基于AMPK信号通路探究大豆抗原蛋白对鲤鱼肠黏膜能量代谢影响及调控的分子机制

大豆抗原蛋白glycinin和β-conglycinin是损害肠道健康的主要抗营养因子，前期研究表明α-酮戊二酸(AKG)可以促进肠道发育，是否与肠黏膜能量代谢有关及作用机制还不清楚。本项目拟通过正常生理状态、AMPK强化剂和抑制剂，同时结合肠道上皮细胞培养，利用RT-PCR、Western blot和代谢组学等技术，探究glycinin和β-conglycinin对鲤肠黏膜能量代谢、代谢组变化规律和肠道健康因子（紧密连接蛋白、细胞凋亡信号、炎性细胞因子）的影响，分析能量代谢与肠道健康因子的关系，挖掘能量代谢调节通路AMPK的作用与机制；阐明AKG对肠黏膜能量代谢和肠道健康的作用机理；探究AKG通过AMPK信号通路调控鲤肠粘膜能量代谢降低glycinin和β-conglycinin的损害及机制。本项目从肠黏膜能量代谢这一新的视角研究大豆蛋白利用与肠道健康，这对提高其应用具有重要的理论价值。

Soybean antigen glycinin and β-conglycinin are the main anti-nutritional factors that damage intestinal health, previous studies showed that α-ketoglutaric acid(AKG) could improve intestinal development, it is not clear whether it is related to the energy metabolism of intestinal mucosa and its mechanism. This project is intended to combine the normal physiological state, AMPK enhancer and inhibitor, and intestinal epithelial cell culture, using technologies such as RT-PCR, Western blot and metabonomics，investigate the effects of glycinin and β-conglycinin on energy metabolism, metabonomics and intestinal health factors (tight junction protein, apoptosis signal, inflammatory cytokines) in intestinal mucosa of common carp, analyses the relationship between energy metabolism and intestinal health factors, study the role and mechanism of AMPK in energy metabolism regulation pathway; elucidate the mechanism of AKG on intestinal mucosal energy metabolism and intestinal health; explore the mechanism of AKG regulation of energy metabolism in intestinal mucosa of carp through AMPK signaling pathway to decrease glycinin and β-conglycinin damage. This project studies soybean protein utilization and intestinal health from the perspective of intestinal mucosal energy metabolism, which has important theoretical value for improving its application.