Chalmers Jennifer A, Lin Shuo-Yen J, Martino Tami A, Arab Sara, Liu Peter, Husain Mansoor, Sole Michael J, Belsham Denise D
Department of Physiology, University of Toronto, Medical Sciences Building 3247A, 1 King's College Circle, Toronto, Ontario, M5S 1A8 Canada.
J Mol Endocrinol. 2008 Sep;41(3):117-24. doi: 10.1677/JME-08-0050. Epub 2008 Jun 12.
Neuroendocrine peptides express biologic activity relevant to the cardiovascular system, including regulating heart rate and blood pressure, though little is known about the mechanisms involved. Here, we investigated neuroendocrine gene expression underlying diurnal physiology of the heart. We first used microarray and RT-PCR analysis and demonstrate the simultaneous expression of neuroendocrine genes in normal murine heart, including POMC, GnRH, neuropeptide Y, leptin receptor, GH-releasing hormone, cocaine- and amphetamine-regulated transcript, proglucagon, and galanin. We examined diurnal gene expression profiles, with cosinar bioinformatics to evaluate statistically significant rhythms. The POMC gene exhibits a day/night, circadian or diurnal, pattern of expression in heart, and we postulated that this may be important to cardiac growth and renewal. POMC diurnal gene rhythmicity is altered in pressure-overload cardiac hypertrophy, when compared with control heart, and levels increased at the dark-to-light transition times. These findings are also consistent with the proposal that neuropeptides mediate adverse remodeling processes, such as occur in pathologic hypertrophy. To investigate cellular responses, we screened three cell lines representing fibroblasts, cardiac myocytes, and vascular smooth muscle cells (NIH3T3, heart line 1, and mouse vascular smooth muscle cell line 1 (Movas-1) respectively). POMC mRNA expression is the most notable in Movas-1 cells and, furthermore, exhibits rhythmicity with culture synchronization. Taken together, these results highlight the diverse neuroendocrine mRNA expression profiles in cardiovasculature, and provide a novel model vascular culture system to research the role these neuropeptides play in organ health, integrity, and disease.
神经内分泌肽具有与心血管系统相关的生物活性,包括调节心率和血压,但其涉及的机制尚不清楚。在此,我们研究了心脏昼夜生理背后的神经内分泌基因表达。我们首先使用微阵列和逆转录-聚合酶链反应分析,证明神经内分泌基因在正常小鼠心脏中同时表达,包括阿片促黑皮质素原(POMC)、促性腺激素释放激素(GnRH)、神经肽Y、瘦素受体、生长激素释放激素、可卡因和苯丙胺调节转录物、胰高血糖素原和甘丙肽。我们使用余弦生物信息学检查昼夜基因表达谱,以评估具有统计学意义的节律。POMC基因在心脏中呈现昼夜节律性表达模式,我们推测这可能对心脏生长和更新很重要。与对照心脏相比,压力超负荷性心肌肥大时POMC昼夜基因节律性发生改变,且在明暗交替时水平升高。这些发现也与神经肽介导不良重塑过程(如病理性肥大中发生的过程)的观点一致。为了研究细胞反应,我们筛选了分别代表成纤维细胞、心肌细胞和血管平滑肌细胞的三种细胞系(分别为NIH3T3、心脏系1和小鼠血管平滑肌细胞系1(Movas-1))。POMC mRNA表达在Movas-1细胞中最为显著,而且在培养同步时呈现节律性。综上所述,这些结果突出了心血管系统中多样的神经内分泌mRNA表达谱,并提供了一个新的血管培养系统模型来研究这些神经肽在器官健康、完整性和疾病中所起的作用。