重组腺病毒介导Klotho基因转导对大鼠心力衰竭心肌重构的影响  

Impact of transfection with recombinant adenovirus vector-mediated Klotho gene on myocardial remodeling in a rat model of heart failure

作　　者：贾政[1] 魏玲[2] 刘茜[2] 朱姿英[2] 杨静[2] 杨晓华[2] 甘世保[2] 陈韦强 张丽萍[2] 

机构地区：[1]桂林医学院研究生学院,541004 [2]成都军区昆明总医院地方干部病房

出　　处：《中华心血管病杂志》2015年第3期219-226,共8页Chinese Journal of Cardiology

基　　金：成都军区昆明总医院院管课题（2012YG15）

摘　　要：目的 探讨重组腺病毒介导的Klotho基因转染对大鼠心力衰竭（心衰）进程以及心肌重构的影响,并探讨其机制.方法 SPF级SD大鼠,随机数表法分为5组,即正常对照组（对照组,未给予任何干预）,心衰组（只做心衰模型,不给予其他干预）,生理盐水组（在心衰模型的基础上尾静脉注射生理盐水,0.5 ml/只）,腺病毒载体介导的增强荧光蛋白组（Ad.EGFP组,在心衰模型的基础上用腺病毒载体介导增强荧光蛋白,2×1010 pfu,0.5 ml/只）,以及腺病毒载体介导的目的基因组（Ad.Klotho组,在心衰模型的基础上用腺病毒载体介导目的基因,2×1010pfu,0.5 ml/只）,每组5只.异丙肾上腺素连续腹腔注射法建立大鼠心衰模型.构建重组腺病毒载体pDC316-CMV-EGFP-rKlotho,标记为Ad.Klotho,并将其转导至相应的心衰大鼠模型.建模第6周,采用超声心动图测定各组大鼠的左心室射血分数（LVEF）;多道生理记录仪测定各组大鼠血液动力学变化;检测各组大鼠心肌组织病理学改变;冰冻切片后镜下观察各组大鼠心肌组织荧光蛋白表达水平;实时荧光定量聚合酶链反应评估各组大鼠心肌组织中Klotho基因的表达水平,以及心肌纤维化相关因子Ⅰ型胶原和Ⅲ型胶原的表达水平;ELISA法检测各组大鼠血浆中B型利钠肽（BNP）的表达水平.结果 心衰组、生理盐水组、Ad.EGFP组和Ad.Klotho组大鼠心肌组织苏木素伊红染色均可见心肌细胞断裂、心肌间质增生、心肌细胞肥大和结构改变.对照组大鼠心肌组织病理切片未见上述改变.通过比较发现,Ad.Klotho组大鼠心肌纤维化程度、重构程度以及心肌细胞肥大程度均较心衰组、生理盐水组和Ad.EGFP组低.建模第6周,Ad.Klotho组和Ad.EGFP组大鼠心肌组织中仍可见EGFP绿色荧光表达,而对照组、心衰组和生理盐水组则未见.各组大鼠心肌组织中Klotho、胶原Ⅰ a1和Ⅲa1 mRNA均有表达.其中,对照组和Ad.Klotho组KObjective To assess the impact of transfection with recombinant adenovirus vector-mediated Klotho gene on myocardial remodeling in a rat model of heart failure （HF） by intraperitoneal injection of isoproterenol.Methods Rats were divided into 5 groups by table of exponential random numbers：normal control group,HF group,saline-control HF group,recombinant adenovirus vector transfection group （Ad.EGFP group,2 × 1010 pfu,0.5 ml/rat）,pDC316-CMV-EGFP-rKlotho transfection group （Ad.Klotho group,n =5 each）.Left ventricular ejection fraction （LVEF） was obtained by echocardiography,hemodynamic parameters obtained by multi-channel physiological recorder,myocardial tissue underwent pathohistological examination.Additionally,the green fluorescin expression was observed on frozen heart section.Myocardial fibrosis correlated gene expression including Klotho gene,collagen Ⅰ and Ⅲ was detected by real time-PCR.Moreover,plasma levels of B-type natriuretic peptide （BNP） were measured with ELISA.Results Compared to saline control HF group,LVEF,LVSP and ± dp/dtmax were significantly increased,myocardial fibrosis and myocardial remodeling were significantly attenuated in the Ad.Klotho group and there was green fluorescin distribution in myocardial tissues of Ad.Klotho group.Klotho expression was down-regulated and collagen Ⅰ and Ⅲ expression was upregulated in HF rats compared to normal control group （all P ＜ 0.05） and these changes could be significantly reversed in Ad.Klotho group （all P ＜ 0.05）.Plasma BNP level was also significantly lower in Ad.Klotho group than in HF group （P ＜ 0.05）.Conclusions Klotho gene transfection could improve cardiac function and attenuate cardiac remodeling and reducing myocardial fibrosis.

关 键 词：心力衰竭 充血性 心室重构 KLOTHO基因 

分 类 号：R541.6[医药卫生—心血管疾病]