亮氨酸对肉鸡肠上皮细胞TOR信号的调控研究.docVIP

亮氨酸对肉鸡肠上皮细胞TOR信号的调控研究 （王瑞贺 李然 丁显涛 韩静娅 史卫平） 指导老师：刘宁 所属领域：生命科学（农学） 摘要：【目的】本文旨在研究亮氨酸（Leu）对鸡肠上皮细胞（IEC）TOR基因mRNA表达的影响，探讨氨基酸调控蛋白质合成的作用机制，为建立精确营养供给技术提供理论基础。【方法】实验设置对照组和Leu处理组I（50mM）、II（100mM）、III（150mM）和IV（200mM），通过体外细胞培养和半定量RT-PCR技术，以β-actin为内参，根据TOR基因和内参基因mRNA扩增产物电泳带的光密度值，计算TOR基因mRNA相对表达量。【结果】实验组Leu表达量相对比值均显著高于对照组（P ﹤0.05）。4个实验组中，II组和III组显著高于I组和IV组（P ﹤0.05），但II和III组之间差异不显著（P ≥ 0.05）。【结论】Leu能显著提高鸡IEC中TOR基因表达，以100mM和150mM效果最好。 关键词：亮氨酸 肉鸡 肠上皮细胞 TOR信号 基因表达 Study on Regulation of TOR Gene by Leucine in Chicken Intestinal Epithelial Cell Abstract: The objective of this study was to investigate the effect of leucine（Leu）on the regulation of TOR gene mRNA expression in chicken intestinal epithelial cell（IEC）, for discussing the theory of amino acid regulation of protein synthesis and the establishment of the precise mechanism of nutrient supply technology. There were control group and experiments I, II, III and IV containing Leu at 0, 50, 100, 150, 200 mM, respectively. The relative values of TOR mRNA expressions were calculated based on the optical density of amplification product of TOR and β-actin using the methods of cell culture and semi-quantitative RT-PCR. The results showed that Leu upregulated the mRNA expression of TOR compared with control group（P﹤0.05）. The TOR expressions in groups II and III were higher than that in I and IV（P﹤0.05）, but there was not different between II and III（P ≥ 0.05）. The results suggest that Leu can increase the mRNA expression of TOR in chicken IEC, and the best effects in the present study are found in the concentrations of Leu at 100 and 150 mM. Key words：Leucine； Chicken；Intestinal Epithelial Cell；TOR Signal；Gene Express 1 引言 雷帕霉素靶蛋白（Target of Rapamycin，TOR）是一类进化上十分保守的Ser/Thr蛋白激酶，属于磷酸肌醇相关激酶家族成员，广泛存在于各种生物细胞中。TOR也是一种重要的信号分子，在调节细胞生长和细胞周期中起到中枢作用[1]。它能够激活下游信号传导通路，通过4E-BP1、S6K等翻译调节因子的磷酸化作用来传递外界营养状况、生长因子等信号，从而调节细胞内核糖体的发生、蛋白质的合成等生理过程，进而综合调控细胞的生长、增殖、凋亡和自噬[2]。TOR信号是营养、化学和运动等因素导致细胞生长和分化的一个关键信号，生长因子、营养素、能量、应激以及雷帕霉素（Rapamycin）等因素可以调控TOR信号活性[3