ZFP580对大鼠心肌缺血/再灌注后心室重塑的影响

Effects of ZFP580 on ventricular remodeling after myocardial ischemia/reperfusion in rats

目的:探究锌指转录因子(ZFP580)与心肌缺血/再灌注损伤后心室重塑的关系。方法:72只SD大鼠随机分为假手术(sham)组(n=8)和心肌缺血/再灌注(I/R)组(n=64),其中I/R组分别在再灌注后的0.5 h、1 h、2h、4 h、1 d,7 d,14 d,28 d处死后取材,观察心肌组织中ZFP580的表达。培养大鼠H9C2心肌细胞,每组设3个复孔,分别在转化生长因子β1(TGF-β1)刺激0 h、8 h、16 h、24 h后观察心肌细胞肥大情况,并检测心肌细胞中β-MHC、心房利钠肽(ANP)以及ZFP580 m RNA的表达。利用慢病毒介导的基因转染获得高表达ZFP580的H9C2心肌细胞,转染72h后,检测心肌细胞中基质金属蛋白酶3(MMP-3)的表达。结果:成功建立心肌缺血/再灌注损伤模型,大鼠心肌I/R损伤后第14天,心肌组织大面积梗死,心肌细胞呈嗜酸性变。大鼠心肌组织中ZFP580及TGF-β1表达上调。TGF-β1(5 ng/ml)刺激H9C2心肌细胞后诱导心肌细胞肥大,心肌细胞肥大标志蛋白β-MHC、ANP表达上调,且心肌细胞中ZFP580m RNA表达上调(P<0.05)。高表达ZFP580的H9c2心肌细胞中MMP-3表达下调(P<0.05)。结论:锌指转录因子ZFP580可能参与了心肌缺血/再灌注后心室重塑的过程,其作用可能与参与TGF-β1诱导的心肌细胞肥大过程以及抑制心肌细胞产生MMP-3有关。

Objective： To investigate the relationship between zinc finger protein（ZFP580）and ventricular remodeling after myocardial is- chemia/reperfusion（I/R） injury in rats. Methods： Seventy-two rats were divided into sham group and I/R groups which would be tested in se-lies time of 0.5 h, 1 h, 2 h, 4 h, 1 d,7 d,14 d,28 d after reperfusion to observe the expression of ZFP580 in rat myocardium. The H9C2 ceils were cultured and treated with transforming growth faetor-betal （TGF-β1） to establish cardiac hypertrophy in vitro model in series time of 0 h, 8h, 16 h and 24 h. The cardiornyocyte hy-pertrophy morphology was measured. The mRNA levels of atrial natriuretic peptide（ANP）, myosin heavey chain beta（β-MHC） and ZFP580 genes were quantified. The protein levels of MMP-3 and ZFP580 were quantified after H9C2 ceils were transfected hy lentiviral-mediated ZFP580 gene. Results： Myocardial I/R injury model was successfully established. Myocardial tissue in rats had large area infarction, and myocardial cells were eosinophilic changed. The increased level of ZH：＇580 protein was observed in the cardiomyocytes around infarction zone. The expression of TGF-β1 in myocardium was up-regulated ＇after myocardial I/R injury. TGF-β1 （5 ng/ml） treatment could induce cardiomyocyte hypertrophy in H9C2 cells. TGF-β1 treatment increased the cell size and mRNA levels of ANP and β-MHC genes （P 〈 0.05）, which represent degree of cardiac hypertrophy. TGF-β1 treatment also increased the protein levels of ZFP580 in H9C2 cells （ P 〈 0.05）. In the H9C2 cells transfected by lentiviral-mediated gene, the protein level of MMP3 was decreased （ P 〈 0.05）. Conclusion： ZFP580 is probably related with ventricular remodeling after myocardial I/R injury by involving TGF-β1 induced cardiomyocyte hypertrophy and attenuating MMP-3 production.