Multiple Ca2+ Signaling Pathways Regulate Intracellular Ca2+ Activity in Human Cardiac Fibroblasts
Multiple Ca2+ Signaling Pathways Regulate Intracellular Ca2+ Activity in Human Cardiac Fibroblasts
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DOI：
10.1002/jcp.22010
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发表时间：
2010-04-01
影响因子：
5.6
通讯作者：
Li, Gui-Rong
中科院分区：
文献类型：
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作者：
Chen, Jing-Bo;Tao, Rong;Li, Gui-Rong
Ca 2+信号通路在心肌细胞中得到充分研究，但在心脏成纤维细胞中没有。本研究的目的是利用共聚焦扫描显微镜和RT-PCR技术来表征培养的人心脏成纤维细胞中的Ca 2+信号通路。发现约29%的人心脏成纤维细胞存在自发性胞内Ca ~（2+）振荡，而加入3%胎牛血清后，具有Ca ~（2+）振荡的细胞数增加到57.3%。Ca-1（2+）。振荡依赖于Ca 2+进入。钙池操纵的Ca ~（2+）内流通道阻断剂La ~（3+）、磷脂酶C抑制剂U-73122和三磷酸肌醇受体（IP_3Rs）抑制剂2-氨基乙氧基二苯基硼酸盐（2-aminoethoxydiphenylborate，IP_3Rs）都能抑制Ca ~（2+）振荡，但兰尼定不能。IP 3R激动剂硫柳汞可增强Ca ~（2+）振荡。抑制质膜Ca ~（2+）泵（PMCA）和Na ~+-Ca ~（2+）交换器（NCX）也能抑制Ca ~（2+）振荡。此外，硝苯地平可降低自发性钙振荡细胞的钙振荡频率，Bay K8644可增加自发性钙振荡细胞的钙振荡频率。RT-PCR显示，IP 3R 1 -3、SERCA 1 -3、Ca（v）1.2、NCX 3、PMCA 1，3，4、TRPC 1，3，4，6、STIM 1和Orai 1 -3的mRNA很容易检测到，但不能检测到RyR。我们的研究结果首次表明，自发的Ca ~（2+）振荡存在于培养的人心脏成纤维细胞中，并受到多种Ca ~（2+）途径的调节，这与充分研究的收缩性心肌细胞的Ca ~（2+）途径不同。本研究为进一步研究Ca 2+信号如何调节人心脏成纤维细胞的生物活性和病理条件下的心脏重构提供了基础。J.细胞。223：68-75，2010。(C)2009 Wiley-Liss，Inc.
Ca2+ signaling pathways are well studied in cardiac myocytes, but not in cardiac fibroblasts. The aim of the present study is to characterize Ca2+ signaling pathways in cultured human cardiac fibroblasts using confocal scanning microscope and RT-PCR techniques. It was found that spontaneous intracellular Ca2+ (Ca-i(2+)) oscillations were present in about 29% of human cardiac fibroblasts, and the number of cells with Ca-i(2+) oscillations was increased to 57.3% by application of 3% fetal bovine serum. Ca-i(2+). oscillations were dependent on Ca2+ entry. Ca-i(2+) oscillations were abolished by the store-operated Ca2+ (SOC) entry channel blocker La3+, the phospholipase C inhibitor U-73122, and the inositol trisphosphate receptors (IP3Rs) inhibitor 2-aminoethoxydiphenyl borate, but not by ryanodine. The IP3R agonist thimerosal enhanced Ca-i(2+) oscillations. Inhibition of plasma membrane Ca2+ pump (PMCA) and Na+-Ca2+ exchanger (NCX) also suppressed Ca-i(2+) oscillations. In addition, the frequency of Ca-i(2+) oscillations was reduced by nifedipine, and increased by Bay K8644 in cells with spontaneous Ca2+ oscillations. RT-PCR revealed that mRNAs for IP3R1-3, SERCA1-3, Ca(v)1.2, NCX3, PMCA1,3,4, TRPC1,3,4,6, STIM1, and Orai1-3, were readily detectable, but not RyRs. Our results demonstrate for the first time that spontaneous Ca-i(2+) oscillations are present in cultured human cardiac fibroblasts and are regulated by multiple Ca2+ pathways, which are not identical to those of the well-studied contractile cardiomyocytes. This study provides a base for future investigations into how Ca2+ signals regulate biological activity in human cardiac fibroblasts and cardiac remodeling under pathological conditions. J. Cell. Physiol. 223: 68-75, 2010. (C) 2009 Wiley-Liss, Inc.