Chien Peter Tzu-Yu, Lin Chih-Chung, Hsiao Li-Der, Yang Chuen-Mao
Graduate Institute of Biomedical Sciences, Health Ageing Research Center, College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan; Department of Physiology and Pharmacology, Health Ageing Research Center, College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan.
Department of Anesthetics, Chang Gung Memorial Hospital at Lin-Kou and College of Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan.
Mol Cell Endocrinol. 2015 Jul 5;409:59-72. doi: 10.1016/j.mce.2015.04.005. Epub 2015 Apr 11.
Thrombin and COX-2 regulating cardiac hypertrophy are via various signaling cascades. Several transcriptional factors including CREB involve in COX-2 expression. However, the interplay among thrombin, CREB, and COX-2 in primary human neonatal ventricular cardiomyocytes remains unclear. In this study, thrombin-induced COX-2 promoter activity, mRNA and protein expression, and PGE2 synthesis were attenuated by pretreatment with the inhibitors of c-Src (PP1), Pyk2 (PF431396), EGFR (AG1478), PI3K/Akt (LY294002/SH-5), and p300 (GR343), or transfection with siRNAs of c-Src, Pyk2, EGFR, p110, Akt, CREB, and p300. Moreover, thrombin-stimulated phosphorylation of c-Src, Pyk2, EGFR, Akt, CREB and p300 was attenuated by their respective inhibitors. These results indicate that thrombin-induced COX-2 expression is mediated through PAR-1/c-Src/Pyk2/EGFR/PI3K/Akt linking to CREB and p300 cascades. Functionally, thrombin-induced hypertrophy and ANF/BNP release were, at least in part, mediated through a PAR-1/COX-2-dependent pathway. We uncover the importance of COX-2 regarding human cardiomyocyte hypertrophy that will provide a therapeutic intervention in cardiovascular diseases.
凝血酶和COX-2通过各种信号级联反应调节心肌肥大。包括CREB在内的几种转录因子参与COX-2的表达。然而，在原代人新生儿心室心肌细胞中，凝血酶、CREB和COX-2之间的相互作用仍不清楚。在本研究中，用c-Src抑制剂（PP1）、Pyk2抑制剂（PF431396）、EGFR抑制剂（AG1478）、PI3K/Akt抑制剂（LY294002/SH-5）和p300抑制剂（GR343）预处理，或转染c-Src、Pyk2、EGFR、p110、Akt、CREB和p300的siRNA，可减弱凝血酶诱导的COX-2启动子活性、mRNA和蛋白表达以及PGE2合成。此外，凝血酶刺激的c-Src、Pyk2、EGFR、Akt、CREB和p300的磷酸化被其各自的抑制剂减弱。这些结果表明，凝血酶诱导的COX-2表达是通过PAR-1/c-Src/Pyk2/EGFR/PI3K/Akt与CREB和p300级联反应介导的。在功能上，凝血酶诱导的肥大和ANF/BNP释放至少部分是通过PAR-1/COX-2依赖性途径介导的。我们揭示了COX-2在人类心肌细胞肥大中的重要性，这将为心血管疾病提供一种治疗干预措施。
