Centro de Investigaciones Cardiovasculares, Facultad de Ciencias Médicas, UNLP, 60 y 120 (1900) La Plata, Argentina.
J Physiol. 2010 May 1;588(Pt 9):1579-90. doi: 10.1113/jphysiol.2009.186619. Epub 2010 Mar 15.
Myocardial stretch elicits a biphasic contractile response: the Frank-Starling mechanism followed by the slow force response (SFR) or Anrep effect. In this study we hypothesized that the SFR depends on epidermal growth factor receptor (EGFR) transactivation after the myocardial stretch-induced angiotensin II (Ang II)/endothelin (ET) release. Experiments were performed in isolated cat papillary muscles stretched from 92 to 98% of the length at which maximal twitch force was developed (L(max)). The SFR was 123 +/- 1% of the immediate rapid phase (n = 6, P < 0.05) and was blunted by preventing EGFR transactivation with the Src-kinase inhibitor PP1 (99 +/- 2%, n = 4), matrix metalloproteinase inhibitor MMPI (108 +/- 4%, n = 11), the EGFR blocker AG1478 (98 +/- 2%, n = 6) or the mitochondrial transition pore blocker clyclosporine (99 +/- 3%, n = 6). Stretch increased ERK1/2 phosphorylation by 196 +/- 17% of control (n = 7, P < 0.05), an effect that was prevented by PP1 (124 +/- 22%, n = 7) and AG1478 (131 +/- 17%, n = 4). In myocardial slices, Ang II (which enhances ET mRNA) or endothelin-1 (ET-1)-induced increase in O(2)() production (146 +/- 14%, n = 9, and 191 +/- 17%, n = 13, of control, respectively, P < 0.05) was cancelled by AG1478 (94 +/- 5%, n = 12, and 98 +/- 15%, n = 8, respectively) or PP1 (100 +/- 4%, n = 6, and 99 +/- 8%, n = 3, respectively). EGF increased O(2)() production by 149 +/- 4% of control (n = 9, P < 0.05), an effect cancelled by inhibiting NADPH oxidase with apocynin (110 +/- 6% n = 7), mKATP channels with 5-hydroxydecanoic acid (5-HD; 105 +/- 5%, n = 8), the respiratory chain with rotenone (110 +/- 7%, n = 7) or the mitochondrial permeability transition pore with cyclosporine (111 +/- 10%, n = 6). EGF increased ERK1/2 phosphorylation (136 +/- 8% of control, n = 9, P < 0.05), which was blunted by 5-HD (97 +/- 5%, n = 4), suggesting that ERK1/2 activation is downstream of mitochondrial oxidative stress. Finally, stretch increased Ser703 Na(+)/H(+) exchanger-1 (NHE-1) phosphorylation by 172 +/- 24% of control (n = 4, P < 0.05), an effect that was cancelled by AG1478 (94 +/- 17%, n = 4). In conclusion, our data show for the first time that EGFR transactivation is crucial in the chain of events leading to the Anrep effect.
弗兰克-斯塔林机制 followed by 缓慢力反应 (SFR) 或安雷普效应。在这项研究中,我们假设 SFR 取决于心肌拉伸诱导的血管紧张素 II (Ang II)/内皮素 (ET) 释放后的表皮生长因子受体 (EGFR) 转激活。实验在从最大收缩力产生的长度的 92%拉伸到 98%的分离的猫乳头肌中进行。SFR 是立即快速相的 123 +/- 1% (n = 6, P < 0.05),并且通过使用Src-kinase 抑制剂 PP1 (99 +/- 2%, n = 4)、基质金属蛋白酶抑制剂 MMPI (108 +/- 4%, n = 11)、EGFR 阻滞剂 AG1478 (98 +/- 2%, n = 6) 或线粒体过渡孔阻滞剂环孢菌素 (99 +/- 3%, n = 6) 来阻止 EGFR 转激活来使 SFR 减弱。拉伸使 ERK1/2 磷酸化增加 196 +/- 17% of control (n = 7, P < 0.05),这种作用被 PP1 (124 +/- 22%, n = 7) 和 AG1478 (131 +/- 17%, n = 4) 阻止。在心肌切片中,Ang II(增强 ET mRNA)或内皮素-1 (ET-1) 诱导的 O(2)()产生增加 (146 +/- 14%, n = 9, 和 191 +/- 17%, n = 13, 分别为对照的,P < 0.05) 被 AG1478 (94 +/- 5%, n = 12, 和 98 +/- 15%, n = 8, 分别) 或 PP1 (100 +/- 4%, n = 6, 和 99 +/- 8%, n = 3, 分别) 取消。EGF 使 O(2)()产生增加 149 +/- 4% of control (n = 9, P < 0.05),这种作用被 NADPH 氧化酶抑制剂 apocynin (110 +/- 6% n = 7)、mKATP 通道抑制剂 5-羟基癸酸 (5-HD; 105 +/- 5%, n = 8)、呼吸链抑制剂鱼藤酮 (110 +/- 7%, n = 7) 或线粒体通透性过渡孔抑制剂环孢菌素 (111 +/- 10%, n = 6) 取消。EGF 使 ERK1/2 磷酸化增加 136 +/- 8% of control (n = 9, P < 0.05),这被 5-HD (97 +/- 5%, n = 4) 减弱,表明 ERK1/2 激活是线粒体氧化应激的下游。最后,拉伸使 Ser703 Na(+)/H(+) 交换器-1 (NHE-1) 磷酸化增加 172 +/- 24% of control (n = 4, P < 0.05),这种作用被 AG1478 (94 +/- 17%, n = 4) 取消。总之,我们的数据首次表明,EGFR 转激活在导致安雷普效应的事件链中是至关重要的。