Table8_Identification of microRNAs implicated in modulating resveratrol-induced apoptosis in porcine granulosa cells.XLS

MicroRNAs (miRNAs) are small, noncoding RNAs that play a crucial role in the complex and dynamic network that regulates the apoptosis of porcine ovarian granulosa cells (POGCs). Resveratrol (RSV) is a nonflavonoid polyphenol compound that is involved in follicular development and ovulation. In previous study, we established a model of RSV treatment of POGCs, confirming the regulatory effect of RSV in POGCs. To investigate the miRNA-level effects of RSV on POGCs to reveal differentially expressed miRNAs, a control group (n = 3, 0 μM RSV group), a low RSV group (n = 3, 50 μM RSV group), and a high RSV group (n = 3, 100 μM RSV group) were created for small RNA-seq. In total, 113 differentially expressed miRNAs (DE-miRNAs) were identified, and a RT-qPCR analysis showed a correlation with the sequencing data. Functional annotation analysis revealed that DE-miRNAs in the LOW vs. CON group may be involved in cell development, proliferation, and apoptosis. In the HIGH vs. CON group, RSV functions were associated with metabolic processes and responses to stimuli, while the pathways were related to PI3K24, Akt, Wnt, and apoptosis. In addition, we constructed miRNA-mRNA networks related to Apoptosis and Metabolism. Then, ssc-miR-34a and ssc-miR-143-5p were selected as key miRNAs. In conclusion, this study provided an improved understanding of effects of RSV on POGCs apoptosis through the miRNA modulations. The results suggest that RSV may promote POGCs apoptosis by stimulating the miRNA expressions and provided a better understanding of the role of miRNAs combined with RSV in ovarian granulosa cell development in pigs.

微小RNA（MicroRNAs, miRNAs）是一类小型非编码RNA，在调控猪卵巢颗粒细胞（porcine ovarian granulosa cells, POGCs）凋亡的复杂动态网络中发挥关键作用。白藜芦醇（Resveratrol, RSV）是一种非黄酮类多酚化合物，参与卵泡发育与排卵过程。本团队前期研究中，我们构建了RSV处理猪卵巢颗粒细胞的模型，证实了RSV对猪卵巢颗粒细胞的调控效应。为探究RSV对猪卵巢颗粒细胞的微小RNA层面调控作用，筛选差异表达微小RNA（differentially expressed miRNAs, DE-miRNAs），我们设置三组样本开展小RNA测序：对照组（n=3，0μM RSV组）、低剂量RSV组（n=3，50μM RSV组）与高剂量RSV组（n=3，100μM RSV组）。本研究共鉴定得到113个DE-miRNAs，RT-qPCR验证结果与测序数据具有良好相关性。功能注释分析显示，低剂量组vs对照组（LOW vs. CON）中的DE-miRNAs可能参与细胞发育、增殖及凋亡过程；高剂量组vs对照组（HIGH vs. CON）中，RSV的调控功能与代谢过程及应激响应密切相关，涉及的信号通路包括PI3K24、Akt、Wnt及凋亡通路。此外，我们构建了凋亡与代谢相关的miRNA-mRNA调控网络，并筛选得到ssc-miR-34a与ssc-miR-143-5p作为核心微小RNA。综上，本研究加深了对RSV通过调控微小RNA影响猪卵巢颗粒细胞凋亡的认知，结果表明RSV可能通过激活微小RNA的表达促进猪卵巢颗粒细胞凋亡，同时为解析联合RSV与微小RNA在猪卵巢颗粒细胞发育中的作用提供了更深入的理论依据。