α1-adrenoceptor stimulation enhances leukemia inhibitory factor-induced proliferation. of mouse-induced pluripotent stem cells
α1-adrenoceptor stimulation enhances leukemia inhibitory factor-induced proliferation. of mouse-induced pluripotent stem cells
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α1-肾上腺素受体刺激增强白血病抑制因子诱导的小鼠诱导多能干细胞的增殖。
DOI：
10.1016/j.ejphar.2011.06.026
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发表时间：
2011
影响因子：
5
通讯作者：
Y.Watanabe
中科院分区：
文献类型：
--
作者：
T.Ishizuka;Y.Watanabe
由于诱导多能干细胞（iPS）的临床应用可能会克服目前的障碍，在干细胞为基础的治疗，调节iPS细胞增殖的分子机制是极大的兴趣。因此，在本研究中，我们确定了α1-肾上腺素受体参与小鼠iPS细胞的增殖。选择性α1-肾上腺素受体激动剂l-苯肾上腺素剂量依赖性地增加在白血病抑制因子（LIF）培养基中培养的小鼠iPS细胞的增殖。用选择性α1肾上腺素受体拮抗剂或蛋白激酶C（PKC）抑制剂预处理可显着抑制l-去氧肾上腺素诱导的DNA合成。用IP 3受体激动剂处理显著增强LIF诱导的DNA合成。另一方面，我们证实，细胞内钙水平增加的处理与l-苯乙醯胺。因此，α1-肾上腺素受体激活引起的细胞内钙释放或PKC激活可能导致DNA合成的增强。此外，用丝裂原活化蛋白激酶（MAPK）激酶（MEK）抑制剂PD 98059或磷脂酰肌醇-3磷酸激酶（PI 3 K）抑制剂LY 294002预处理显著抑制l-苯肾上腺素诱导的DNA合成。用l-苯丙氨酸处理显著增加Akt或p44/42 MAPK磷酸化。免疫荧光染色和Western blotting分析证实小鼠iPS细胞表达α1肾上腺素受体。在用LIF培养的小鼠iPS细胞中，用l-苯肾上腺素刺激显著增加S期和G2/M期细胞的比例，而减少G1期细胞的比例。这些结果表明，α1肾上腺素能受体的刺激可能通过增强MEK/MAPK和PI 3 K/Akt途径促进LIF诱导的iPS细胞DNA合成和增殖。
Since the clinical use of induced pluripotent stem (iPS) cells may overcome the current obstacles in stem cell-based therapy, the molecular mechanisms that regulate iPS cell proliferation are of great interest. Therefore, in the present study, we determined the involvement of α1-adrenoceptor in the proliferation of mouse iPS cells. The selective α1-adrenoceptor agonist l-phenylephrine dose-dependently increased the proliferation of mouse iPS cells cultured in a medium with leukemia inhibitory factor (LIF). Pretreatment with either selective α1-adrenoceptor antagonists or protein kinase C (PKC) inhibitors significantly inhibited l-phenylephrine-induced DNA synthesis. The treatment with an IP3receptor agonist significantly enhanced LIF-induced DNA synthesis. On the other hand, we confirmed that the intracellular calcium level was increased by the treatment with l-phenylephrine. Thus, intracellular calcium release or PKC activation induced by α1-adrenoceptor activation may lead to the enhancement of DNA synthesis. In addition, pretreatment with mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor PD98059 or phosphatidylinositol-3 phosphate kinase (PI3K) inhibitor LY294002 significantly inhibited l-phenylephrine-induced DNA synthesis. Treatment with l-phenylephrine significantly increased Akt or p44/42 MAPK phosphorylation. α1-Adrenoceptor expression in mouse iPS cells was confirmed by immunofluorescence staining and western blotting analysis. In mouse iPS cells cultured with LIF, stimulation with l-phenylephrine significantly increased the proportion of cells in the S and G2/M phases and decreased that in the G1phase. These results suggest that stimulation with α1-adrenoceptor may enhance DNA synthesis and proliferation of mouse iPS cells cultured with LIF via augmentation of both the MEK/MAPK and the PI3K/Akt pathways.