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首页> 外文期刊>American Journal of Physiology >RGD and YIGSR synthetic peptides facilitate cellular adhesion identical to that of laminin and fibronectin but alter the physiology of neonatal cardiac myocytes.
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RGD and YIGSR synthetic peptides facilitate cellular adhesion identical to that of laminin and fibronectin but alter the physiology of neonatal cardiac myocytes.

机译：RGD和YIGSR合成肽促进与层粘连蛋白和纤连蛋白相同的细胞粘附，但会改变新生儿心肌细胞的生理学。

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摘要

In the mammalian heart, the extracellular matrix plays an important role in regulating cell behavior and adaptation to mechanical stress. In cell culture, a significant number of cells detach in response to mechanical stimulation, limiting the scope of such studies. We describe a method to adhere the synthetic peptides RGD (fibronectin) and YIGSR (laminin) onto silicone for culturing primary cardiac cells and studying responses to mechanical stimulation. We first examined cardiac cells on stationary surfaces and observed the same degree of cellular adhesion to the synthetic peptides as their respective native proteins. However, the number of striated myocytes on the peptide surfaces was significantly reduced. Focal adhesion kinase (FAK) protein was reduced by 50% in cardiac cells cultured on YIGSR peptide compared with laminin, even though beta(1)-integrin was unchanged. Connexin43 phosphorylation increased in cells adhered to RGD and YIGSR peptides. We then subjected the cardiac cells to cyclic strainat 20% maximum strain (1 Hz) for 48 h. After this period, cell attachment on laminin was reduced to approximately 50% compared with the unstretched condition. However, in cells cultured on the synthetic peptides, there was no significant difference in cell adherence after stretch. On YIGSR peptide, myosin protein was decreased by 50% after mechanical stimulation. However, total myosin was unchanged in cells stretched on laminin. These results suggest that RGD and YIGSR peptides promote the same degree of cellular adhesion as their native proteins; however, they are unable to promote the signaling required for normal FAK expression and complete sarcomere formation in cardiac myocytes.
机译：在哺乳动物心脏中，细胞外基质在调节细胞行为和适应机械压力方面起着重要作用。在细胞培养中，大量细胞响应机械刺激而分离，从而限制了此类研究的范围。我们描述了一种方法，将合成肽RGD（纤连蛋白）和YIGSR（laminin）粘附到有机硅上，用于培养原代心脏细胞和研究对机械刺激的反应。我们首先检查了固定表面上的心肌细胞，并观察到细胞对合成肽的粘附程度与它们各自的天然蛋白质相同。然而，肽表面上的横纹肌细胞的数量显着减少。与层粘连蛋白相比，YIGSR肽培养的心肌细胞中的粘着斑激酶（FAK）蛋白减少了50％，即使beta（1）-整合素没有变化。连接到RGD和YIGSR肽的细胞中Connexin43磷酸化增加。然后，我们使心肌细胞经受最大应变20％（1 Hz）的循环应变48小时。在这段时间之后，层粘连蛋白上的细胞附着与未拉伸状态相比降低了约50％。但是，在合成肽上培养的细胞中，拉伸后细胞粘附没有显着差异。在YIGSR肽上，肌球蛋白在机械刺激后减少了50％。然而，层粘连蛋白拉伸的细胞中总肌球蛋白没有变化。这些结果表明RGD和YIGSR肽与它们的天然蛋白促进细胞粘附的程度相同。但是，它们不能促进正常FAK表达和心肌细胞中完整的肌节形成所需的信号传导。

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