摘要： 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0.05,0.10 μg/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,0.05,0.10 μg/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,0.05,0.10 μg/mL剂量组P-AMPK的表达量均达到最高值(P<0.01),且0.10 μg/mL剂量组表达量明显高于0.05 μg/mL剂量组;食欲素A 刺激细胞2 h,0.05,0.10 μg/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,0.10 μg/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。
关键词:
食欲素 A,
颗粒细胞,
P-AMPK,
P-ERK
Abstract: 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 0.05,0.10 μg/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 μg/mL of orexins A 2 h treatment were significantly upregulated compared with that of control (P<0.01),respectively.The expression of P-ERK of the cells treated with 0.10 μg/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.
Key words:
Orexin A,
Granulosa cells,
P-AMPK,
P-ERK
中图分类号:
王伟平, 徐晓静, 杜晨光, 马春燕, 米焱, 刘永志. 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[J]. 华北农学报, 2015, 30(3): 136-139. doi: 10.7668/hbnxb.2015.03.024.
WANG Wei-ping, XU Xiao-jing, DU Chen-guang, MA Chun-yan, MI Yan, LIU Yong-zhi. Effect of Orexin A on the Expression of P-AMPK and P-ERK in Sheep Ovarian Granulosa Cells[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(3): 136-139. doi: 10.7668/hbnxb.2015.03.024.