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／mL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／mL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／mL剂量组P-AMPK的表达量均达到最高值（P＜0．01），且0．10μg／mL剂量组表达量明显高于0．05μg／mL剂量组；食欲素A刺激细胞2 h，0．05，0．10μg／mL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／mL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 0.10μg/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 0.05,0.10μg/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 0.10μg/mL of orexins A 24 h treatment were significantly higher than that of the plateau phase (6-12 h) and reached the extreme value(P<0.01).It was indicated that the orexins A was involved in the regulation of reproductive function by regulating the energy metabolism , growth and development of sheep throughout the signaling pathway of P-AMPK and P-ERK.
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