College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China.
College of Life Science, Hebei Agricultural University, Baoding 071000, China.
Int J Mol Sci. 2022 May 6;23(9):5183. doi: 10.3390/ijms23095183.
Short-term dietary supplementation of ewes during the luteal phase can increase fertility, most probably by stimulating glucose uptake by the follicles. However, the molecular mechanism of glucose regulation of follicular development has not yet been clarified, especially the further study of long non-coding RNA (lncRNA) in determining fertility during follicular development. We generated granulosa cell (GC) models of different doses of glucose (0, 2.1, 4.2, 8.4, 16.8 and 33.6 mM), and observed that the highest cell viability was recorded in the 8.4 mM group and the highest apoptosis rates were recorded in the 33.6 mM group. Therefore, a control group ( = 3, 0 mM glucose), a low glucose group ( = 3, add 8.4 mM glucose), and a high glucose group ( = 3, add 33.6 mM glucose) of GCs were created for next whole genomic RNA sequencing. In total, 18,172 novel lncRNAs and 510 annotated lncRNAs were identified in the GCs samples. Gene Ontology indicated that differentially expressed lncRNAs associated with cell apoptosis were highly enriched. Kyoto Encyclopedia of Genes and Genomes enrichment analysis of lncRNA target genes found that the apoptosis pathway and the p53 signaling pathway were both enriched. Furthermore, we focused on the function of a lnc and verified that lncG could influence cell apoptosis in GC development through affecting the mRNA and protein expression of apoptosis-related markers. These results provide the basis for further study of the lncRNA regulation mechanism in nutrition on female fertility.
短期在黄体期对母羊进行饮食补充可以提高繁殖力，这很可能是通过刺激卵泡摄取葡萄糖实现的。然而，卵泡发育过程中葡萄糖调节的分子机制尚不清楚，特别是在长链非编码 RNA（lncRNA）在决定卵泡发育过程中的生育能力方面的进一步研究。我们生成了不同葡萄糖剂量（0、2.1、4.2、8.4、16.8 和 33.6mM）的颗粒细胞（GC）模型，观察到细胞活力在 8.4mM 组中最高，凋亡率在 33.6mM 组中最高。因此，我们创建了对照组（=3，0mM 葡萄糖）、低糖组（=3，添加 8.4mM 葡萄糖）和高糖组（=3，添加 33.6mM 葡萄糖）的 GC 进行全基因组 RNA 测序。总共在 GC 样本中鉴定出 18172 个新的 lncRNA 和 510 个注释的 lncRNA。基因本体论表明，与细胞凋亡相关的差异表达 lncRNA 高度富集。lncRNA 靶基因京都基因与基因组百科全书富集分析发现，凋亡途径和 p53 信号通路都被富集。此外，我们还关注了一个 lnc 的功能，并通过影响与凋亡相关标志物的 mRNA 和蛋白表达验证了 lncG 可以影响 GC 发育中的细胞凋亡。这些结果为进一步研究营养对女性生育能力的 lncRNA 调控机制提供了依据。
