Effects of docosahexaenoic acid or arachidonic acid supplementation on gene expression and contractile force of rat cardiomyocytes in primary culture
Effects of docosahexaenoic acid or arachidonic acid supplementation on gene expression and contractile force of rat cardiomyocytes in primary culture
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DOI：
10.32604/biocell.2021.016281
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发表时间：
2021
期刊：
影响因子：
1.2
通讯作者：
Mizuna Yano;Yuta Umehara;Tomokazu Kudo;Takao Nakamura;T. Kosawada;Atsuyoshi Nishina;M. Sazuka;D. Sato;Zhonggang Feng
中科院分区：
文献类型：
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作者：
Mizuna Yano;Yuta Umehara;Tomokazu Kudo;Takao Nakamura;T. Kosawada;Atsuyoshi Nishina;M. Sazuka;D. Sato;Zhonggang Feng
虽然脂肪酸在心肌生理学中发挥重要作用，但传统培养基几乎不含脂质。我们之前发现，大鼠新生心肌主要含有二十二碳六烯酸（DHA）、亚油酸（LA）和花生四烯酸（AA）作为多不饱和脂肪酸（PUFA），并且来自胎鼠的培养心肌细胞中这些含量明显低于新生心肌中的含量。在本研究中，我们首先评估了补充 DHA、LA 或 AA 对原代培养的大鼠心肌细胞脂肪酸含量和收缩面积百分比变化的影响。在此评估的基础上，我们评估了 DHA 或 AA 补充剂对 mRNA 表达的影响，并进一步直接测量了补充剂后心肌细胞的收缩力。这项研究表明，在补充 20 μM DHA 或 50 μM AA 时，收缩面积的百分比变化最大，而补充 LA 并不影响该收缩指数，并且在补充 DHA 或 AA 的培养心肌细胞中，与分化、成熟度、脂肪酸代谢和细胞粘附相关的 mRNA 表达存在广泛的上调趋势。特别是，细胞粘附分子 connexin43 和 N-cadherin 基因表达的上调非常显着，而对分化和成熟的影响则不太明显。相应地，添加补充剂后培养皿中心肌细胞簇收缩面积的百分比变化显着增加，而收缩力的增强则不大。这些结果表明，向胎儿心肌细胞培养物中添加DHA或AA可有效防止培养物中心肌细胞的去分化，并且收缩性能的增强可能主要归因于细胞间连接的改善。
While fatty acids play essential roles in the physiology of the myocardium, conventional culture media contain little lipid. We previously revealed that rat neonatal myocardium mainly contains docosahexaenoic (DHA), linoleic (LA), and arachidonic (AA) acids as polyunsaturated fatty acids (PUFAs), and these contents in cultured cardiomyocytes derived from fetal rats were markedly lower than those in the neonatal myocardium. In this study, we first assessed the effects of supplementation of DHA, LA, or AA on the fatty acid contents and the percentage change of contractile area in primarily cultured rat cardiomyocytes. Based on this assessment, we then evaluated the effects of DHA or AA supplementation on mRNA expression and further directly measured the contractile force of cardiomyocytes with the supplementations. This study revealed that percentage change of contractile area was maximized under 20 μM DHA or 50 μM AA supplementation while LA supplementation did not affect this contraction index, and that a widespread upregulation tendency of the mRNA expression related to differentiation, maturity, fatty acid metabolism, and cell adhesion was seen in the cultured cardiomyocytes with supplementation of DHA or AA. In particular, upregulation of the gene expression of cellular adhesion molecules connexin43 and N-cadherin were remarkable, whereas the effects on differentiation and maturation were less pronounced. Correspondingly, the increase of the percentage change of the contractile area of cardiomyocyte clusters in culture dishes with the supplementations was significant, whereas the enhancement of the contractile force was modest. These results suggest that supplementation of DHA or AA to the fetal cardiomyocyte culture may play effective roles in preventing the de-differentiation of the cardiomyocytes in culture and that the enhancement of the contractile performance may be mainly attributed to the improvement of intercellular connection.