The effects of extracellular nucleotides on [Ca2+]i signalling in a human-derived renal proximal tubular cell line (HKC-8).
The effects of extracellular nucleotides on [Ca2+]i signalling in a human-derived renal proximal tubular cell line (HKC-8).
复制标题
细胞外核苷酸对人源肾近端小管细胞系 (HKC-8) 中 [Ca2]i 信号传导的影响。
DOI：
10.1002/jcb.22390
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发表时间：
2010
影响因子：
4
通讯作者：
Turvey MR
中科院分区：
文献类型：
--
作者：
Turvey MR
HKC-8细胞是人源性肾近端小管细胞系，为研究人肾细胞功能提供了有用的模型系统。在这项研究中，我们的目的是确定在HKC-8中由P2受体介导的[Ca 2 +] i信号传导。Fura-2和比率成像法测定HKC-8细胞内[Ca 2 +] i。我们的研究结果表明，ATP激活P2 Y受体可诱导依赖于细胞内Ca 2+来源的[Ca 2 +] i升高，而P2 Y受体的长期激活可诱导依赖于细胞内和细胞外Ca 2+来源的[Ca 2 +] i升高。本研究中的药理学和分子数据表明，TRPC 4通道介导HKC-8细胞中Ca 2+内流与P2 Y激活偶联。U 73221，一种PI-PLC的抑制剂，不抑制初始ATP诱导的反应;而D 609，一种PC-PLC的抑制剂，导致初始ATP诱导的反应显著降低，表明P2 Y受体与PC-PLC偶联。虽然P2 X在HKC-8中存在，但P2 X激动剂α，β me-ATP未能引起[Ca 2 +]i升高。然而，浓度为100 µM的PPADS抑制ATP诱导的[Ca 2 +]i升高。我们的结果表明，在HKC-8细胞中存在功能性P2 Y受体。ATP通过P2 Y诱导的[Ca 2 +] i升高与PC‐PLC和TRP通道密切相关。J.细胞。109：132-139，2010.© 2009 Wiley利斯公司
HKC‐8 cells are a human‐derived renal proximal tubular cell line and provide a useful model system for the study of human renal cell function. In this study, we aimed to determine [Ca2+]isignalling mediated by P2 receptor in HKC‐8. Fura‐2 and a ratio imaging method were employed to measure [Ca2+]iin HKC‐8 cells. Our results showed that activation of P2Y receptors by ATP induced a rise in [Ca2+]ithat was dependent on an intracellular source of Ca2+, while prolonged activation of P2Y receptors induced a rise in [Ca2+]ithat was dependent on intra‐ and extracellular sources of Ca2+. Pharmacological and molecular data in this study suggests that TRPC4 channels mediate Ca2+entry in coupling to activation of P2Y in HKC‐8 cells. U73221, an inhibitor of PI‐PLC, did not inhibit the initial ATP‐induced response; whereas D609, an inhibitor of PC‐PLC, caused a significant decrease in the initial ATP‐induced response, suggesting that P2Y receptors are coupled to PC‐PLC. Although P2X were present in HKC‐8, The P2X agonist, α,β me‐ATP, failed to cause a rise in [Ca2+]i. However, PPADS at a concentration of 100 µM inhibits the ATP‐induced rise in [Ca2+]i. Our results indicate the presence of functional P2Y receptors in HKC‐8 cells. ATP‐induced [Ca2+]ielevation via P2Y is tightly associated with PC‐PLC and TRP channel. J. Cell. Biochem. 109: 132–139, 2010. © 2009 Wiley‐Liss, Inc.