Integrated analysis of lncRNA and mRNA for the apoptosis of porcine ovarian granulosa cells after polyphenol resveratrol treatment

Abstract

Resveratrol (RES) is a non-flavonoid polyphenol compound that can be involved in follicular development and ovulation. However, the mechanism by which resveratrol regulates the apoptosis of porcine ovarian granulosa cells (POGCs) through long non-coding RNA (lncRNA) is poorly understood. We generated POGCs models of different doses of RES (0, 25, 50, 75, and 100 μM). It was observed that the cell viability was the highest in the 50 μM group, and the highest apoptosis rates were recorded in the 100 μM group. Therefore, a control group (n = 3, 0 μM RES group), a low RES group (n = 3, 50 μM RES group), and a high RES group (n = 3, 100 μM RES group) of POGCs were created for next RNA sequencing. Gene Ontology (GO) indicated that differentially expressed lncRNAs associated with apoptotic process were highly enriched. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of lncRNA target genes found that the Wnt signaling pathway and PI3K-Akt signaling pathway were both enriched. Furthermore, we constructed lncRNA-mRNA networks related to Metabolic and Cell Apoptosis, respectively. In the networks, five key-lncRNAs were screened, which may play a significant role in the process of POGCs metabolism and apoptosis. Furthermore, we focused on the function of a lnc-GAM (lncRNA associated with Granulosa cells Apoptosis and Metabolism) and verified that lnc-GAM could influence cell apoptosis in POGCs development by affecting the mRNA expression of apoptosis-related markers, and also affects the secretion of steroid hormones and related genes expression in POGCs cultured in vitro. Our study provides seminal data and important new insights into the regulation of reproductive mechanisms in porcine and other female mammals.

Identification of microRNAs implicated in modulating resveratrol-induced apoptosis in porcine granulosa cells

“…Then, the POGCs (pellet) were immediately seeded in 6-well plates and cultured (10 6 viable cells/well) in a humidified atmosphere (5% CO 2 , 95% air, 37°C) in Dulbecco’s modified Eagle’s medium supplemented (Invitrogen, Carlsbad, CA, United States) with 10% fetal bovine serum and 1% penicillin/streptomycin mixture (Invitrogen). When the POGCs reached 80% confluency, the medium was removed, and the cells were treated with 0, 50 and 100 μM of RSV (Solarbio, Beijing, China; Zhang et al, 2023 ), representing control, LOW and HIGH group, respectively. The POGCs were cultured with treatments in triplicate for 24 h under conditions described above.…”

“…Our previous study focused on lncRNA-mRNA changes in porcine ovarian granulosa cells (POGCs) after RSV treatment and was conducted to identify the differentially expressed lncRNAs and mRNAs ( Zhang et al, 2023 ); however, it is unknown whether miRNAs changed in POGCs after RSV treatment. Therefore, this study mainly focuses on the identification of the specific miRNAs influenced by RSV treatment on POGCs apoptosis, which aims to investigate the roles of RSV in including miRNA for the better understanding, and to reveal new candidate regulatory miRNAs and genes underlying the apoptosis of POGCs.…”

Resveratrol Promotes Proliferation, Antioxidant Properties, and Progesterone Production in Yak (Bos grunniens) Granulosa Cells

“…The concentrations of RES utilized were derived from prior studies on its supplementation in pig and sheep GCs [34,35]. The cell viability was determined using a CCK-8 assay in 96-well plates (5 × 10 3 cells/well).…”

“…Recent studies have shown that RES plays an important role in regulating animal reproduction [20]. The effects of RES on GCs have been studied in many mammalian species [33][34][35], but the role of RES in yak GCs is unclear. Hence, the aims of this study were to investigate the impact of resveratrol (RES) on several aspects of yak granulosa cells (GCs), including their viability, proliferation, apoptosis, cell cycle, antioxidant characteristics, lipid droplet concentration, ATP generation, steroid hormone secretion, and associated gene expression.…”

The effects of natural products and bioactive ingredients of traditional Chinese medicine on apoptosis of ovarian granulosa cells

“…A recent study demonstrated that apigenin and rutin inhibited cell apoptosis and promoted cell proliferation in healthy porcine GCs (Sirotkin, Záhoranska, et al, 2020). Icaritin and icariin are main chemical components of Epimedii herba , which were demonstrated to attenuate GC apoptosis and remarkably reduce expression of Bax and increase that of Bcl‐2 (Shao & Wang, 2021; Zhang et al, 2017). Daidzein was shown to inhibit yak GC apoptosis by activating mitochondrial apoptosis pathway, with an upregulation of AKT1, Bcl‐2, and P53 and downregulation of Bad (Bcl‐2 associated death) and Bax (Ayimuguli et al, 2022).…”