食欲素A对绵羊卵巢颗粒细胞P-AMPK与P-ERK信号通路的影响

Effect of Orexin A on the Expression of P-AMPK and P-ERK in Sheep Ovarian Granulosa Cells

为研究食欲素A对绵羊卵巢能量代谢及生长发育的调节作用,在体外培养黄体化颗粒细胞,用浓度为0.05,0.10μg/m L的食欲素A刺激细胞后,分别于0,2,6,12,24 h提取细胞总蛋白,采用Western Blot法对P-AMPK及P-ERK的表达量进行检测。结果显示,食欲素A刺激细胞2 h,0.05,0.10μg/m L剂量组的P-AMPK表达量与对照组相比均极显著提高（P〈0.01）,食欲素A刺激细胞6 h,0.05,0.10μg/m L剂量组P-AMPK的表达量均达到最高值（P〈0.01）,且0.10μg/m L剂量组表达量明显高于0.05μg/m L剂量组;食欲素A刺激细胞2 h,0.05,0.10μg/m L剂量组的P-ERK表达量与对照组相比均极显著提高（P〈0.01）,食欲素A刺激细胞24 h,0.10μg/m L剂量组的P-ERK表达量极显著高于平台期（6-12 h）（P〈0.01）并达到最高值。表明食欲素A通过P-AMPK和P-ERK途径参与黄体能量代谢及生长发育的调节,从而进一步参与生殖机能的调控。

In order to investigate the effect of orexins A on the energy metabolism,growth and development of sheep ovary,the luteinising granulosa cells were isolated and cultured in vitro. The passage luteinising granulosa cells were treated with 0.05,0.10 μg/mL of orexins A, and the total proteins of the cells were extracted at 0,2,6, 12,24 h treatment,respectively. The relative expression of P-AMPK and P-ERK were determined by Western Blot. The results showed that the expression of P-AMPK of the cells treated with 0.05,0.10μg/mL of orexins A 2 h after treatment were significantly upregulated compared with that of control （ P 〈 0.01 ） , respectively. The expression of P-AMPK of the cells treated with 0.05,0.10 μg/mL of orexins A 6 h treatment reached the extreme value （P 〈 0.01 ） , respectively, and expression of P-AMPK of the cells treated with 0.10 μg/mL of orexins A were significantly higher than that of the 0.05 μg/mL treated group. The expression of P-ERK of the cells treated with 0.05,0.10 vg/mL of orexins A 2 h treatment were significantly upregulated compared with that of control （P 〈 0.01 ） , respec- tively. The expression of P-ERK of the cells treated with 0.10 μg/mL of orexins A 24 h treatment were significantlyhigher than that of the plateau phase （6 - 12 h） and reached the extre the orexins A was involved in the regulation of reproductive function b and development of sheep throughout the signaling pathway of P-AMPK me value（P 〈0.01 ）. It was indicated that y regulating and P-ERK energy metabolism, growth