摘要
Vascular endothelial growth factor (VEGF) is crucial for follicle development through the regulation of granulosa cell (GC) function in some mammals, but its mechanism is unclear in yak (Bos grunniens). Therefore, the objectives of this study were to investigate the effects of VEGF on the viability, apoptosis and steroidogenesis of yak GCs. First, we investigated the localization of VEGF and its receptor (VEGFR2) in yak ovaries by immunohistochemistry analysis and evaluated the effect of culture medium containing different VEGF concentrations and culture times on the viability of yak GCs by Cell Counting Kit-8. Then, optimal treatment with 20 ng/mL VEGF for 24 h was selected to analyze the effects of this compound on intracellular reactive oxygen species levels by DCFH-DA kit, cell cycle and apoptosis by flow cytometry, steroidogenesis by ELISA kit and the expression of the related genes by RT‒qPCR. The results showed that VEGF and VEGFR2 were highly coexpressed in GCs and theca cells. GCs cultured in medium containing 20 ng/mL VEGF for 24 h significantly improved cell viability, decreased ROS production, promoted the transition from G1 phase to S phase (P < 0.05), increased the expression of the CCND1 (P < 0.05), CCNE1, CDK2, CDK4, and PCNA genes (P < 0.01) and decreased the expression of the P53 gene (P < 0.05). This treatment significantly reduced GC apoptosis (P < 0.05) by promoting the expression of BCL2 and GDF9 (P < 0.01) and inhibiting the expression of BAX and CASPASE3 (P < 0.05). VEGF promoted progesterone secretion (P < 0.05) accompanied by increased expression of HSD3B, StAR and CYP11A1 (P < 0.05). Taken together, our findings highlight the beneficial influence exerted by VEGF in improving GC viability and reducing ROS production and the apoptosis rate through the modulation of related gene expression.
摘要译文
血管内皮生长因子（VEGF）通过调节一些哺乳动物的颗粒细胞（GC）功能对于卵泡发育至关重要，但是在YAK（BOS Grunniens）中，其机制尚不清楚。因此，这项研究的目的是研究VEGF对YAK GC的生存力，凋亡和类固醇发生的影响。首先，我们通过免疫组织化学分析研究了VEGF及其受体（VEGFR2）在YAK卵巢中的定位，并评估了含有不同VEGF浓度和培养时间的培养基对细胞计数KIT-8的YAK GCS生存能力的培养基的影响。然后，选择20 ng/ml VEGF进行24小时的最佳处理，以分析该化合物通过DCFH-DA试剂盒，细胞周期和流式细胞术，ELISA KIT的类固醇生成以及ELISA KIT的类固醇生成以及的表达，分析该化合物对细胞内活性氧水平的影响。RT = QPCR的相关基因。结果表明，VEGF和VEGFR2在GC和theca细胞中高度共表达。在含有20 ng/ml VEGF的培养基中培养24小时的GCS显着提高了细胞活力，降低ROS产生，促进了从G1期到S相的过渡（P <0.05），增加了CCND1的表达（P <0.05），CCNE11，CCNE11，CDK2，CDK4和PCNA基因（P <0.01），并降低了p53基因的表达（p <0.05）。通过促进Bcl2和GDF9的表达（P <0.01）并抑制BAX和CASPASE3（P <0.05），这种治疗可显着降低GC凋亡（P <0.05）。VEGF促进孕酮分泌（P <0.05），伴随着HSD3B，Star和CYP11A1的表达增加（P <0.05）。综上所述，我们的发现突出了VEGF通过调节相关基因表达来提高GC生存能力和降低ROS产生和凋亡率的有益影响。
You-guo Tan [b]. Effects of vascular endothelial growth factor (VEGF) on the viability, apoptosis and steroidogenesis of yak (Bos grunniens) granulosa cells[J]. Theriogenology, 2023,207: 1-10