Abstract：
In crustaceans, the steroid hormone 20-hydroxyecdysone (20E) initiates molting, and the molting process is also regulated by energy metabolism. AMPK is an energy sensor and plays a critical role in systemic energy balance. Here, the regulatory mechanism in the interaction between 20E and AMPK was investigated in Chinese mitten crab, Eriocheir sinensis. The results showed that the 20E concentration and the mRNA expression levels of 20E receptors in hepatopancreas were down-regulated post AMPK activator (AICAR) treatment, and were up-regulated after AMPK inhibitor (Compound C) injection in crabs. Besides, the molt-inhibiting hormone (MIH) gene expression in eyestalk showed the opposite patterns in response to the AICAR and Compound C treatment, respectively. Further investigation found that there was a significant reduction in 20E concentration post PI3K inhibitor (LY294002) treatment, and the phosphorylation level of PI3K was increased in hepatopancreas after AMPK inhibitor injection. On the other hand, the positive regulation of PI3K-mediated activation of AMPK was also observed, the phosphorylation levels of AMPKα, AMPKβ and PI3K in hepatopancreas were significantly increased post 20E injection. In addition, the phosphorylation levels of AMPKα and AMPKβ induced by 20E were decreased after the injection of PI3K inhibitor. Taken together, these results suggest that the regulatory cross-talk between 20E and AMPK is likely to act through PI3K pathway in E. sinensis, which appeared to be helpful for a better understanding in molting regulation.
摘要：
在甲壳类动物中，类固醇激素20-羟基蜕皮激素（20E）启动蜕皮，蜕皮过程也受能量代谢的调节。AMPK是一种能量传感器，在系统能量平衡中起着至关重要的作用。这里,在中华绒螯蟹中研究了AMPK与20E相互作用的调控机制，中华绒螯蟹。结果表明，AMPK激活剂（AICAR）处理后，肝胰腺中20E浓度和20E受体的mRNA表达水平下调，并且在螃蟹中注射AMPK抑制剂(化合物C)后上调。此外,眼柄中的蜕皮抑制激素（MIH）基因表达显示出相反的模式，以响应AICAR和化合物C处理，分别。进一步调查发现，PI3K抑制剂（LY294002）治疗后20E浓度显着降低，AMPK抑制剂注射后，肝胰腺中PI3K的磷酸化水平升高。另一方面,还观察到PI3K介导的AMPK激活的正调节，AMPKα的磷酸化水平，肝胰腺中的AMPKβ和PI3K在20E注射后显著增加。此外,注射PI3K抑制剂后，20E诱导的AMPKα和AMPKβ的磷酸化水平降低。一起来看,这些结果表明，20E和AMPK之间的调节交叉对话可能通过PI3K途径在中华大肠杆菌中发挥作用，这似乎有助于更好地理解蜕皮法规。
