Exploration of the mechanism of PCOS induced by microenvironmental changes in follicular fluid based on 16 S rRNA and metabolomics

基于16S rRNA和代谢组学的卵泡液微环境变化诱导多囊卵巢综合征机制研究

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Abstract

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder in women of childbearing age. The follicular microenvironment plays a vital role in oocyte development as one of the important factors affecting PCOS. This study aimed to reveal the changes in the follicular microenvironment of PCOS rats using multi-omics analysis. METHODS: A PCOS rat model was constructed using dehydroepiandrosterone (DHEA) method, and 16 S rRNA amplicon sequencing and non-targeted metabolomics were applied to analyze the follicular fluid samples from the control and the PCOS groups. The key microbiota were screened using T-test analysis, and the key metabolites were identified through Spearman correlation hierarchical cluster analysis. Bioinformatics and network pharmacology were used to identify overlapping genes between the key metabolite targets and PCOS-related targets, followed by Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. RESULTS: There were significant differences in the microbiome composition between the two groups, with a general decrease in the microbial abundance in the follicular fluid of the PCOS group compared to the control group. T-test analysis identified Acinetobacter haemolyticus as a significantly different strain. Spearman correlation analysis exhibited a positive correlation between Acinetobacter haemolyticus and three metabolites (S-adenosylmethioninamine, prorocentrolide, and cilostazol). Network pharmacology and bioinformatics analyses revealed that the overlapping genes of these metabolites targets and PCOS-related targets were enriched in autophagy-related signaling pathways, with cilostazol as a candidate metabolite and SRC as a potential target. Additionally, Liquid Chromatography-Mass Spectrometry (LC-MS) analysis confirmed the presence of Acinetobacter haemolyticus in the follicular fluid of rats and its ability to metabolize cilostazol. CONCLUSIONS: Cilostazol is a significantly differentiated metabolite in the follicular microenvironment of PCOS rats, playing a role in PCOS development by regulating autophagy-related signaling processes mediated by SRC. CLINICAL TRIAL NUMBER: Not applicable.

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