磷酸受纳蛋白反义RNA转染减轻心肌梗死后心室重塑和心力衰竭的发生  

Phospholamban antisense RNA transfer attenuates post-infarction remodeling and preserves cardiac functions

作　　者：汪国忠[1] 吕树铮[1] 柳景华[1] 郭成军[1] 周渊[1] 苑飞[1] 宋现涛[1] 郭长江[1] 金泽宁[1] 

机构地区：[1]首都医科大学附属北京安贞医院心内科 北京市心肺血管疾病研究所,100029

出　　处：《中华医学杂志》2010年第20期1389-1394,共6页National Medical Journal of China

基　　金：北京市科委重大攻关项目（D090600600091）;中国博士后基金（20080430441）

摘　　要：目的 探讨磷酸受纳蛋白反义RNA真核表达质粒转染心肌对心肌梗死后心脏功能及病理性心室重塑的影响.方法 Wistar大鼠70只,分为生理盐水组（28只）,假手术组（13只）,冠状动脉结扎制备大鼠心肌梗死后心力衰竭模型,直接心肌注射转染磷酸受纳蛋白反义RNA真核表达质粒（PcDNA4-asPLB组,29只）,评价假手术组、生理盐水组及PcDNA4-asPLB组心脏舒缩功能及血流动力学改变.检测各组大鼠心肌磷酸受纳蛋白（PLB）、肌浆网Ca^2＋ATP酶（SERCA2a）蛋白表达水平及心房尿钠肽（ANP）及脑型尿钠肽（BNP）mRNA表达;病理检查分析各组大鼠心肌梗死面积、前壁变薄比、心肌细胞横径及纤维沉积状况.结果 超声心动图及血流动力学检测发现心肌梗死后PcDNA4-asPLB组大鼠心脏舒缩功能及血流动力学明显优于生理盐水组,尤以心肌梗死后第4周更为明显[左室射血分数（39.4±7.8）%比（30.9±7.4）%,P〈0.05;压力变化的最大值（1545±127）mm Hg·s^-1比（1172±91）mm Hg·s^-1,P〈0.05].PcDNA4-asPLB组PLB表达明显受抑,仅为生理盐水组的50%（PLB/β-肌动蛋白比值,0.28±0.07比0.57±0.11,P〈0.05）,SERCA2a表达较生理盐水组增强[（1.47±0.21）μmol·min^-1·g^-1蛋白比（0.34±0.13）μmol·min^-1·g^-1蛋白,P〈0.05].生理盐水组前壁变薄比、心肌细胞横径、纤维沉积及胚胎基因表达等心室重塑改变较PcDNA4-asPLB组明显.结论 PcDNA4-asPLB转染抑制PLB表达,减轻和逆转心肌梗死后心室重塑,延缓心肌梗死后心力衰竭的发生.Objective To investigate whether the gene transfer of phospholamban antisense RNA could inhibit remodeling and preserve cardiac function after myocardial infarction. Methods Wistar rats received a ligation of left coronary with a direct intramyocardial injection of phospholamban antisense RNA eukaryon vector PcDNA4-asPLB. The cardiac function, hemodynamics and ventricular geometry of three groups （shame, saline injection and PcDNA4-asPLB injection） were studied by echocardiography and left ventricle hemodynamic recording. The levels of phospholamban （PLB） and sarcoplasmic reticulum Ca^2＋-ATPase （SERCA2a） were analyzed by Western blot and the expressions of atrial natriuretic peptide （ANP） and brain natriuretic peptide （BNP） examined by RT-PCR. The histological study was performed to evaluate the collage content and cardiomyocyte fiber size. Results The PcDNA4-asPLB injection group had significantly better systolic cardiac function and diastolic function [LVEF（39. 4±7. 8）% vs （30. 9±7.4）%, P〈0.05; dp/dt Max （1545 ±127） mm Hg·s^-1 vs （1172±91） mm Hg·g^-1, P〈0.05）]. Compared with saline injection, the PLB expression was inhibited by 50% in PcDNA4-asPLB injection group （PLB/β-actin ratio, 0. 28±0. 07 vs 0. 57±0. 11, P〈0. 05） and the function of SERCA2a was enhanced [（1.47±0.21） μmol·min^-1·g^-1 protein vs （0.34±0.13）μmol·min^-1·g^-1 protein, P〈0.05]. The expressions of ANP and BNP in the saline injection group were elevated as compared to those in the PcDNA4-asPLB injection group. Histological study also showed that the collage density and thecardiomyocyte fiber size in the saline injection group were worse than those in the PcDNA4-asPLB injection group.Conclusion Intramyocardial injection of phospholamban antisense RNA eukaryon vector PcDNA4-asPLB after myocardial infarction results in PLB expression inhibition.attenuates ventricular remodeling and improves systolic and diastolic cardiac functions.

关 键 词：心肌梗死 心力衰竭 充血性 RNA 反义 磷酸受纳蛋白 

分 类 号：R541.6[医药卫生—心血管疾病]