P2Y1 and P2Y13 purinergic receptors mediate Ca2+ signaling and proliferative responses in pulmonary artery vasa vasorum endothelial cells
P2Y1 and P2Y13 purinergic receptors mediate Ca2+ signaling and proliferative responses in pulmonary artery vasa vasorum endothelial cells
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DOI：
10.1152/ajpcell.00237.2010
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发表时间：
2011-02-01
影响因子：
5.5
通讯作者：
Gerasimovskaya, Evgenia V.
中科院分区：
文献类型：
--
作者：
Lyubchenko, Taras;Woodward, Heather;Gerasimovskaya, Evgenia V.
Lyubchenko T，Woodward H，Veo KD，Burns N，Nijmeh H，Liubchenko GA，Stenmark KR，Gerasimovskaya EV. P2 Y1和P2 Y13嘌呤能受体介导肺动脉血管内皮细胞的Ca ~（2+）信号和增殖反应。Am J Physiol Cell Physiol 300：C266-C275，2011.首次发表于2010年10月20日; doi：10.1152/ajpcell.00237.2010.-细胞外ATP和ADP对肺动脉血管外膜血管内皮细胞（VVEC）具有强的促血管生成作用。然而，细胞外核苷酸介导的血管生成的分子信号转导机制仍然没有完全阐明。由于细胞内Ca 2+浓度（[Ca 2 +]（i））的升高是细胞增殖所必需的，并且响应于细胞外核苷酸而发生，因此进行本研究以描述参与VVEC中Ca 2+信号传导和细胞外核苷酸介导的促有丝分裂反应的嘌呤能受体亚型。我们的数据表明，用胞外ATP刺激VVEC导致[Ca 2 +]（i）通过质膜通道的Ca 2+内流以及胞内储存的Ca 2+动员而升高。此外，细胞外ATP诱导同时在胞质和核室的Ca 2+的反应。观察到[Ca 2 +]（i）对多种嘌呤能受体激动剂的反应增加，包括ATP、ADP、ATP γ S、ADP β S、UTP、UDP、2-甲硫基-ATP（MeSATP）、2-甲硫基-ADP（MeSADP）和BzATP，但不包括腺苷、AMP、二腺苷四磷酸、α β MeATP和β γ MeATP。采用RT-PCR技术，我们鉴定了VVEC中P2 Y1、P2 Y2、P2 Y 4、P2 Y13、P2 Y14、P2 X2、P2 X5、P2 X7、A1、A2 b和A3嘌呤能受体的mRNA。与P2 Y1选择性拮抗剂MRS 2179和P2 Y13选择性拮抗剂MRS 2211以及百日咳毒素预孵育VVEC，在不同程度上减弱激动剂诱导的细胞内Ca 2+反应和ERK 1/2，Akt和S6核糖体蛋白的激活，表明P2 Y1和P2 Y13受体在VVEC生长反应中起主要作用。考虑到嘌呤能信号在调节血管生成和血管稳态中的广泛生理意义，我们的研究结果表明，P2 Y1和P2 Y13受体可能是治疗涉及血管扩张的病理性血管重塑的新的和特异性靶点。
Lyubchenko T, Woodward H, Veo KD, Burns N, Nijmeh H, Liubchenko GA, Stenmark KR, Gerasimovskaya EV. P2Y1 and P2Y13 purinergic receptors mediate Ca2+ signaling and proliferative responses in pulmonary artery vasa vasorum endothelial cells. Am J Physiol Cell Physiol 300: C266-C275, 2011. First published October 20, 2010; doi: 10.1152/ajpcell.00237.2010.-Extracellular ATP and ADP have been shown to exhibit potent angiogenic effects on pulmonary artery adventitial vasa vasorum endothelial cells (VVEC). However, the molecular signaling mechanisms of extracellular nucleotide-mediated angiogenesis remain not fully elucidated. Since elevation of intracellular Ca2+ concentration ([Ca2+](i)) is required for cell proliferation and occurs in response to extracellular nucleotides, this study was undertaken to delineate the purinergic receptor subtypes involved in Ca2+ signaling and extracellular nucleotide-mediated mitogenic responses in VVEC. Our data indicate that stimulation of VVEC with extracellular ATP resulted in the elevation of [Ca2+](i) via Ca2+ influx through plasma membrane channels as well as Ca2+ mobilization from intracellular stores. Moreover, extracellular ATP induced simultaneous Ca2+ responses in both cytosolic and nuclear compartments. An increase in [Ca2+](i) was observed in response to a wide range of purinergic receptor agonists, including ATP, ADP, ATP gamma S, ADP beta S, UTP, UDP, 2-methylthio-ATP (MeSATP), 2-methylthio-ADP (MeSADP), and BzATP, but not adenosine, AMP, diadenosine tetraphosphate, alpha beta MeATP, and beta gamma MeATP. Using RT-PCR, we identified mRNA for the P2Y1, P2Y2, P2Y4, P2Y13, P2Y14, P2X2, P2X5, P2X7, A1, A2b, and A3 purinergic receptors in VVEC. Preincubation of VVEC with the P2Y1 selective antagonist MRS2179 and the P2Y13 selective antagonist MRS2211, as well as with pertussis toxin, attenuated at varying degrees agonist-induced intracellular Ca2+ responses and activation of ERK1/2, Akt, and S6 ribosomal protein, indicating that P2Y1 and P2Y13 receptors play a major role in VVEC growth responses. Considering the broad physiological implications of purinergic signaling in the regulation of angiogenesis and vascular homeostasis, our findings suggest that P2Y1 and P2Y13 receptors may represent novel and specific targets for treatment of pathological vascular remodeling involving vasa vasorum expansion.