Adenosine-5′-triphosphate up-regulates proliferation of human cardiac fibroblasts
Adenosine-5′-triphosphate up-regulates proliferation of human cardiac fibroblasts
复制标题
DOI：
10.1111/j.1476-5381.2012.01831.x
复制
发表时间：
2012-06-01
影响因子：
7.3
通讯作者：
Li, Gui-Rong
中科院分区：
文献类型：
--
作者：
Chen, Jing-Bo;Liu, Wen-Juan;Li, Gui-Rong
背景和目的ATP是一种有效的信号分子，其调节生物活性，包括增加或减少不同类型细胞的增殖。本研究的目的是探讨ATP如何调节人心脏成纤维细胞的增殖。实验方法采用逆转录-聚合酶链反应（RT-PCR）、蛋白质印迹分析、细胞增殖和迁移实验研究ATP对人心室成纤维细胞的作用。关键结果ATP以浓度依赖性方式增加细胞增殖。同样，P2 X受体激动剂α，β-亚甲基ATP和P2 Y受体激动剂ATP-？S还上调细胞增殖。P2受体拮抗剂苏拉明和活性蓝-2阻止ATP诱导的增殖增加，RT-PCR和Western blot分析显示，P2 X4/7和P2 Y2的mRNA在心脏成纤维细胞中是丰富的。ATP增加磷酸化PKB（Akt）和ERK 1/2水平;苏拉明、活性蓝-2、PI 3-激酶抑制剂渥曼青霉素、PKB抑制剂API-2和MAPK抑制剂PD 98059可拮抗这种作用。这些激酶抑制剂也阻止了ATP诱导的增殖增加。此外，ATP通过增加细胞周期蛋白D1和细胞周期蛋白E的蛋白表达，促进细胞从G 0/G1期向S期的进展。用靶向相应受体的siRNA沉默P2 X4/7和P2 Y2受体减少了ATP刺激的心脏成纤维细胞增殖和迁移。结论ATP通过激活P2 X4/7和P2 Y2受体，促进细胞周期进程，从而上调人心脏成纤维细胞的增殖。它也增加了这些细胞的迁移。ATP的这些作用可能参与损伤心脏的心脏重构。
BACKGROUND AND PURPOSE ATP is a potent signalling molecule that regulates biological activities including increasing or decreasing proliferation in different types of cells. The aim of the present study was to investigate how ATP regulates the proliferation of human cardiac fibroblasts. EXPERIMENTAL APPROACH Reverse transcription (RT)-PCR, Western blot analysis, cell proliferation and migration assays were employed to investigate the effects of ATP on human adult ventricular fibroblasts. KEY RESULTS ATP increased cell proliferation in a concentration-dependent manner. Similarly, the P2X receptor agonist a,beta-methylene ATP and P2Y receptor agonist ATP-?S also up-regulated cell proliferation. The P2 receptor antagonists suramin and reactive blue-2 prevented the ATP-induced increase in proliferation and RT-PCR and Western blot analysis revealed that mRNAs of P2X4/7 and P2Y2 are abundant in cardiac fibroblasts. ATP increased phosphorylated PKB (Akt) and ERK1/2 levels; an effect antagonized by suramin, reactive blue-2, the PI3-kinase inhibitor, wortmannin, PKB inhibitor, API-2, and MAPK inhibitor, PD98059. These kinase inhibitors also prevented the ATP-induced increase in proliferation. In addition, ATP enhanced the progression of cells from the G0/G1 phase to the S phase by increasing the expression of proteins for cyclin D1 and cyclin E. Silencing the P2X4/7 and P2Y2 receptors with siRNA targeting the corresponding receptor diminished ATP-stimulated proliferation and migration of the cardiac fibroblasts. CONCLUSION AND IMPLICATION ATP activates P2X4/7 and P2Y2 receptors and up-regulates the proliferation of human cardiac fibroblasts by promoting cell cycling progression. It also increases the migration of these cells. These effects of ATP may be involved in cardiac remodelling of injured hearts.