Ca2+-independent positive molecular inotropy for failing rabbit and human cardiac muscle by α-myosin motor gene transfer
Ca2+-independent positive molecular inotropy for failing rabbit and human cardiac muscle by α-myosin motor gene transfer
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DOI：
10.1096/fj.09-140566
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发表时间：
2010-02-01
期刊：
影响因子：
4.8
通讯作者：
Metzger, Joseph M.
中科院分区：
文献类型：
--
作者：
Herron, Todd J.;Devaney, Eric;Metzger, Joseph M.
目前的正性肌力疗法用于通过增加可用于收缩的钙量来增强衰竭心脏中心的心肌收缩力，但长期使用它们会因致命性心律失常而导致死亡率增加。因此，需要开发和探索可以通过钙非依赖性机制发挥作用的新型正性肌力疗法。本研究的目的是确定快速α-肌球蛋白分子运动基因转移是否可以在慢速β-肌球蛋白主导的兔子和人类衰竭心室肌细胞中赋予钙独立的正性肌力。为此，我们生成了重组腺病毒（AdMYH6），可在体外将全长人α-肌球蛋白基因传递至成年兔和人心肌细胞。通过蛋白质印迹、免疫细胞化学分析和共聚焦成像测定快速α-肌球蛋白运动表达。在使用来自缺血性衰竭心脏的电刺激肌细胞的实验中，AdMYH6 增加了衰竭人类 [23.9 +/- 7.8 nm (n = 10) vs. AdMYH6 振幅 78.4 +/- 16.5 nm (n = 6)] 和兔肌细胞的收缩幅度。细胞内钙瞬变幅度没有改变。对照实验包括使用绿色荧光蛋白或β-肌球蛋白重链腺病毒。我们的数据为通过快速α-肌球蛋白运动蛋白基因转移在衰竭心肌细胞中形成一种新型的不依赖于钙的正性肌力提供了证据。-Herron, T. J., Devaney, E., Mundada, L., Arden, E., Day, S., Guerrero-Serna, G., Turner, I., Westfall, M., Metzger, J. M. Ca2+独立的正性分子正性肌力对于衰竭心肌细胞的作用通过α-肌球蛋白运动基因转移兔和人心肌。 FASEB J. 24, 415-424 (2010)。 www.fasebj.org
Current inotropic therapies used to increase cardiac contractility of the failing heart center on increasing the amount of calcium available for contraction, but their long-term use is associated with increased mortality due to fatal arrhythmias. Thus, there is a need to develop and explore novel inotropic therapies that can act via calcium-independent mechanisms. The purpose of this study was to determine whether fast alpha-myosin molecular motor gene transfer can confer calcium-independent positive inotropy in slow beta-myosin-dominant rabbit and human failing ventricular myocytes. To this end, we generated a recombinant adenovirus (AdMYH6) to deliver the full-length human alpha-myosin gene to adult rabbit and human cardiac myocytes in vitro. Fast alpha-myosin motor expression was determined by Western blotting and immunocytochemical analysis and confocal imaging. In experiments using electrically stimulated myocytes from ischemic failing hearts, AdMYH6 increased the contractile amplitude of failing human [23.9 +/- 7.8 nm (n = 10) vs. AdMYH6 amplitude 78.4 +/- 16.5 nm (n = 6)] and rabbit myocytes. The intracellular calcium transient amplitude was not altered. Control experiments included the use of a green fluorescent protein or a beta-myosin heavy chain adenovirus. Our data provide evidence for a novel form of calcium-independent positive inotropy in failing cardiac myocytes by fast alpha-myosin motor protein gene transfer.-Herron, T. J., Devaney, E., Mundada, L., Arden, E., Day, S., Guerrero-Serna, G., Turner, I., Westfall, M., Metzger, J. M. Ca2+-independent positive molecular inotropy for failing rabbit and human cardiac muscle by alpha-myosin motor gene transfer. FASEB J. 24, 415-424 (2010). www.fasebj.org