Peron Gregorio, Uddin Jalal, Stocchero Matteo, Mammi Stefano, Schievano Elisabetta, Dall'Acqua Stefano
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Via Marzolo 5, 35131 Padova, Italy.
Department of Chemical Sciences, University of Padova, Via Marzolo 1, 35131 Padova, Italy.
J Pharm Biomed Anal. 2017 Jun 5;140:62-70. doi: 10.1016/j.jpba.2017.03.015. Epub 2017 Mar 16.
A longitudinal study was performed to evaluate the effects of Polygonum cuspidatum extract (standardized at 20% resveratrol) supplementation on healthy rats. The effects were explored by monitoring urinary metabolome changes using UPLC-HRMS and H NMR-based approaches. The aim of the study was to explore the effects of P. cuspidatum supplementation on a healthy animal model using metabolomics, in order to determine possible modes of action and obtain information on bioactivity.
Healthy Sprague-Dawley rats were orally supplemented with 100mg/kg of dried P. cuspidatum extract for 49days and 24-h urinary outputs were collected. Samples were analysed by untargeted UPLC-HRMS and H NMR approaches and the obtained data sets were modelled by an adaptation of post-transformation of PLS2 to longitudinal studies. Putative markers were discovered by a stability selection procedure and specific oxidative stress markers were monitored by a targeted HPLC-MS/MS analysis to assess the in vivo antioxidant activity of P. cuspidatum extract.
UPLC-HRMS and H NMR platforms showed two different but complementary patterns of metabolites describing the changes ascribable to P. cuspidatum supplementation and using both approaches, a comprehensive resveratrol metabolism and urinary excretion could be observed. Markers of P. cuspidatum supplementation effects identified by UPLC-HRMS were mainly related to its antioxidant activity and to a possible "adaptogenic" activity. Urinary changes observed by H NMR were mainly related to energy metabolism. UPLC-HRMS and H NMR metabolomics approaches allowed the effects of a prolonged supplementation with P. cuspidatum on healthy rats to be observed. The statistical models built from both data sets showed metabolic changes in urines related to rat aging.
进行了一项纵向研究，以评估虎杖提取物（白藜芦醇标准化含量为20%）对健康大鼠的影响。通过使用超高效液相色谱-高分辨质谱联用仪（UPLC-HRMS）和基于核磁共振氢谱（H NMR）的方法监测尿液代谢组变化来探究这些影响。本研究的目的是利用代谢组学探究虎杖补充剂对健康动物模型的影响，以确定可能的作用模式并获取生物活性信息。
给健康的斯普拉格-道利大鼠口服100mg/kg的干燥虎杖提取物，持续49天，并收集24小时尿液输出量。通过非靶向UPLC-HRMS和H NMR方法分析样品，所得数据集通过将偏最小二乘判别分析（PLS2）的变换应用于纵向研究进行建模。通过稳定性选择程序发现推定标志物，并通过靶向高效液相色谱-串联质谱（HPLC-MS/MS）分析监测特定的氧化应激标志物，以评估虎杖提取物的体内抗氧化活性。
UPLC-HRMS和H NMR平台显示出两种不同但互补的代谢物模式，描述了虎杖补充剂引起的变化，使用这两种方法，可以观察到全面的白藜芦醇代谢和尿液排泄情况。通过UPLC-HRMS鉴定的虎杖补充剂效果标志物主要与其抗氧化活性和可能的“适应原性”活性有关。通过H NMR观察到的尿液变化主要与能量代谢有关。UPLC-HRMS和H NMR代谢组学方法能够观察到长期补充虎杖对健康大鼠的影响。从两个数据集建立的统计模型显示尿液中的代谢变化与大鼠衰老有关。
