Abstract
Oxidative stress is a causal factor and key promoter of all kinds of reproductive disorders related to granulosa cell (GC) apoptosis that acts by dysregulating the expression of related genes. Various studies have suggested that grape seed procyanidin B2 (GSPB2) may protect GCs from oxidative injury, though the underlying mechanisms are not fully understood. Therefore, whether the beneficial effects of GSPB2 are associated with microRNAs, which have been suggested to play a critical role in GC apoptosis by regulating the expression of protein-coding genes, was investigated in this study. The results showed that GSPB2 treatment protected GCs from a H2O2-induced apoptosis, as detected by an MTT assay and TUNEL staining, and increased let-7a expression in GCs. Furthermore, let-7a overexpression markedly increased cell viability and inhibited H2O2-induced GC apoptosis. Furthermore, the overexpression of let-7a reduced the upregulation of Fas expression in H2O2-treated GCs at the mRNA and protein levels. Dual-luciferase reporter assay results indicated that let-7a directly targets the Fas 3′-UTR. Furthermore, the overexpression of let-7a enhanced the protective effects of GSPB2 against GC apoptosis induced by H2O2. These results indicate that GSPB2 inhibits H2O2-induced apoptosis of GCs, possibly through the upregulation of let-7a.
Highlights
IntroductionNumerous follicles are present in both ovaries 10 days after birth in sows, more than 99% of follicles undergo a degenerative process known as atresia during growth and development [1]
grape seed procyanidin B2 (GSPB2) pretreatment significantly inhibited the H2O2-induced downregulation of let-7a expression in granulosa cell (GC) (P < 0:05). These results show that GSPB2 inhibits H2O2-induced GC apoptosis by regulating let-7a expression
Numerous studies have shown that GC apoptosis plays a key role in the regulation of ovarian follicular atresia [26, 34, 35]
Summary
Numerous follicles are present in both ovaries 10 days after birth in sows, more than 99% of follicles undergo a degenerative process known as atresia during growth and development [1]. Recent studies have indicated that granulosa cell (GC) apoptosis plays a crucial role in the initiation of this process [2, 3]. ROS are necessary for fundamental cellular processes, such as cell proliferation, follicle development, and ovulation [6]. Previous studies have indicated that ROS accumulation in the ovary can give rise to GC apoptosis and antral follicle atresia in rats [8]. Oxidative stress in vivo leads to GC apoptosis and follicular atresia [9]. The discovery and identification of an antioxidant by targeting oxidative stress-induced apoptosis may provide benefits for GC survival against oxidative injury
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