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[兔收缩性和舒张性心力衰竭模型中心脏功能及心肌钙调节蛋白表达与活性的比较。]

[Comparison of cardiac function and expression and activity of myocardial calcium regulatory proteins in rabbit systolic and diastolic heart failure models.].

作者信息

Wang Lei, Zhang Shi-Jie, Wang Hai-Peng, Zou Cao, Liu Zhi-Hua

机构信息

Department of Cardiology, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.

出版信息

Sheng Li Xue Bao. 2009 Dec 25;61(6):551-8.

Abstract

The aim of the present study is to investigate the differences in cardiac function, and the expression and activity of calcium regulatory proteins between rabbit systolic heart failure (SHF) and diastolic heart failure (DHF) models. New Zealand white rabbits were randomly divided into three groups: sham operation (SO) group, DHF group (receiving abdominal aortic constriction) and SHF group (receiving aortic valve destruction and abdominal aortic constriction). The cardiac function was detected by echocardiographic and hemodynamic assays. The mRNA expression levels of sarcoplasmic reticulum Ca(2+) ATPase 2a (SERCA2a) and phospholamban (PLB) were evaluated by RT-PCR. The protein expression levels of SERCA2a, PLB, phosphoserine 16-PLB (pSer-16-PLB) and protein kinase A (PKA) were evaluated by Western blot, and the phosphorylation status of PLB was determined by the ratio of pSer-16-PLB protein level to that of PLB. The activity of SERCA2a was measured through inorganic phosphate. The activity of PKA was measured by gamma-(32)P ATP-binding assays. Compared with SO group, there were significantly increased ventricular wall thickness, raised left ventricular end diastolic pressure (LVEDP), reduced diastolic function in DHF group (P<0.05 or P<0.01), and significantly increased ventricular cavity size and LVEDP, reduced systolic function in SHF group (P<0.05 or P<0.01). The expression levels of SERCA2a in DHF and SHF groups were lower than that in SO group (P<0.05), while the expression and activity of PKA in DHF and SHF groups were higher than that in SO group (P<0.05 or P<0.01), and there was no significant difference between DHF and SHF groups. The expression levels of PLB and pSer-16-PLB as well as the phosphorylation status of PLB and activity of SERCA2a in SHF group were lower than those in DHF and SO groups respectively. Posing a contrast, the phosphorylation status of PLB and activity of SERCA2a in DHF group were higher than that in SO group (P<0.05). These results indicate that the SHF and DHF models were successfully established, and there are some differences in the expression and activity of calcium regulatory proteins between two models.

摘要

本研究旨在探讨兔收缩性心力衰竭(SHF)和舒张性心力衰竭(DHF)模型之间心脏功能以及钙调节蛋白表达和活性的差异。将新西兰白兔随机分为三组:假手术(SO)组、DHF组(接受腹主动脉缩窄)和SHF组(接受主动脉瓣破坏及腹主动脉缩窄)。通过超声心动图和血流动力学检测来检测心脏功能。采用逆转录聚合酶链反应(RT-PCR)评估肌浆网Ca(2+)ATP酶2a(SERCA2a)和受磷蛋白(PLB)的mRNA表达水平。采用蛋白质免疫印迹法评估SERCA2a、PLB、磷酸化丝氨酸16-PLB(pSer-16-PLB)和蛋白激酶A(PKA)的蛋白表达水平,并通过pSer-16-PLB蛋白水平与PLB蛋白水平的比值来确定PLB的磷酸化状态。通过无机磷酸盐测定SERCA2a的活性。采用γ-(32)P ATP结合分析法测定PKA的活性。与SO组相比,DHF组心室壁厚度显著增加、左心室舒张末期压力(LVEDP)升高、舒张功能降低(P<0.05或P<0.01),SHF组心室腔大小和LVEDP显著增加、收缩功能降低(P<0.05或P<0.01)。DHF组和SHF组SERCA2a的表达水平低于SO组(P<0.05),而DHF组和SHF组PKA的表达和活性高于SO组(P<0.05或P<0.01),DHF组和SHF组之间无显著差异。SHF组PLB和pSer-16-PLB的表达水平以及PLB的磷酸化状态和SERCA2a的活性分别低于DHF组和SO组。相反,DHF组PLB的磷酸化状态和SERCA2a的活性高于SO组(P<0.05)。这些结果表明成功建立了SHF和DHF模型,且两种模型之间钙调节蛋白的表达和活性存在一些差异。

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