生物
胆固醇侧链裂解酶
细胞生物学
甾体生物合成
分解代谢
内科学
生物化学
内分泌学
新陈代谢
细胞色素P450
激素
类固醇
医学
作者
Manuela Hofner,Katja Eubler,Carola Herrmann,U. Berg,Dieter Berg,Harald Welter,Axel Imhof,Ignasi Forné,Artur Mayerhofer
出处
期刊:Molecular human reproduction
[Oxford University Press]
日期:2023-12-21
标识
DOI:10.1093/molehr/gaad049
摘要
Abstract 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 down-regulated. Pathways associated with glycolysis, chemical homeostasis, cellular response to hypoxia and actin filament bundle assembly were up-regulated. 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 (2-4 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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