A method for constructing a rat model for anti-oxidative aging drug screening based on metabolomics analysis

A rat model and aging technology, applied in the field of metabolomics analysis, can solve the problems of low accuracy, poor system, high cost, etc., and achieve the effect of strong specificity, embodying rationality and scientificity

Active Publication Date: 2019-08-20
WUHAN UNIV
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  • Application Information

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Problems solved by technology

[0012] In order to solve the technical problems of low accuracy, high cost, and poor system in the existing rat model construction and evaluation methods for anti-oxidative aging drug screening, the present invention provides an anti-oxidative aging drug based on metabolomics analysis. Construction method of rat model for drug screening

Method used

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  • A method for constructing a rat model for anti-oxidative aging drug screening based on metabolomics analysis
  • A method for constructing a rat model for anti-oxidative aging drug screening based on metabolomics analysis
  • A method for constructing a rat model for anti-oxidative aging drug screening based on metabolomics analysis

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Embodiment 1

[0043] [Example 1] Divide the experimental animals into normal group (blank group), model group, positive control group, high-dose group, middle-dose group and low-dose group, inject, gavage and collect urine samples

[0044] The specific method of animal treatment and sample collection is as follows: 48 SD rats were adaptively raised in an SPF environment for one week, and were randomly divided into 6 groups, namely blank control group, model group, positive control group, high-dose group, middle-dose group, etc. Dose group and low dose group, 8 rats in each group. Model group, positive control group and ellagic acid administration group were treated with 100mg·kg -1 d -1 Dosage D-galactose solution was injected subcutaneously in the neck once, on this basis, the positive control group was given 150mg·kg by intragastric administration every day -1 vitamin E; the high, medium and low dose groups were given 150, 100, 50 mg·kg-1·d by intragastric administration respectively -...

Embodiment 2

[0045] [Example 2] Prepare urine sample and obtain LC-MS spectrum data

[0046] The specific methods for the preparation of urine samples and LC-MS data acquisition are as follows: collect the urine of 6 groups of rats with metabolic cages and store them at -70°C. When testing, take urine samples and melt them on ice. min -1 , centrifuge at 4°C for 5min, take 0.5ml of the supernatant, add 0.5ml of methanol, vortex and mix well, let stand at 4°C for 3h, and then add 0.5ml of the supernatant to 3×104r·min -1 , centrifuged for 10 min, and the supernatant was taken, filtered through a 0.22 μm microporous membrane and used for LC-MS analysis. LC-MS conditions, liquid phase conditions are PHENOMEX QC MIX870 chromatographic column (50×4.6mm, 2.6μm), injection volume 1μl, column temperature 40°C, run for 30min. Mobile phase A is acetonitrile, mobile phase B is 0.1% formic acid, and the flow rate is 2ml min -1 , the elution conditions are shown in Table 1; the mass spectrometry cond...

Embodiment 3

[0047] [Example 3] Statistical methods were used to perform LC-MS data processing and pattern recognition to find out the urinary metabolomics biomarkers for the intervention of ellagic acid on D-galactose-induced aging rats

[0048] The original mass spectrum data information obtained by LC / ESI-MSn was processed by using MarkerView1.2.1 software for peak matching, peak extraction and data export. The data obtained above were imported into SIMCIAP-12.0 software for principal component analysis. Principal component analysis (PCA) for unsupervised pattern recognition, partial least squares (PLS-DA) for supervised pattern recognition and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for discriminant analysis. The differences among the sample groups of each group were compared by the score plot (Scores Plot), and the quality of the model was evaluated by parameter values ​​such as R2X, R2Y, and Q2. The closer R2X and R2Y were to 1, the more stable the ...

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Abstract

The invention discloses a constructing method of a rat model for screening drugs for resisting oxidative aging based on metabolomics analysis. The constructing method comprises the following steps: detecting and analyzing a collected urine sample by adopting an LC-MS (Liquid Chromatography-Mass Spectrometry) spectrum; carrying out clustering analysis on rat urine metabolites from six groups: a normal control group, a model group, a positive control group, an ellagic acid high dose group, a medium dose group and a low dose group by adopting PCA (Principal Component), PLS (Partial Least Square)-DA (Discriminant Analysis) and OPLS-DA (Orthogonal Partial Least Square-Discriminant Analysis) and other technologies, and screening out a potential biomarker; carrying out signal pathway analysis by applying a KEGG (Kyoto Encyclopedia of Genes and Genomes) database; annotating a metabolite molecule and analyzing related enzyme or transport protein and related properties thereof by using an HMDB (Human Metabolome Database); carrying out visual mapping on a metabolic pathway by Met PA network software; constructing the metabolic pathway of rat urine metabolomics for improving D-type galactose-induced aging by ellagic acid; and providing a metabolomics idea for screening, evaluating, researching and developing new drugs for resisting the oxidative aging.

Description

technical field [0001] The invention belongs to the technical field of metabolomics analysis, and in particular relates to a method for constructing a rat model for screening anti-oxidative aging drugs based on metabolomics analysis. Background technique [0002] Aging is a biological process affected by many complex factors such as genetics, environmental conditions, and lifestyle [1] . The theory of free radical aging holds that the oxidative damage caused by the free radicals produced by the body's cell metabolism is the key factor to accelerate the body's aging; with the aggravation of aging, various physiological functions of the body also decline, manifested as lack of energy, forgetfulness, and cold limbs. Cold, poor appetite, lack of sleep, weak waist and knees, shortness of breath and fatigue, and even facial edema, etc., are also extremely detrimental to human health and quality of life [2] . Aging is irreversible, but it can be slowed down. A large number of s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 周本宏陈鹏陈富超邱振鹏
Owner WUHAN UNIV
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