利用RNA干扰技术治疗慢性心衰大鼠的研究  

The Research of RNA Interference Therapy for Chronic Heart Failure Rats

作　　者：杨广[1] 李小鹰[1] 鲁晓春[1] 

机构地区：[1]解放军总医院老年心内科

出　　处：《中国分子心脏病学杂志》2007年第2期99-103,共5页Molecular Cardiology of China

基　　金：国家973项目资助(G2000056906)

摘　　要：目的利用RNA干扰(RNA interference,RNAi)技术抑制心衰大鼠受磷蛋白(phospho- lamban,PLN)mRNA的表达,改善心脏功能。方法构建表达PLN shRNA的重组腺相关病毒载体(rAAV2-phRi 1及rAAV2-phRi 2);通过经腹心包穿刺术将rAAV2-phRi1及rAAV2-phRi2注入心衰大鼠心包内,基因导入后10 d,30 dRT-PCR法检测RNAi对心肌细胞PLN mRNA的抑制效应,并测定肌浆网Ca^(2+)-ATP酶(sarcoplasmie retieulum Ca^(2+)ATPase,SERCA2a)活性,比较血流动力学的变化。结果rAAV2-phRil及rAAV2-phRi2心包注射后,与心衰组比较,10 d时PLN mRNA分别降低18．76%/ 16．36%,SERCA2a活性及血流动力学指标无明显改善;30 d时rKAV2-phRi1及rAAV2-phRi2对PLN mRNA的抑制效率为85．32%/67．65%,SERCA2a活性增加104%/74%,血流动力学显著改善(P＜0．05),达到假手术组水平(P＞0．05);rAAV2-phRi1的抑制效率,对心功能的改善显著优于rAAV2- phRi2(P＜0．05)。结论在体水平上,RNAi能够抑制心衰大鼠PLN mRNA表达,改善心脏功能, rAAV2-phRi1优于rAAV2-phRi2。Objective To inhibit phospholamban （PLN） mRNA expression and improve cardiac function of heart failure （HF） rats by RNA interference （RNAi）. Methods Construct recombinant adeno-asociated virus vector2 expressing PLN shRNA（ rAAV2-phRil and rAAV2-phRi 2） ; rAAV2 was injected into HF rats by abdominal pericardia intracavitary injection. After 10 and 30 days, Inhibitory effects of RNAi on PLN mRNA were detected by RT-PCR, sarcoplasmic reticulum Ca^2＋ -ATPase＇ activity （SERCA2a） and hemodynamic indexes were measured. Results Compare to HF rats, at 10 days, rAAV2-phRi1 and rAAV2-phRi2 decreased respectively PLN mRNA by 18.76/16.36% and SERCA2a activity, hemodynamic indexes had no significant（P 〉0.05） ;At 30 days, rAAV2-phRi1 and rAAV2-phRi2 decreased PLN mRNA respectively by 85.32/67.65%, SERCA2a activity increased by 104%/74% and hemodynamic indexes improved significantly than those of HF rats （ P 〈 0.05 ） ; rAAV2-phRi1 was superior to rAAV2-phRi2 （ P 〈 0.05） and reached control group level（P 〉 0.05）. Conclusion In vivo, RNAi can inhibit the expression of PLN mRNA, improved cardiac function; rAAV2-phRil was superior to rAAV2-phRi2.

关 键 词：RNA干扰 受磷蛋白 肌浆网CA^2+-ATP酶 心衰 大鼠 

分 类 号：R541.6[医药卫生—心血管疾病]