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Integrated Analysis of miRNA-mRNA Interaction Network in Porcine Granulosa Cells Undergoing Oxidative Stress
Xing Du1, Qiqi Li1, Qiuyu Cao1
1College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Abstract:
Oxidative stress (OS), a common intracellular phenomenon induced by excess reactive oxygen species (ROS) generation, has been shown to be associated with mammalian ovarian follicular development blockage and granulosa cell (GC) impairment. However, the mechanism involved in these effects remains unknown, and the effect of OS on the transcriptome profiles in porcine GCs has not been fully characterized. In this study, we found that hydrogen peroxide-mediated oxidative stress induced porcine GC apoptosis and impaired cell viability. Moreover, RNA-seq analysis showed that oxidative stress induced dramatic changes in gene expression in porcine GCs. A total of 2025 differentially expressed genes (DEGs) were identified, including 1940 DEmRNAs and 55 DEmiRNAs. Functional annotation showed that the DEGs were mainly associated with cell states and function regulation. In addition, multiple hub genes (FOXO1, SOD2, BMP2, DICER1, BCL2L11, FZD4, ssc-miR-424, and ssc-miR-27b) were identified by constructing protein-protein interaction and DEmiRNA-DEmRNA regulatory networks. Furthermore, a gene-pathway-function coregulatory network was established and demonstrated that these hub genes were enriched in FoxO, TGF-β, Wnt, PIK3-Akt, MAPK, and cAMP signaling pathways, which play important roles in regulating cell apoptosis, cell proliferation, stress responses, and hormone secretion. The current research provides a comprehensive perspective of the effects of oxidative stress on porcine GCs and also identifies potential therapeutic targets for oxidative stress-induced female infertility.
Insights
Oxidative stress harms ovarian granulosa cells (GCs) by inducing apoptosis and altering gene expression. This study identifies key genes and pathways involved, offering potential therapeutic targets for infertility.
Area of Science:
- Reproductive Biology
- Cellular Stress Response
- Genomics
Background:
- Oxidative stress (OS) from excess reactive oxygen species (ROS) impairs mammalian ovarian follicular development and granulosa cell (GC) function.
- The precise mechanisms and transcriptomic effects of OS on porcine GCs remain incompletely understood.
Purpose of the Study:
- To investigate the impact of hydrogen peroxide-induced oxidative stress on porcine granulosa cell viability, apoptosis, and gene expression profiles.
- To identify key genes, microRNAs, and signaling pathways regulated by oxidative stress in porcine GCs.
Main Methods:
- Porcine GCs were subjected to hydrogen peroxide-induced oxidative stress.
- RNA sequencing (RNA-seq) was employed to analyze differential gene expression (DEGs) and microRNA expression.
- Protein-protein interaction and regulatory networks were constructed to identify hub genes and pathways.
Main Results:
- Oxidative stress significantly reduced porcine GC viability and induced apoptosis.
- RNA-seq identified 2025 differentially expressed genes (1940 mRNAs, 55 miRNAs), primarily involved in cell state and function regulation.
- Hub genes (e.g., FOXO1, SOD2, BMP2) and miRNAs (e.g., ssc-miR-424) were identified, enriched in FoxO, TGF-β, Wnt, PI3K-Akt, MAPK, and cAMP signaling pathways.
Conclusions:
- Oxidative stress profoundly affects porcine GC transcriptome, impacting crucial cellular processes.
- Identified hub genes and pathways provide insights into OS-induced infertility mechanisms.
- These findings highlight potential therapeutic targets for mitigating oxidative stress-related female infertility.
