Department of Pharmacology, National Defense Medical College, 3-2, Namiki, Tokorozawa, Saitama 359-8513, Japan.
Department of Pharmacology, National Defense Medical College, 3-2, Namiki, Tokorozawa, Saitama 359-8513, Japan.
Eur J Pharmacol. 2014 Oct 5;740:28-34. doi: 10.1016/j.ejphar.2014.06.056. Epub 2014 Jul 9.
Previous studies suggest that β-adrenoceptor stimulation may enhance the cardiac differentiation of mouse embryonic stem (ES) cells. It remains unclear whether the differentiations of ES cells and induced pluripotent stem (iPS) cells rely on similar molecular mechanisms. In addition, no previous studies have shown that human iPS cells express β-adrenoceptors. Therefore, in the present study, we determined the involvement of β-adrenoceptors in the differentiation of human iPS cells into mesodermal progenitor cells. The induction of differentiation of human iPS cells into kinase insert domain receptor (KDR)-positive mesodermal progenitor cells was performed on feeder cells in a differentiation medium with basic fibroblast growth factor (bFGF), bone morphogenetic protein-4 (BMP-4), and activin A. When the iPS cells that were exposed to bFGF, BMP-4, and activin A were treated with L-isoproterenol (a β-adrenoceptor agonist) for 4 days, the expression of KDR was significantly increased compared to that in the cells that were not treated with L-isoproterenol. Pretreatment of the cells with either atenolol (a β1-adrenoceptor antagonist) or ICI-118551 (a β2-adrenoceptor antagonist) significantly inhibited the L-isoproterenol-induced increase in KDR expression. In addition, pretreatment with both H89 (a protein kinase A inhibitor) and SB203580 (a p38 MAPK inhibitor) significantly inhibited the L-isoproterenol-induced increase in KDR expression. Treatment with L-isoproterenol enhanced the phosphorylation of p38 MAPK in human iPS cells exposed to bFGF, BMP-4, and activin A. These results suggest that β-adrenoceptor stimulation in human iPS cells may enhance their differentiation into mesodermal progenitor cells via the activation of either protein kinase A or p38 MAPK.
先前的研究表明，β-肾上腺素受体的刺激可能会增强小鼠胚胎干细胞（ES 细胞）的心脏分化。目前尚不清楚 ES 细胞和诱导多能干细胞（iPS 细胞）的分化是否依赖于相似的分子机制。此外，以前的研究尚未表明人 iPS 细胞表达β-肾上腺素受体。因此，在本研究中，我们确定了β-肾上腺素受体是否参与人 iPS 细胞向中胚层祖细胞的分化。在含有碱性成纤维细胞生长因子（bFGF）、骨形态发生蛋白-4（BMP-4）和激活素 A 的分化培养基中，在饲养细胞上诱导人 iPS 细胞分化为激酶插入结构域受体（KDR）阳性中胚层祖细胞。当将暴露于 bFGF、BMP-4 和激活素 A 的 iPS 细胞用 L-异丙肾上腺素（β-肾上腺素受体激动剂）处理 4 天时，与未用 L-异丙肾上腺素处理的细胞相比，KDR 的表达显著增加。用阿替洛尔（β1-肾上腺素受体拮抗剂）或 ICI-118551（β2-肾上腺素受体拮抗剂）预处理细胞可显著抑制 L-异丙肾上腺素诱导的 KDR 表达增加。此外，用 H89（蛋白激酶 A 抑制剂）和 SB203580（p38 MAPK 抑制剂）预处理均可显著抑制 L-异丙肾上腺素诱导的 KDR 表达增加。用 L-异丙肾上腺素处理可增强暴露于 bFGF、BMP-4 和激活素 A 的人 iPS 细胞中 p38 MAPK 的磷酸化。这些结果表明，β-肾上腺素受体刺激可能通过激活蛋白激酶 A 或 p38 MAPK 增强人 iPS 细胞向中胚层祖细胞的分化。
