Table7_Identification of microRNAs implicated in modulating resveratrol-induced apoptosis in porcine granulosa cells.XLSX

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（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。本团队前期研究中构建了RSV处理POGCs的模型，证实了RSV对POGCs的调控作用。为探究RSV对POGCs的miRNA水平调控效应以筛选差异表达miRNA，本研究设置对照组（n=3，0 μM RSV组）、低剂量RSV组（n=3，50 μM RSV组）与高剂量RSV组（n=3，100 μM RSV组）进行小RNA测序。共计鉴定出113个差异表达miRNA（differentially expressed miRNAs, DE-miRNAs），RT-qPCR分析结果与测序数据具有良好相关性。功能注释分析显示，LOW vs. CON组的差异miRNA可能参与细胞发育、增殖及凋亡过程；HIGH vs. CON组中，RSV的生物学功能主要与代谢过程及应激响应相关，所富集的通路涉及PI3K24、Akt、Wnt及细胞凋亡。此外，本研究构建了与凋亡及代谢相关的miRNA-mRNA调控网络，并筛选出ssc-miR-34a与ssc-miR-143-5p作为核心miRNA。综上，本研究深化了对RSV通过miRNA调控POGCs凋亡效应的理解，结果表明RSV可能通过促进miRNA表达以诱导POGCs凋亡，为解析miRNA联合RSV在猪卵巢颗粒细胞发育中的作用提供了更为清晰的认知。