Abstract
Cardiomyocytes possess the ability to respond to mechanical stimuli by reprogramming their gene expression. This study investigated the effects of different loading protocols on signaling and expression responses of myogenic, anabolic, inflammatory, atrophy and pro-apoptotic genes in cardiomyocyte-like H9C2 cells. Differentiated H9C2 cells underwent various stretching protocols by altering their elongation, frequency and duration, utilizing an in vitro cell tension system. The loading-induced expression changes of MyoD, Myogenin, MRF4, IGF-1 isoforms, Atrogin-1, Foxo1, Fuca and IL-6 were measured by Real Time-PCR. The stretching-induced activation of Akt and Erk 1/2 was also evaluated by Western blot analysis. Low strain (2.7% elongation), low frequency (0.25 Hz) and intermediate duration (12 h) stretching protocol was overall the most effective in inducing beneficial responses, i.e., protein synthesis along with the suppression of apoptosis, inflammation and atrophy, in the differentiated cardiomyocytes. These findings demonstrated that varying the characteristics of mechanical loading applied on H9C2 cells in vitro can regulate their anabolic/survival program.
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We are grateful to Dr. Panagiotis Lembessis, National and Kapodistrian University of Athens, for the critical reading of the manuscript.
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The Authors declare that no external or extramural support was provided and funding was exclusively through the Department of Physiology, Medical School, National and Kapodistrian University of Athens.
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EZ designed and performed the experiments, analyzed the data and wrote the manuscript; AP and MK conceived or designed the experiments, analyzed data and wrote the manuscript; AM performed the experiments and analyzed the data; AC analyzed data and reviewed the manuscript. All Authors have read and approved the final manuscript.
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Zevolis, E., Philippou, A., Moustogiannis, A. et al. Optimizing mechanical stretching protocols for hypertrophic and anti-apoptotic responses in cardiomyocyte-like H9C2 cells. Mol Biol Rep 48, 645–655 (2021). https://doi.org/10.1007/s11033-020-06112-z
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DOI: https://doi.org/10.1007/s11033-020-06112-z