Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, P. R. China.
Jiangsu Livestock Embryo Engineering Laboratory, Nanjing Agricultural University, Nanjing 210095, P. R. China; and National Experimental Teaching Demonstration Centre of Animal Science, Nanjing Agricultural University, Nanjing 210095, P. R. China; and Corresponding author. Email:
Reprod Fertil Dev. 2020 Feb;32(3):335-348. doi: 10.1071/RD18442.
The objective of this study was to investigate the dose-dependent effect of 1α,25-(OH)2VD3 (Vit D3) on invitro proliferation of goat luteinised granulosa cells (LGCs) and to determine the underlying mechanisms of its action by overexpressing and silencing vitamin D receptor (VDR) in LGCs. Results showed that VDR was prominently localised in GCs and theca cells (TCs) and its expression increased with follicle diameter, but was lower in atretic follicles than in healthy follicles. The proliferation rate of LGCs was significantly higher in the Vit D3-treated groups than in the control group, with the highest proliferation rate observed in the 10nM group; this was accompanied by changes in the expression of cell cycle-related genes. These data indicate that Vit D3 affects LGC proliferation in a dose-dependent manner. Contrary to the VDR knockdown effects, its overexpression upregulated and downregulated cell cycle- and apoptosis-related genes respectively; moreover, supplementation with 10nM of Vit D3 significantly enhanced these effects. These results suggest that changes in VDR expression patterns in LGCs may be associated with follicular development by regulation of cell proliferation and apoptosis. These findings will enhance the understanding of the roles of Vit D3 and VDR in goat ovarian follicular development.
本研究旨在探讨 1α,25-(OH)2VD3（维生素 D3）对体外山羊黄体化颗粒细胞（LGC）增殖的剂量依赖性影响，并通过在 LGC 中过表达和沉默维生素 D 受体（VDR）来确定其作用的潜在机制。结果表明，VDR 主要定位于 GC 和膜细胞（TC）中，其表达随卵泡直径的增加而增加，但在闭锁卵泡中低于健康卵泡。与对照组相比，Vit D3 处理组的 LGC 增殖率显著升高，其中 10nM 组的增殖率最高；这伴随着与细胞周期相关基因的表达变化。这些数据表明，Vit D3 以剂量依赖的方式影响 LGC 的增殖。与 VDR 敲低的作用相反，其过表达分别上调和下调细胞周期和凋亡相关基因；此外，补充 10nM 的 Vit D3 显著增强了这些作用。这些结果表明，LGC 中 VDR 表达模式的变化可能通过调节细胞增殖和凋亡与卵泡发育有关。这些发现将增强对 Vit D3 和 VDR 在山羊卵巢卵泡发育中的作用的理解。
