Increased basal contractility of cardiomyocytes overexpressing protein kinase Cε and blunted positive inotropic response to endothelin-1
Increased basal contractility of cardiomyocytes overexpressing protein kinase Cε and blunted positive inotropic response to endothelin-1
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DOI：
10.1016/s0008-6363(01)00225-5
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发表时间：
2001-06-01
影响因子：
10.8
通讯作者：
Prestle, J
中科院分区：
文献类型：
--
作者：
Baudet, S;Weisser, J;Prestle, J
目的：蛋白激酶C （PKC）被认为通过血管活性肽如内皮素-1 （ET-1）参与调节哺乳动物心脏兴奋-收缩耦联过程。然而，特定PKC异构体的激活与ET-1介导的收缩性增加之间的因果关系仍然是推断性的。方法：通过腺病毒介导的基因转移，在培养的成年兔心室肌细胞中特异性过表达PKC epsilon （Ad-PKC epsilon）。在Ad-PKC epsilon和Ad-LacZ对照转染的细胞中，测量了基础和et -1刺激条件下的肌细胞缩短和[Ca2+](i)瞬变。结果：Ad-PKC epsilon感染导致PKC epsilon蛋白水平强烈的病毒剂量依赖性增加，而其他PKC亚型的蛋白表达保持不变。使用100个斑块形成单位/肌细胞的多重感染，与对照组相比，基础和辅助因子依赖性PKC ε激酶活性分别增加了28倍和90倍。心肌细胞基底部分缩短和[Ca2+](i)，瞬态振幅均增加21% (P
Objective: Protein kinase C (PKC) is thought to be involved in the regulation of the mammalian cardiac excitation-contraction coupling process by vasoactive peptides Like endothelin-1 (ET-1). However, the demonstration of a causal link between activation of specific PKC isoforms and the increase in contractility mediated by ET-1 is still inferential. Methods: By means of adenovirus-mediated gene transfer, we specifically overexpressed PKC epsilon in cultured adult rabbit Ventricular myocytes (Ad-PKC epsilon). Myocyte shortening and [Ca2+](i) transients under basal and ET-1-stimulated conditions were measured in Ad-PKC epsilon and Ad-LacZ control transfected cells. Results: Infection with Ad-PKC epsilon resulted in a strong, virus dose-dependent increase in PKC epsilon protein Levels, whereas protein expression of other PKC isoforms remained unchanged. Using a multiplicity of infection of 100 plaque-forming units/myocyte, basal and cofactor-dependent PKC epsilon kinase activity was increased 28- and 90-fold, respectively, when compared to control. Myocyte basal fractional shortening and [Ca2+](i), transient amplitude were both increased by 21% (P