P2Y receptor-mediated Ca2+ signaling increases human vascular endothelial cell permeability
P2Y receptor-mediated Ca2+ signaling increases human vascular endothelial cell permeability
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DOI：
10.1254/jphs.fpj03036x
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发表时间：
2004-06-01
影响因子：
3.5
通讯作者：
Shinozuka, K
中科院分区：
文献类型：
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作者：
Tanaka, N;Kawasaki, K;Shinozuka, K
我们研究了 P2 受体激动剂对细胞大小、细胞内钙水平 ([Ca2+](i)) 和 FITC 标记的葡聚糖 (FD-4) 渗透的影响，以及这些影响在人脐静脉内皮细胞 (HUVEC) 中之间的关系。 FD-4 浓度、细胞大小和 [Ca2+] 分别通过 HPLC 结合荧光、相差显微成像和荧光共焦显微成像进行分析。 P2Y(1) 受体激动剂 2-甲硫基 ATP (2meS-ATP) 和 ADP 可减小 HUVEC 中的细胞大小并增加 [Ca2+](i)。 P2Y(2)-受体激动剂UTP 增加[Ca2+](i)，但不影响细胞大小。 P2X受体激动剂α,β-亚甲基ATP不诱导这两种反应。 2meS-ATP 引起的尺寸减小和 [Ca2+](i) 增加可被磷酸吡哆醛-6-偶氮苯基-2',4'二磺酸（PPADS，P2Y(1)-拮抗剂）、毒胡萝卜素（Ca2+-泵抑制剂）和 U73122（磷脂酶 C 抑制剂）阻断。此外，2meS-ATP（P2Y(1)-受体激动剂）增强了 FD-4 通过内皮细胞单层的渗透。 PPADS、毒胡萝卜素和 U73122 也可以阻止 2meS-ATP 诱导的渗透增强。这些结果表明，P2Y 受体的激活会导致细胞大小减小、[Ca2+]i 增加，并可能参与促进 HUVEC 中的大分子通透性。
We investigated the effects of P2-receptor agonists on cell size, intracellular calcium levels ([Ca2+](i)), and permeation of FITC-labeled dextran (FD-4) as well as the relationship between these effects in human umbilical vein endothelial cells (HUVEC). FD-4 concentration, cell size, and [Ca2+], were analyzed by HPLC with fluorescence, phase contrast microscopic imaging, and fluorescent confocal microscopic imaging, respectively. The P2Y(1)-receptor agonists 2-methylthio ATP (2meS-ATP) and ADP decreased cell size and increased [Ca2+](i) in HUVEC. The P2Y(2)-receptor agonist UTP increased [Ca2+](i), but did not influence cell size. The P2X-receptor agonist alpha,beta-methylene ATP did not induce either response. The decrease in size and increase in [Ca2+](i) by 2meS-ATP were blocked by pyridoxalphosphate-6-azophenyl-2',4'disulphonic acid (PPADS, P2Y(1)-antagonist), thapsigargin (Ca2+-pump inhibitor), and U73122 (phospholipase C inhibitor). Furthermore, 2meS-ATP (P2Y(1)-receptor agonist) enhanced permeation of FD-4 through the endothelial cell monolayer. The 2meS-ATP-induced enhancement of the permeation was also prevented by PPADS, thapsigargin, and U73122. These results indicate that activation of P2Y receptors induces a decrease in cell size, an increase in [Ca2+] i, and may participate in facilitating macromolecular permeability in HUVEC.