Effects of N-Acetylcysteine on the Proliferation, Hormone Secretion Level, and Gene Expression Profiles of Goat Ovarian Granulosa Cells

Abstract

The purpose of this paper was to investigate the effects of N-acetylcysteine (NAC) on the proliferation, hormone secretion, and mRNA expression profiles of ovarian granulosa cells (GCs) in vitro. A total of 12 ovaries from 6 follicular-stage goats were collected for granulosa cell extraction. The optimum concentration of NAC addition was determined to be 200 μM via the Cell Counting Kit 8 (CCK-8) method. Next, GCs were cultured in a medium supplemented with 200 μM NAC (200 μM NAC group) and 0 μ M NAC (control group) for 48 h. The effects of 200 μM NAC on the proliferation of granulosa cells and hormones were studied by 5-ethynyl-2′-deoxyuridine (EdU) assay and enzyme-linked immunosorbent assay (ELISA). mRNA expression was analyzed by transcriptome sequencing. The results indicate that 200 μM NAC significantly increased cell viability and the proportion of cells in the S phase but promoted hormone secretion to a lesser degree. Overall, 122 differentially expressed genes (DEGs) were identified. A total of 51 upregulated and 71 downregulated genes were included. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that the most DEGs were enriched in terms of cell growth regulation, cell growth, neuroactive ligand-receptor interaction, cytokine-cytokine receptor interaction, the cAMP-signaling pathway, and the Wnt-signaling pathway. Seven genes related to granulosa cell proliferation were screened, IGFBP4, HTRA4, SST, SSTR1, WISP1, DAAM2, and RSPO2. The above results provide molecular theoretical support for NAC as a feed additive to improve follicle development and improve reproductive performance in ewes.

Continuous exposure to 60 Hz extremely low frequency electromagnetic field at 10 to 16 mT promotes various human cell proliferation by activating extracellular-signal-regulated kinase

“…A minor increase in intracellular ROS levels may induce cell proliferation by activating signaling cascades [23]. Antioxidants also increase cell proliferation by reducing intracellular ROS levels [24, 25]. Thus, we questioned whether the increased proliferation by 60 Hz ELF-EMF was triggered by changes in intracellular and/or mitochondrial ROS levels.…”

Assessment of the Effect of Leonurine Hydrochloride in a Mouse Model of PCOS by Gene Expression Profiling

“…Among these pathways, neuroactive ligand-receptor interactions were enriched in the highest number of DEGs and showed the most significant differences, followed by the calcium signalling pathway and the cAMP signalling pathway. Studies have shown that interactions between neuroactive ligands and receptors are involved in granulosa cell proliferation [43], and these interactions, along with signalling pathways such as the PPAR and cAMP signalling pathways, are important for ovarian development [44] and are related to the occurrence and development of PCOS [45,46]. The calcium signalling pathway is crucial in regulating various cellular processes such as cell growth, differentiation, proliferation, apoptosis, and other cellular processes [47].…”

N-Acetylcysteine Alleviates D-Galactose-Induced Injury of Ovarian Granulosa Cells in Female Rabbits by Regulating the PI3K/Akt/mTOR Signaling Pathway

“…In in vitro experiments, the addition of 1.5 mM of NAC to oocyte maturation medium has been shown to increase the proportion of embryos reaching the blastocyst stage [18]. Moreover, 200 µM of NAC could activate the cAMP and Wnt signaling pathways to significantly enhance proliferation and hormonal secretion by goat ovarian GCs, with the process being essential for improving follicular growth and reproductive performance in ewes [19]. Dietary supplementation of 0.07% NAC has further been found to regulate endometrial immune and inflammatory responses through the PI3K/Akt and PPAR pathways, hence improving the survival rate of goat embryos in early pregnancy [20].…”