Role of myosin light chain kinase in cardiotrophin-1-induced cardiac myofibroblast cell migration
Role of myosin light chain kinase in cardiotrophin-1-induced cardiac myofibroblast cell migration
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DOI：
10.1152/ajpheart.01041.2010
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发表时间：
2011-08-01
影响因子：
4.8
通讯作者：
Dixon, Ian M. C.
中科院分区：
文献类型：
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作者：
Freed, Darren H.;Chilton, Lisa;Dixon, Ian M. C.
Freed DH、Chilton L、Li Y、Dangerfield AL、Raizman JE、Rattan SG、Visen N、Hryshko LV、Dixon IM。肌球蛋白轻链激酶在心肌营养蛋白-1诱导的心肌成纤维细胞迁移中的作用。 Am J Physiol Heart Circ Physiol 301：H514-H522，2011。首次发表于 2011 年 5 月 13 日； doi:10.1152/ajpheart.01041.2010.-肌成纤维细胞的趋化运动被认为是其隔离到组织损伤部位的常用手段。心肌梗塞（MI）后，将心肌成纤维细胞募集到梗塞疤痕是伤口愈合的关键步骤。收缩性肌成纤维细胞表达胚胎平滑肌肌球蛋白、α-平滑肌肌动蛋白以及 I 型和 III 型胶原蛋白。我们检查了心肌营养素-1 (CT-1) 在诱导原代大鼠心室肌成纤维细胞运动中的作用。研究膜电位（E（m））和Ca（2+）进入的变化以揭示诱导肌成纤维细胞迁移的机制。 CT-1 诱导的心肌成纤维细胞迁移，通过抑制 JAK2 (25 mu M AG490) 和肌球蛋白轻链激酶 (20 mu M ML-7) 减弱。 CT-1 存在时，10 μM 钆 (Gd(3+)) 对 K(+) 通道（1 mM 四乙铵或 100 μM 4-氨基吡啶）和非选择性阳离子通道的抑制显着减少迁移。 CT-1处理导致肌球蛋白轻链磷酸化显着增加，可通过在无Ca(2+)条件下孵育或应用AG490、ML-7和W7（100μM；钙调蛋白抑制剂）来抑制这种磷酸化。用电位荧光 DiBAC(4)(3) 染料监测肌成纤维细胞膜电位，揭示了对 CT-1 的双相反应，包括初始去极化和随后的超极化。通过荧光 3 评估，细胞内 Ca(2+) 增加，在膜去极化后立即发生，并在最大超极化时减弱。 CT-1 通过心室肌成纤维细胞中的多种并行信号传导方式发挥趋化作用，包括膜电位的变化、细胞内钙的改变以及许多细胞内信号传导途径的激活。需要进一步研究以确定 K(+) 电流在此过程中的精确作用。
Freed DH, Chilton L, Li Y, Dangerfield AL, Raizman JE, Rattan SG, Visen N, Hryshko LV, Dixon IM. Role of myosin light chain kinase in cardiotrophin-1-induced cardiac myofibroblast cell migration. Am J Physiol Heart Circ Physiol 301: H514-H522, 2011. First published May 13, 2011; doi:10.1152/ajpheart.01041.2010.-Chemotactic movement of myofibroblasts is recognized as a common means for their sequestration to the site of tissue injury. Following myocardial infarction (MI), recruitment of cardiac myofibroblasts to the infarct scar is a critical step in wound healing. Contractile myofibroblasts express embryonic smooth muscle myosin, alpha-smooth muscle actin, as well as collagens I and III. We examined the effects of cardiotrophin-1 (CT-1) in the induction of primary rat ventricular myofibroblast motility. Changes in membrane potential (E(m)) and Ca(2+) entry were studied to reveal the mechanisms for induction of myofibroblast migration. CT-1-induced cardiac myofibroblast cell migration, which was attenuated through the inhibition of JAK2 (25 mu M AG490), and myosin light chain kinase (20 mu M ML-7). Inhibition of K(+) channels (1 mM tetraethylammonium or 100 mu M 4-aminopyridine) and nonselective cation channels by 10 mu M gadolinium (Gd(3+)) significantly reduced migration in the presence of CT-1. CT-1 treatment caused a significant increase in myosin light chain phosphorylation, which could be inhibited by incubation in Ca(2+)-free conditions or by application of AG490, ML-7, and W7 (100 mu M; calmodulin inhibitor). Monitoring myofibroblast membrane potential with potentiometric fluorescent DiBAC(4)(3) dye revealed a biphasic response to CT-1 consisting of an initial depolarization followed by hyperpolarization. Increased intracellular Ca(2+), as assessed by fluo 3, occurred immediately after membrane depolarization and attenuated at the time of maximal hyperpolarization. CT-1 exerts chemotactic effects via multiple parallel signaling modalities in ventricular myofibroblasts, including changes in membrane potential, alterations in intracellular calcium, and activation of a number of intracellular signaling pathways. Further study is warranted to determine the precise role of K(+) currents in this process.