Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors
Bone marrow-derived regenerated cardiomyocytes (CMG cells) express functional adrenergic and muscarinic receptors
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DOI：
10.1161/hc0302.102593
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发表时间：
2002-01-22
期刊：
影响因子：
37.8
通讯作者：
Ogawa, S
中科院分区：
文献类型：
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作者：
Hakuno, D;Fukuda, K;Ogawa, S
背景-我们最近报道了骨髓间充质干细胞经5-氮胞苷处理后可以在体外分化为心肌细胞。在天然心肌细胞中，肾上腺素能受体和毒蕈碱受体在介导心率、传导速度、收缩性和心脏肥大中起关键作用。我们研究了这些受体是否在分化的CMG细胞中表达，如果是，它们是否有下游信号系统，方法和结果-逆转录-聚合酶链反应显示，CMG细胞已经表达α（1A）-，α（1B）-和α（1D）-肾上腺素能受体mRNA 5-氮杂胞苷治疗前。而β（1）、β（2）-肾上腺素能受体和M-1-、M-2-毒蕈碱受体的表达在第1天首次检测到。苯肾上腺素剂量依赖性地诱导ERK 1/2磷酸化，哌唑嗪可完全抑制其磷酸化，异丙肾上腺素使cAMP增加38倍，普萘洛尔可完全抑制其磷酸化，异丙肾上腺素（10（-7）mol/L）使自发搏动频率增加47.6%（基础，127+/-16 bpm），普萘洛尔和CGP 20712 A（β（1）-选择性阻滞剂）分别使其降低79.0%和71.0%，而ICI 118551（β（2）-选择性阻滞剂）诱导轻微降低。细胞运动。对异丙肾上腺素的反应，收缩百分比和收缩速度分别增加37.5%，26.9%和50.6%。苯肾上腺素和异丙肾上腺素增加ANP和BNP基因表达。卡巴胆碱增加IP 3的32倍，这是显着抑制阿托品，以及AFDX 116（M-2-选择性阻滞剂）测量通过放射免疫测定。结论-这些研究结果表明，CMG细胞表达α（1A），α（1B）和α（1D）受体分化前和表达β（1），β（2），M-1和M-2受体后，他们获得心肌细胞表型。这些受体具有功能性信号传导途径，可以调节细胞功能。
Background-We recently reported that cardiomyocytes could be differentiated from bone marrow mesenchymal stem cells in vitro by 5-azacytidine treatment. In native cardiomyocytes, adrenergic and muscarinic receptors play crucial roles in mediating heart rate, conduction velocity, contractility, and cardiac hypertrophy. We investigated whether these receptors are expressed in differentiated CMG cells, and if so, whether they have downstream signaling systems,Methods and Results-Reverse transcription-polymerase chain reaction revealed that CMG cells had already expressed alpha(1A)-, alpha(1B)-, and alpha(1D)-adrenergic receptor mRNA before 5-azacytidine treatment. whereas expression of beta(1), beta(2)-adrenergic and M-1-, M-2-muscarinic receptors was first detected at 1 day. Phenylephrine dose-dependently induced phosphorylation of ERK1/2, which was completely inhibited by prazosin, and significantly increased cell size, Isoproterenol augmented cAMP by 38-fold, which was fully inhibited by propranolol, Isoproterenol (10(-7) mol/L) increased the spontaneous beating rate by 47.6% (basal, 127+/-16 bpm), and propranolol and CGP20712A (beta(1)-selective blocker) reduced it by 79.0% and 71.0%, respectively, whereas ICI118551 (beta(2)-selective blocker) induced slight reduction. Cell motion. percent shortening, and contractile velocity were increased by 37.5%, 26.9% and 50.6%, respectively, in response to isoproterenol. Phenylephrine and isoproterenol augmented ANP and BNP gene expressions. Carbachol increased IP3 by 32-fold, which was markedly inhibited by atropine as well as AFDX116 (M-2-selective blocker) measured by radioimmunoassay.Conclusions-These findings indicate that CMG cells expressed alpha(1A), alpha(1B), and alpha(1D) receptors before differentiation and expressed beta(1), beta(2), M-1, and M-2 receptors after they obtained the cardiomyocyte phenotype. These receptors had functional signal transduction pathways and could modulate cell function.