<0.05),ACE siRNA抑制作用优于Captopril(P<0.05)。结论:ACE siRNA能有效下调ACE的表达,并抑制血管紧张素II诱导的ET-1表达。

Effects of ACE siRNA on the Expression of Endothelin-1 in Human Umbilical Vein Endothelial Cells YANG

Objective To observe the effects of ACE siRNA on the expression of angiotensin converting enzyme mRNA in human umbilical vein endothelial cells CRL-1730 and the expression of endothelin-1. Methods Three siRNAs targeted against ACE gene were designed and synthesized, transfected into CRL-1730 with liposome,the expression of ACE mRNA and protein was detected with RT-PCR and ELISA, respectively. The expression of endothelin-1 stimulated with angiotensin I/and ACE siRNA or captopril at the time point of 6 h, 12 h and 24 h were compared. Results The expression of ACE mRNA and protein could inhibited with ACE siRNA,and reached the peak at 48 h（ mRNA decreased 71.01% ,P 〈 0.01 ; protein decreased 68.13% ）. ACE siRNA and captopril all could inhibit the expression of ET-1 in CRL-1730 cells（P 〈0.05） ,and the effect of ACE siRNA was better than that of captopril （ P 〈 0.05 ）. Conclusion RNA interference could effectively downregnlate the expression of ACE and inhibit the expression of ET-1 induced with angiotensin Ⅱ.