Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Institute of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Henan Provincial Animal Husbandry General Station, Zhengzhou 450008, China.
Oxid Med Cell Longev. 2019 Nov 19;2019:1076512. doi: 10.1155/2019/1076512. eCollection 2019.
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 HO-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 HO-induced GC apoptosis. Furthermore, the overexpression of let-7a reduced the upregulation of Fas expression in HO-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 HO. These results indicate that GSPB2 inhibits HO-induced apoptosis of GCs, possibly through the upregulation of let-7a.
氧化应激是与颗粒细胞（GC）凋亡相关的各种生殖障碍的因果因素和关键促进因素，它通过调节相关基因的表达来发挥作用。各种研究表明，葡萄籽原花青素 B2（GSPB2）可能通过保护 GC 免受氧化损伤，尽管其潜在机制尚未完全阐明。因此，本研究探讨了 GSPB2 的有益作用是否与 microRNAs 有关，microRNAs 被认为通过调节蛋白质编码基因的表达在 GC 凋亡中发挥关键作用。结果表明，GSPB2 处理可通过 MTT 测定和 TUNEL 染色检测到保护 GC 免受 HO 诱导的凋亡，并增加 GC 中的 let-7a 表达。此外，let-7a 的过表达显着增加细胞活力并抑制 HO 诱导的 GC 凋亡。此外，let-7a 的过表达降低了 HO 处理的 GC 中 Fas 表达在 mRNA 和蛋白质水平的上调。双荧光素酶报告基因检测结果表明，let-7a 可直接靶向 Fas 3'-UTR。此外，let-7a 的过表达增强了 GSPB2 对 HO 诱导的 GC 凋亡的保护作用。这些结果表明，GSPB2 通过上调 let-7a 抑制 HO 诱导的 GC 凋亡。
