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心力衰竭家兔肌浆网钙回摄功能的异常及其影响因素

王蕾, 张世杰, 李莉, 兰超, 张瑞, 刘志华

郑州大学第一附属医院急诊科，郑州 450052；郑州大学第一附属医院检验科，郑州 450052；苏州大学附属第一医院心内科，苏州，215006

摘要

本文旨在探讨心力衰竭家兔心肌肌浆网钙回摄功能的异常及其影响因素和可能的机制。用主动脉破瓣术加腹主动脉缩窄术造心衰模型，用钙离子荧光成像仪检测心肌肌浆网钙回摄功能，用无机磷酸根法测定心肌肌浆网钙泵的活性，用Western blot检测各组心肌组织中肌浆网钙泵(SERCA2a)、受磷蛋白(PLB)、16位丝氨酸磷酸化-受磷蛋白(PLB-Ser16)、17位苏氨酸磷酸化-受磷蛋白(PLB-Thr17)、Ca2+∕钙调素依赖的蛋白激酶II (CaMKII)、蛋白激酶A (PKA)和蛋白磷脂酶(PP1α)的蛋白表达，用γ-32P底物掺入法测定CaMKII、PKA的活性。结果显示，与假手术组相比，心衰组钙回摄量显著下降(P < 0.01)，SERCA2a在心肌中的表达量和活性均显著下降(P < 0.05)，PLB的表达水平及其磷酸化位点(Ser16、Thr17)的磷酸化状态明显降低(P < 0.05)，且PKA和CaMKII的表达及活性均明显升高(P < 0.05或P < 0.01)，PP1α的表达量显著升高(P < 0.05)。以上结果证实心衰家兔模型心肌肌浆网钙回摄功能的异常与SERCA2a的表达活性、PLB及其磷酸化水平均显著下降有关，提示PLB的两个磷酸化位点(Ser16、Thr17)可能共同参与心衰家兔心肌肌浆网钙泵活性的调节。

关键词： 心力衰竭; 钙回摄; 肌浆网; 受磷蛋白

分类号：R541

[The abnormal calcium re-uptake from sarcoplasmic reticulum and influencing factors in failed rabbits.] [Article in Chinese]

Wang Lei, ZHANG Shi-Jie, Li Li, Lan Chao, Zhang Rui, LIU Zhi-Hua

Department of Emergency, the First Affiliated Hospital of Zhengzhou University, Zhengzhou,450052, China； Department of Clinical Laboratory, the First Affiliated Hospital of Zhengzhou University, Zhengzhou,450052, China； Department of Cardiology, the First Affiliated Hospital of Soochow University, Suzhou, 215006, China

Abstract

The aim of the present study was to investigate the abnormal calcium re-uptake function of myocardium sarcoplasmic reticulum (SR) in rabbits with heart failure, as well as potential mechanisms. Heart failure model was established in rabbits through aortic insufficiency and constriction of abdominal aorta. The SR Ca²⁺ re-uptake function was measured with a calcium imaging device. The activity of myocardium SR calcium adenodine triphosphatase 2a (SERCA2a) was measured by inorganic phosphate. The protein expressions of SERCA2a, CaMKII, PKA, PP1α, phospholamban (PLB), PLB-Ser¹⁶ and PLB-Thr¹⁷ were evaluated by Western blot. The activities of PKA and CaMKII were detected by γ-³²P substrate incorporation. The results showed that, compared with the sham operation group, the heart failure group exhibited reduced Ca²⁺ re-uptake amount (P < 0.01) and the expression and activity of SERCA2a (P < 0.05 or P < 0.01), decreased expression of PLB and its phosphorylation status in sites of Ser¹⁶ and Thr¹⁷ (P < 0.05), increased expressions and activities of PKA and CaMKII (P < 0.05 or P < 0.01), and increased expression of PP1α (P < 0.05). These results suggest that the abnormal Ca²⁺ re-uptake function in heart failure is related with reduced expression and activities of SERCA2a, as well as reduced expression of PLB and its phosphorylation status. Both PLB-Ser¹⁶ and -Thr¹⁷ may be involved in the regulation of myocardium SR calcium pump activity in heart failure.

Key words: Heart failure; calcium re-uptake; sarcoplasmic reticulum; phospholamban

收稿日期：2014-01-01　　录用日期：2014-04-03

通讯作者：张世杰　　E-mail: zsj03@zzu.edu.cn

引用本文：

王蕾, 张世杰, 李莉, 兰超, 张瑞, 刘志华. 心力衰竭家兔肌浆网钙回摄功能的异常及其影响因素[J]. 生理学报 2014; 66 (4): 483-488.

Wang Lei, ZHANG Shi-Jie, Li Li, Lan Chao, Zhang Rui, LIU Zhi-Hua. [The abnormal calcium re-uptake from sarcoplasmic reticulum and influencing factors in failed rabbits.] [Article in Chinese]. Acta Physiol Sin 2014; 66 (4): 483-488 (in Chinese with English abstract).