Cardiac fibroblasts have functional TRPV4 activated by 4α-phorbol 12,13-didecanoate
Cardiac fibroblasts have functional TRPV4 activated by 4α-phorbol 12,13-didecanoate
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DOI：
10.1016/j.lfs.2009.10.013
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发表时间：
2009-12-16
期刊：
影响因子：
6.1
通讯作者：
Muraki, Katsuhiko
中科院分区：
文献类型：
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作者：
Hatano, Noriyuki;Itoh, Yuka;Muraki, Katsuhiko
香草酸型瞬时受体电位通道（TRPV）可能是多种细胞对多种刺激的潜在环境传感器。本研究首次证实了大鼠心脏成纤维细胞（CFs）中存在功能性香草酸4型瞬时受体电位通道（TRPV 4）。使用选择性TRPV 4激动剂4 α-佛波醇12，13-二癸酸酯（4 α PDD）评估TRPV 4在Us中的功能，同时测量细胞内Ca 2+浓度（[Ca 2 +]（i））和膜电流。CFs中也检测到TRPV 4蛋白的显著免疫反应性。当4 α PDD应用于CF时，[Ca 2 +]（i）以浓度依赖性方式升高。[Ca ~（2+）]（i）的升高被去除外部Ca ~（2+）和钌红（RuR）所消除。4 α PDD也激活非选择性阳离子电流（NSCC），这是由RuR抑制。此外，用靶向TRPV 4的短干扰RNA（siRNA）预处理Us显著降低了4 α PDD诱导的[Ca 2 +]（i）和NSCC的升高。(C)2009 Elsevier Inc. All rights reserved.
Aims: Vanilloid type transient receptor potential channel (TRPV) could be a potential environmental sensor to multiple stimuli in many types of cells. In this Study, we provide the first evidence of functional vanilloid type 4 transient receptor potential channel (TRPV4) in rat cardiac fibroblasts (CFs).Main methods: Expression of TRPV4 in Us was analyzed at mRNA and protein level. Function of TRPV4 in Us was evaluated using a selective TRPV4 agonist, 4 alpha-phorbol 12,13-didecanoate (4 alpha PDD) while measuring intracellular Ca2+ concentration ([Ca2+](i)) and membrane currents.Key findings: Analysis of expression of mRNA transcripts of TRPV subfamily revealed that TRPV2 and TRPV4 were expressed in Us. Significant immunoreactivity to TRPV4 protein was also detected in CFs. When 4 alpha PDD was applied to CFs, [Ca2+](i) was elevated in a concentration-dependent manner. The elevation of [Ca2+](i) was abolished by the removal of external Ca2+ and by ruthenium red (RuR). 4 alpha PDD also activated non-selective cation currents (NSCCs), which were suppressed by RuR. Moreover, pretreatment of Us with short interference RNA (siRNA) targeting TRPV4 significantly reduced both 4 alpha PDD-induced elevation of [Ca2+](i) and NSCC.Significance: These results provide strong evidence that endogenous TRPV4 functions as an important regulator of [Ca2+](i) in CFs. (C) 2009 Elsevier Inc. All rights reserved.