不同浓度GnIH对鸭颗粒细胞周期和增殖的影响

为了研究不同浓度GnIH对鸭颗粒细胞周期、增殖及相关基因表达的影响.本研究分别用不同浓度GnIH(0、0.1、1、10和100 ng ? mL-1)处理体外培养的鸭颗粒细胞24 h(n=3),观察细胞的生长状态,通过流式细胞术和EdU方法检测细胞周期和细胞增殖,并用qRT-PCR方法检测增殖相关基因CDK6、Cycli...

Effects of Different Concentrations of GnIH on Cell Cycle and Proliferation of Duck Granulosa Cells

The study aimed to explore the effects of different concentrations of GnIH on cell cycle, proliferation and related gene expression of duck granulosa cells. Primary duck granulosa cells were cultured in vitro and treated with different concentrations of GnIH (0, 0.1, 1, 10 and 100 ng·mL^-1) for 24 h (n=3). The cell growth status was observed, and the cell cycle and proliferation were detected by flow cytometry and EdU method, and the expression of proliferation-related genes CDK6, CyclinD1, IGF-2, IGFBP-2, p27^kip1 were detected by qRT-PCR. The results showed that the growth status of the cells in the GnIH treatment group at each concentration was good, the morphology was normal, the cell outline was clear, and the number of dead cells were not significantly different between groups (P>0.05). In the 0.1 and 1 ng·mL^-1 GnIH treatment groups, the proportion of cells in the G2 phase increased significantly (P<0.05). With the increase of the concentration of GnIH, the percentage of EdU-positive cells were decreased. In the 0.1 and 1 ng·mL^-1 GnIH treatment groups, the relative expression of CDK6, CyclinD1, IGF-2, IGFBP-2, p27^kip1 genes were downregulated in granulosa cells. The relative expression of these genes were increased in the 10 and 100 ng·mL^-1 GnIH treatment groups. Studies have shown that in the duck granulosa cells cultured in vitro, GnIH could hinder the cell cycle in G2 phase, meanwhile the percentage of EdU-positive cells were decreased and the expre-ssion of proliferation-related genes were downregulated, which could inhibit the proliferation of granulosa cells to affect the reproductive performance of animals.