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Application of endogenous small-molecular substances in rapid detection of hepatotoxicity

A small molecular substance, endogenous technology, applied in the direction of measurement devices, material separation, analysis of materials, etc., can solve the problems of lack of specificity and sensitivity, and can not detect liver damage in time, and achieve the effect of making up for limitations

Active Publication Date: 2015-01-14
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, blood aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are two indicators commonly used to detect liver function, but they cannot detect liver injury in time due to lack of specificity and sensitivity

Method used

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  • Application of endogenous small-molecular substances in rapid detection of hepatotoxicity
  • Application of endogenous small-molecular substances in rapid detection of hepatotoxicity
  • Application of endogenous small-molecular substances in rapid detection of hepatotoxicity

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1. Reagents: Acetonitrile was purchased from Oceanpak (Gothenburg, Sweden), and formic acid was purchased from ROE (USA), both of analytical grade. Purified water was purchased from Wahaha Company (Hangzhou, China). Normal saline was purchased from Qidu Pharmaceutical Co., Ltd. (Shandong, China). Gentamicin, etimicin, isoproterenol, 5-fluorouracil, Bupleurum and carbon tetrachloride were purchased from Silan Science and Technology Co., Ltd. (Tianjin, China). Drugs were formulated with physiological saline and given to animals.

[0027] 2. Animal experiments: Animal experiments were conducted at the Institute of Biomedical Engineering, Chinese Academy of Medical Sciences (Tianjin, China). Seventy male Wistar rats (200 ± 20 g) were maintained in an SPF grade laboratory. After the rats were purchased, they were reared under controlled environmental conditions of 12 hours of day and night alternation, an ambient temperature of 25±1° C., and an ambient humidity of 50±5%. ...

Embodiment 2

[0036] Screening method for liver toxicity biomarkers based on metabolomics technology combined with SVM, including: sample pretreatment, data collection, data processing (multivariate statistical analysis), specificity investigation of biomarkers, verification and optimization of specific biomarkers and other steps. It is characterized in that: gradient elution conditions are used in data collection: 0-0.5 min, A: 99%-99%; 0.5-2 min, A: 99%-50%; 2-9 min, A: 50% -1%; 9-10 min, A: 1%-1%; 10-10.5 min, A: 1%-99%; 10.5-12 min, A: 99%-99%, where mobile phase A refers to 0.1% Formic acid in water, mobile phase B refers to 0.1% formic acid in acetonitrile; MATLAB R2010a software (USA) was used in the verification and optimization of exclusive biomarkers to establish an SVM prediction model based on liver toxicity biomarkers, the process is as follows: biomarkers The peak area in the normal saline group and each drug group is used as the input variable, and the data are randomly sele...

Embodiment 3

[0038] Practical application:

[0039] We thawed the collected serum at room temperature for detection of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum. The results showed that compared with the control group, the biochemical indicators in the Bupleurum group and the carbon tetrachloride group all had significant changes (P Figure 6 . This situation occurs because ALT is not a substance unique to the liver, it also exists in the heart tissue, so when the heart tissue is damaged, the ALT value increases. It can be seen that ALT is not specific for the detection of liver. In contrast, the specific biomarkers of hepatotoxicity that we discovered through metabolomics combined with SVM have strong sensitivity and specificity. It can be better used for the prevention, discovery and treatment of liver toxicity, make up for the limitations of existing indicators, and provide accurate detection information in a timely manner.

[0040] The specifi...

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Abstract

The invention discloses a metabonomics-based application of endogenous small-molecular substances in rapid detection of hepatotoxicity. The endogenous small-molecular substances are 12 hepatotoxicity biomarkers, i.e., L-alanine, L-phenylalanine, L-carnitine, L-carnitine palmitoyl, tryptophan, arachidonate, cholic acid, lysophosphatidylcholine (14:0), lysophosphatidylcholine (16:1), lysophosphatidylcholine (18:2), lysophosphatidylcholine (20: 3) and hemolysis phosphatidyl ethanolamine (18:2); and the endogenous small-molecular substances also refers to 3 special hepatotoxicity biomarkers, i.e. L-carnitine, cholic acid and lysophosphatidylcholine (14:0). By adopting the hepatotoxicity biomarkers, the discovery and diagnosis of the liver injury can be earlier than those of the existing biochemical detection indexes, the hepatotoxicity biomarkers can be well used for preventing, discovering and treating the hepatotoxicity, the limitation of the existing indexes can be overcome, and accurate detection information can be provided in time.

Description

technical field [0001] The present invention involves the use of metabolomics technology to find hepatotoxicity biomarkers, then investigate their specificity, and then use support vector machine (SVM) to verify and optimize them. More specifically, the application of endogenous small molecular substances in the rapid detection of liver toxicity. Background technique [0002] The liver plays an important role in the process of substance metabolism, so it becomes the main target organ damaged by toxic substances. Hepatotoxicity is a major safety concern in drug development and a common cause of preclinical trial failures and drug withdrawals. At present, blood aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are two indicators commonly used to detect liver function, but they cannot detect liver injury in time due to lack of specificity and sensitivity. Therefore, there is an urgent need for us to establish an early and accurate liver toxicity evaluation m...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 李遇伯张艳军王磊
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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