Laboratorio de Neurobiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile , Santiago Chile.
Cardio-Ventilatory Muscle Physiology Laboratory, Institute of Physiology and Pathophysiology, Heidelberg University Hospital , Heidelberg , Germany.
Physiol Genomics. 2019 Apr 1;51(4):109-124. doi: 10.1152/physiolgenomics.00114.2018. Epub 2019 Mar 1.
Sustained chronic hypoxia (CH) produces morphological and functional changes in the carotid body (CB). Nitric oxide (NO) and endothelin-1 (ET-1) play a major role as modulators of the CB oxygen chemosensory process. To characterize the effects of CH related to normoxia (Nx) on gene expression, particularly on ET-1 and NO pathways, primary cultures of rat CB cells were exposed to 7 days of CH. Total RNA was extracted, and cDNA-P was synthesized and hybridized with 1,185 genes printed on a nylon membrane Atlas cDNA Expression Array. Out of 324 differentially expressed genes, 184 genes were upregulated, while 140 genes were downregulated. The cluster annotation and protein network analyses showed that both NO and ET-1 signaling pathways were significantly enriched and key elements of each pathway were differentially expressed. Thus, we assessed the effect of CH at the protein level of nitric oxide synthase (NOS) isoforms and ET-1 receptors. CH induced an increase in the expression of endothelial NOS, inducible NOS, and ET. During CH, the administration of SNAP, a NO donor, upregulated ET. Treatment with Tezosentan (ET-1 receptor blocker) during CH upregulated all three NOS isoforms, while the NOS blocker L-NAME induced upregulation of iNOS and ET and downregulated the protein levels of ET. These results show that CH for 7 days changed the cultured cell CB gene expression profile, the NO and ET-1 signaling pathways were highly enriched, and these two signaling pathways interfered with the protein expression of each other.
慢性持续缺氧 (CH) 会导致颈动脉体 (CB) 发生形态和功能变化。一氧化氮 (NO) 和内皮素-1 (ET-1) 作为 CB 氧感受过程的调节剂发挥着重要作用。为了描述与常氧 (Nx) 相关的 CH 对基因表达的影响，特别是对 ET-1 和 NO 途径的影响，我们对大鼠 CB 细胞的原代培养物进行了 7 天的 CH 暴露。提取总 RNA，合成 cDNA-P，并与尼龙膜 Atlas cDNA Expression Array 上打印的 1,185 个基因进行杂交。在 324 个差异表达基因中，有 184 个基因上调，而 140 个基因下调。聚类注释和蛋白质网络分析表明，NO 和 ET-1 信号通路均显著富集，且每条通路的关键元素均表达不同。因此，我们评估了 CH 对一氧化氮合酶 (NOS) 同工型和 ET-1 受体蛋白水平的影响。CH 诱导内皮型 NOS、诱导型 NOS 和 ET 的表达增加。在 CH 期间，给予 NO 供体 SNAP 会使 ET 上调。在 CH 期间给予 Tezosentan（ET-1 受体阻滞剂）会使所有三种 NOS 同工型上调，而 NOS 阻滞剂 L-NAME 会诱导 iNOS 和 ET 的上调，并下调 ET 的蛋白水平。这些结果表明，7 天的 CH 改变了培养细胞 CB 的基因表达谱，NO 和 ET-1 信号通路高度富集，这两种信号通路相互干扰蛋白表达。
