氧气浓度降低会调节体外人类颗粒细胞的表型和功能,并导致类固醇生成减少,但炎症细胞反应增加。

IF 3.6 2区 医学 Q2 DEVELOPMENTAL BIOLOGY
Maria Höfner, Katja Eubler, Carola Herrmann, Ulrike Berg, Dieter Berg, Harald Welter, Axel Imhof, Ignasi Forné, Artur Mayerhofer
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引用次数: 0

摘要

氧气(O2)浓度作为卵巢细胞的重要调节因子最近引起了讨论。人类试管婴儿衍生的颗粒细胞(人类 GCs)可在体外维持,是一种广泛使用的人类卵巢细胞模型。通常情况下,GCs 是在大气氧气水平(约 20%)下培养的,而体内的氧气条件,尤其是排卵前卵泡中的氧气条件估计要低得多。因此,我们全面评估了在大气与低氧(1% O2)条件下培养 4 天对人类 GCs 的影响。我们发现,在1%氧气条件下,细胞RNA和蛋白质水平降低,但细胞数量不变,这表明转录和/或翻译活性降低。蛋白质组分析表明,在缺氧条件下,391种蛋白质含量确实降低了,但133种蛋白质含量却增加了。根据基因本体GO富集分析,与代谢过程相关的通路,如氨基酸代谢过程、线粒体蛋白质生物合成和类固醇生物合成,都出现了下调。与糖酵解、化学平衡、细胞对缺氧的反应和肌动蛋白丝束组装相关的途径则上调。随着 CYP11A1(一种胆固醇侧链裂解酶)水平的降低,孕酮的释放也随之减少。蛋白质组分析仪以及 IL-6 和 IL-8 酶联免疫吸附测定显示,缺氧导致促炎因子和血管生成因子分泌增加。免疫荧光研究显示,在急性(2-4 小时)暴露于 1%O2 时,低氧诱导因子 1α (HIF1α)在人 GCs 中核定位,而在暴露于 1%O2 4 天的细胞中则没有。因此,HIF1α的作用可能仅限于启动人类GCs的缺氧反应。这些结果提供了缺氧诱导的人类 GC 表型变化的详细情况,并揭示了长期低氧条件抑制了这些细胞的类固醇生成,但促进了其炎症表型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Reduced oxygen concentrations regulate the phenotype and function of human granulosa cells in vitro and cause a diminished steroidogenic but increased inflammatory cellular reaction.

Oxygen (O2) concentrations have recently been discussed as important regulators of ovarian cells. Human IVF-derived granulosa cells (human GCs) can be maintained in vitro and are a widely used cellular model for the human ovary. Typically, GCs are cultured at atmospheric O2 levels (approximately around 20%), yet the O2 conditions in vivo, especially in the preovulatory follicle, are estimated to be much lower. Therefore, we comprehensively evaluated the consequences of atmospheric versus hypoxic (1% O2) conditions for 4 days on human GCs. We found lower cellular RNA and protein levels but unchanged cell numbers at 1% O2, indicating reduced transcriptional and/or translational activity. A proteomic analysis showed that 391 proteins were indeed decreased, yet 133 proteins were increased under hypoxic conditions. According to gene ontology (GO) enrichment analysis, pathways associated with metabolic processes, for example amino acid-catabolic-processes, mitochondrial protein biosynthesis, and steroid biosynthesis, were downregulated. Pathways associated with glycolysis, chemical homeostasis, cellular response to hypoxia, and actin filament bundle assembly were upregulated. In accordance with lower CYP11A1 (a cholesterol side-chain cleavage enzyme) levels, progesterone release was decreased. A proteome profiler, as well as IL-6 and IL-8 ELISA assays, revealed that hypoxia led to increased secretion of pro-inflammatory and angiogenic factors. Immunofluorescence studies showed nuclear localization of hypoxia-inducible factor 1α (HIF1α) in human GCs upon acute (2 h) exposure to 1% O2 but not in cells exposed to 1% O2 for 4 days. Hence, the role of HIF1α may be restricted to initiation of the hypoxic response in human GCs. The results provide a detailed picture of hypoxia-induced phenotypic changes in human GCs and reveal that chronically low O2 conditions inhibit the steroidogenic but promote the inflammatory phenotype of these cells.

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来源期刊
Molecular human reproduction
Molecular human reproduction 生物-发育生物学
CiteScore
8.30
自引率
0.00%
发文量
37
审稿时长
6-12 weeks
期刊介绍: MHR publishes original research reports, commentaries and reviews on topics in the basic science of reproduction, including: reproductive tract physiology and pathology; gonad function and gametogenesis; fertilization; embryo development; implantation; and pregnancy and parturition. Irrespective of the study subject, research papers should have a mechanistic aspect.
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