BACKGROUND AND PURPOSE Several members of the core clock mechanism are equipped with a Per-Arnt-Sim (PAS) domain through which they can bind haem [Fe(II)]. Haem is a ligand for the orphan receptors REV-ERBα/β (NR1D1/2), which regulate circadian rhythm and metabolism. The ability to bind haem sensitises these clock components to the action of small molecule gases, including NO, CO and H2S. Studies conducted with European hamsters revealed that during winter sleep, key clock genes stop oscillating. At the same time, H2S, when administered at subtoxic concentrations, can induce a hypometabolic state in the cell. We suppose that core clock components, including the nuclear receptors REV-ERBs, neuronal PAS domain protein 2 (nPAS2) and PER2, can be H2S targets. The general objective of this study was to investigate the effect of the H2S system on the expression profile of the core clock genes in cells in vitro. EXPERIMENTAL APPROACH We analysed the expression of Per1, Per2, Per3, Bmal1, Cry1, Cry2, Nr1d1, Nfil-3 and Dbp messenger RNA (mRNA) in serum-shocked NIH-3T3 cells treated with a slow-releasing H2S donor (GYY4137) or the cystathionine beta-synthase (CBS) inhibitor (AOAA) cultured under constant darkness and collected during 3 days in 3 h interval. KEY RESULTS AND CONCLUSIONS AND IMPLICATIONS We found that pharmacological CBS inhibition increased the general expression and dynamics of several clock genes. On the other hand, increased H2S decreased Per2 expression. These data suggest that CBS can affect circadian clock and effect on clock-controlled transcription output.

中文翻译：

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通过处理GYY4137或AOAA，改变了血清冲击的NIH-3T3细胞中时钟基因的昼夜节律振荡的动力学变化。

背景和目的核心时钟机制的几个成员都配备了Per-Arnt-Sim（PAS）域，通过它们可以绑定血红素[Fe（II）]。血红素是孤儿受体REV-ERBα/β（NR1D1 / 2）的配体，后者调节昼夜节律和新陈代谢。结合血红素的能力使这些时钟成分对小分子气体（包括NO，CO和H2S）的作用敏感。与欧洲仓鼠进行的研究表明，在冬季睡眠期间，关键的时钟基因停止振荡。同时，当以亚毒性浓度施用H2S时，它可以在细胞中诱导低代谢状态。我们假设核心时钟组件，包括核受体REV-ERB，神经元PAS域蛋白2（nPAS2）和PER2，可以成为H2S靶标。这项研究的总体目标是研究H2S系统对体外细胞中核心时钟基因表达谱的影响。实验方法我们分析了用缓释H2S供体（GYY4137）处理的血清冲击的NIH-3T3细胞中Per1，Per2，Per3，Bmal1，Cry1，Cry2，Nr1d1，Nfil-3和Dbp Messenger RNA（mRNA）的表达。或在恒定黑暗条件下培养并在3小时间隔内收集3天的胱硫醚β-合酶（CBS）抑制剂（AOAA）。关键结果，结论和意义我们发现，药理性CBS抑制作用增加了几种时钟基因的一般表达和动力学。另一方面，增加的H2S降低了Per2的表达。这些数据表明，CBS可以影响生物钟和对时钟控制的转录输出的影响。