哌可酸盐和牛磺酸是大鼠赖氨酸和苏氨酸缺乏的尿液生物标志物。

Moro Joanna, Roisné-Hamelin Gaëtan, Khodorova Nadezda, Rutledge Douglas N, Martin Jean-Charles, Barbillon Pierre, Tomé Daniel, Gaudichon Claire, Tardivel Catherine, Jouan-Rimbaud Bouveresse Delphine, Azzout-Marniche Dalila

Université Paris-Saclay, AgroParisTech, Institut National de recherche pour l'agriculture, l'alimentation et l'environnement, UMR Physiologie de la Nutrition et du Comportement Alimentaire, Palaiseau, France.

AgroParisTech, Université Paris-Saclay, Institut National de recherche pour l'agriculture, l'alimentation et l'environnement, UMR SayFood, Massy, France.

J Nutr. 2023 Sep;153(9):2571-2584. doi: 10.1016/j.tjnut.2023.06.039. Epub 2023 Jun 30.

BACKGROUND

The consumption of poor-quality protein increases the risk of essential amino acid (EAA) deficiency, particularly for lysine and threonine. Thus, it is necessary to be able to detect easily EAA deficiency.

OBJECTIVES

The purpose of this study was to develop metabolomic approaches to identify specific biomarkers for an EAA deficiency, such as lysine and threonine.

METHODS

Three experiments were performed on growing rats. In experiment 1, rats were fed for 3 weeks with lysine (L30), or threonine (T53)-deficient gluten diets, or nondeficient gluten diet (LT100) in comparison with the control diet (milk protein, PLT). In experiments 2a and 2b, rats were fed at different concentrations of lysine (L) or threonine (T) deficiency: L/T15, L/T25, L/T40, L/T60, L/T75, P20, L/T100 and L/T170. Twenty-four-hour urine and blood samples from portal vein and vena cava were analyzed using LC-MS. Data from experiment 1 were analyzed by untargeted metabolomic and Independent Component - Discriminant Analysis (ICDA) and data from experiments 2a and 2b by targeted metabolomic and a quantitative Partial Least- Squares (PLS) regression model. Each metabolite identified as significant by PLS or ICDA was then tested by 1-way ANOVA to evaluate the diet effect. A two-phase linear regression analysis was used to determine lysine and threonine requirements.

RESULTS

ICDA and PLS found molecules that discriminated between the different diets. A common metabolite, the pipecolate, was identified in experiments 1 and 2a, confirming that it could be specific to lysine deficiency. Another metabolite, taurine, was found in experiments 1 and 2b, so probably specific to threonine deficiency. Pipecolate or taurine breakpoints obtained give a value closed to the values obtained by growth indicators.

CONCLUSIONS

Our results showed that the EAA deficiencies influenced the metabolome. Specific urinary biomarkers identified could be easily applied to detect EAA deficiency and to determine which AA is deficient.

背景

低质量蛋白质的摄入会增加必需氨基酸（EAA）缺乏的风险，尤其是赖氨酸和苏氨酸。因此，有必要能够轻松检测EAA缺乏情况。

目的

本研究的目的是开发代谢组学方法，以鉴定EAA缺乏（如赖氨酸和苏氨酸缺乏）的特定生物标志物。

方法

对生长中的大鼠进行了三项实验。在实验1中，与对照饮食（乳蛋白，PLT）相比，给大鼠喂食赖氨酸（L30）或苏氨酸（T53）缺乏的面筋饮食或非缺乏面筋饮食（LT100）3周。在实验2a和2b中，给大鼠喂食不同浓度的赖氨酸（L）或苏氨酸（T）缺乏饮食：L/T15、L/T25、L/T40、L/T60、L/T75、P20、L/T100和L/T170。使用液相色谱-质谱联用仪分析来自门静脉和腔静脉的24小时尿液和血液样本。实验1的数据通过非靶向代谢组学和独立成分-判别分析（ICDA）进行分析，实验2a和2b的数据通过靶向代谢组学和定量偏最小二乘（PLS）回归模型进行分析。然后，对通过PLS或ICDA鉴定为显著的每种代谢物进行单因素方差分析，以评估饮食效果。采用两阶段线性回归分析来确定赖氨酸和苏氨酸的需求量。