RNA干扰技术抑制可溶性环氧化物水解酶对急性心肌缺血坏死的影响  

Influence of soluble epoxide hydrolase inhibited by RNA interference on acute myocardial ischemic necrosis

作　　者：杜广胜[1] 徐雪晶[1] 贺莉[1] 马业新[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院心内科,武汉430030

出　　处：《中国急救医学》2011年第9期815-818,I0002,共5页Chinese Journal of Critical Care Medicine

摘　　要：目的应用RNA干扰（RNAinterference，RNAi）技术下调可溶性环氧化物水解酶（solubleEpoxideHydrolase，sEH）的表达，观察其对异丙肾上腺素（isoproterenol，ISO）诱导的急性缺血小鼠心肌的凋亡相关基因和心功能的影响。方法采用ISO注射液20mg／（kg·d）连续腹腔注射3d，建立小鼠急性心肌缺血模型，分为正常对照组（C组）、心肌缺血组（MI组）、缺血加阴性质粒组（PCN组）和缺血加EH-2质粒组（EH-2组）。利用构建好的靶向sEH基因的特异性小干扰RNA质粒EH-2，下调小鼠sEH基因的表达，用心脏超声观察各组左室舒张末期内径（LVEDd）、左室射血分数（EF）、左室短轴缩短率（FS）等变化。Westernblotting法检测各组Bcl-2、Bax和sEH的表达变化。结果与对照组相比，心肌缺血组EF和Fs值降低，LVEDd增加；心肌Bax表达升高，而Bcl-2表达降低（P〈0．01）。而缺血加EH-2质粒组sEH的表达降低，并且与MI组和PCN组相比，EF和Fs值增加，LVEDd减小；心肌Bax表达降低，Bcl-2表达升高（P〈0．01）。结论用RNA干扰技术可下调小鼠体内sEH的表达，从而增加抑凋亡基因Bcl-2的表达，改善ISO诱导的急性心肌缺血和心功能不全。Objective To investigate the effects of RNA interference （RNAi） downregutating soluble epoxide hydrolase （sEH） gene on apoptosis - related genes and cardiac function of mice with isoproterenol （ISO） induced myocardial ischemic necrosis. Methods A myocardial ischemic necrosis model of mice was established by abdominal cavity injection of [SO 20 mg/（ kg. d） for three days. The rats were divided into four groups： control group （C group） , myocardial ischemic necrosis group （MI group） and myocardial ischemia treated with negative plasmid group （PCN group）, myocardial ischemia treated with EH - 2 plasmid group （ EH - 2 group ）. Inhibition of sEH by EH - 2 plasmid were constructed the siRNA expression vectors specific to sEH gene. Left ventricular end diastolic diameters （LVEDd）, ejection fraction （EF）, fractional shortening （FS） of four groups were measured by echocardiography. The protein expression levels of Bcl -2, Bax and sEH were detected by Western blotting. Results Compared with C group, EF and FS decreased, LVEDd increased in myocardial ischemic necrosis groups. The expression levels of Bax increased, and the expression levels of Bcl -2 decreased （P 〈 0. 01 ）. However EH -2 group compare with MI group and PCN group, EF and FS increased, LVEDd decreased. The expression levels of sEH and Bax decreased, and the expression levels of Bcl - 2 increased （ P 〈 0.01 ）. Conclusion In vivo, RNAi can inhibit the expression of sEH, increase the expression of antiapoptotic gene, such as Bcl - 2, and ameliorate heart failure and myocardial ischemic injury induced by ISO.

关 键 词：异丙肾上腺素 可溶性环氧化物水解酶 RNA干扰 

分 类 号：R-332[医药卫生]