Malhotra A, Reich D, Reich D, Nakouzi A, Sanghi V, Geenen D L, Buttrick P M
Division of Cardiology, Albert Einstein College of Medicine, Bronx, NY, USA.
Circ Res. 1997 Dec;81(6):1027-33. doi: 10.1161/01.res.81.6.1027.
A cardiomyopathy that is characterized by an impairment in diastolic relaxation and a loss of calcium sensitivity of the isolated myofibril has been described in chronic diabetic animals and humans. To explore a possible role for protein kinase C (PKC)-mediated phosphorylation of myofibrillar proteins in this process, we characterized the subcellular distribution of the major PKC isoforms seen in the adult heart in cardiocytes isolated from diabetic rats and determined patterns of phosphorylation of the major regulatory proteins, including troponin I (TnI). Rats were made diabetic with a single injection of streptozotocin, and myocardiocytes were isolated and studied 3 to 4 weeks later. In nondiabetic animals, 76% of the PKC epsilon isoform was located in the cytosol and 24% was particulate, whereas in diabetic animals, 55% was cytosolic and 45% was particulate (P < .05). PKC delta, the other major PKC isoform seen in adult cardiocytes, did not show a change in subcellular localization. In parallel, TnI phosphorylation was increased 5-fold in cardiocytes isolated from the hearts of diabetic animals relative to control animals (P < .01). The change in PKC epsilon distribution and in TnI phosphorylation in diabetic animals was completely prevented by rendering the animals euglycemic with insulin or by concomitant treatment with a specific angiotensin II type-1 receptor (AT1) antagonist. Since PKC phosphorylation of TnI has been associated with a loss of calcium sensitivity of intact myofibrils, these data suggest that angiotensin II receptor-mediated activation of PKC may play a role in the contractile dysfunction seen in chronic diabetes.
在慢性糖尿病动物和人类中,已发现一种以舒张期松弛受损和离体肌原纤维钙敏感性丧失为特征的心肌病。为了探究蛋白激酶C(PKC)介导的肌原纤维蛋白磷酸化在此过程中可能发挥的作用,我们对从糖尿病大鼠分离的心肌细胞中所见的成年心脏主要PKC亚型的亚细胞分布进行了表征,并确定了包括肌钙蛋白I(TnI)在内的主要调节蛋白的磷酸化模式。大鼠通过单次注射链脲佐菌素诱导糖尿病,3至4周后分离并研究心肌细胞。在非糖尿病动物中,76%的PKCε亚型位于细胞质中,24%为颗粒状;而在糖尿病动物中,55%为细胞质型,45%为颗粒状(P <.05)。成年心肌细胞中所见的另一种主要PKC亚型PKCδ的亚细胞定位未显示变化。同时,与对照动物相比,从糖尿病动物心脏分离的心肌细胞中TnI磷酸化增加了5倍(P <.01)。通过用胰岛素使动物血糖正常化或同时用特异性血管紧张素II 1型受体(AT1)拮抗剂治疗,可完全防止糖尿病动物中PKCε分布和TnI磷酸化的变化。由于TnI的PKC磷酸化与完整肌原纤维的钙敏感性丧失有关,这些数据表明血管紧张素II受体介导的PKC激活可能在慢性糖尿病所见的收缩功能障碍中起作用。