Integrated Analysis of miRNA-mRNA Interaction Network in Porcine Granulosa Cells Undergoing Oxidative Stress

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.

MicroRNA Expression Profiling in Porcine Liver, Jejunum and Serum upon Dietary DON Exposure Reveals Candidate Toxicity Biomarkers

“…Notably, toxic effects of DON comprise induction of oxidative stress and reactive oxygen species [ 30 , 31 , 32 ]. In the same line, high levels of mirR-128, as we observed in the present study in DON exposed pigs, have also been associated with higher oxidative stress [ 33 ].…”

Transcriptomic Data Analyses Reveal That Sow Fertility-Related lincRNA NORFA Is Essential for the Normal States and Functions of Granulosa Cells

“…Before the preparation for RNA-seq, the transfection efficiency and knockdown efficiency of siNORFA in porcine GCs were detected and validated. Subsequently, the cDNA libraries for sequencing were prepared as previously described ( Du et al, 2019 ). Briefly, after removing rRNA, total RNA was broken into 100- to 150-nt fragments and cDNA was synthesized and purified by using QIAquick PCR Purification kit (#28104, QIAGEN), which was subsequently sent to Biomarker Technologies Co. Ltd. (Beijing, China) for sequencing.…”

“…To construct the effective NORFA -mediated PPI network, all the function-known DEmRNA-encoded proteins were selected, and the network was established as previously described ( Du et al, 2019 ). Briefly, all the interactions (validated and predicted) between DEG-encoded proteins were analyzed and generated by using STRING v11.0 online statistical analysis software 4 ( Szklarczyk et al, 2019 ) with the basic settings: the interacted protein amount ≥1 and the minimum required interaction score ≥0.7 [0–1].…”

miR-31-5p Promotes Oxidative Stress and Vascular Smooth Muscle Cell Migration in Spontaneously Hypertensive Rats via Inhibiting FNDC5 Expression

“…MicroRNAs (miRNAs) are a class of endogenous, non-coding small RNAs (about 18-22 nucleotides) that negatively regulate gene expression by binding to the mRNAs of target genes, specifically on a posttranscriptional level [19]. The dysregulation of miRNAs contributes to the pathogenesis of oxidative stress [20,21], inflammation [22], and several diseases such as cancer [23] and vascular diseases [24,25]. miR-31-5p inhibits the cell migration of renal cell carcinoma [26] and hepatocellular carcinoma [27], but it promotes cell migration in colorectal cancer cells [28].…”