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Test method for characterizing smoke exposure biological effect based on metabonomics

A metabolomics and test method technology, which is applied in the field of tests based on metabolomics to characterize the biological effects of smoke exposure, and achieves the effect of simple operation.

Inactive Publication Date: 2017-06-09
CHINA TOBACCO JIANGXI IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the use of metabolomics to characterize the biological effects of smoke exposure.

Method used

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  • Test method for characterizing smoke exposure biological effect based on metabonomics
  • Test method for characterizing smoke exposure biological effect based on metabonomics
  • Test method for characterizing smoke exposure biological effect based on metabonomics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1) After one week of adaptive feeding, the rats were immediately divided into: blank group, control group and experimental group; the cigarettes were fixed in the terrarium by a metal sleeve, and each rat smoked 1 cigarette per day. The rats were fasted for 12 hours before and after the operation, and they had free drinking water.

[0030] 2) Animal treatment and sample collection: After 7 days of exposure to smoke, the rats were weighed. After anesthesia, about 6-8 mL of blood was taken from the hepatic portal vein and placed in a 10 mL centrifuge tube treated with heparin sodium. Centrifuge at 3000 rpm for 10 min, take the upper layer of plasma, and store each biological sample at -80°C.

[0031] 3) Before LC-MS analysis, take 100μL of the frozen-thawed plasma sample, add 400μL of methanol, vortex for 1min, mix well to precipitate the protein, then centrifuge at 13000rpm at 4℃ for 15min, take the supernatant and add 300μL of ultrapure Dilute with water and filter with 0.2...

Embodiment 2

[0038] 1) After one week of adaptive feeding, the rats were immediately divided into: blank group, control group and experimental group; the cigarettes were fixed in the terrarium by a metal sleeve, and each rat smoked 1 cigarette a day. The rats were fasted for 12 hours before and after the operation, and they had free drinking water.

[0039] 2) Animal handling and sample collection: After exposure to smoke for 7, 14, and 30 days, the rats are weighed, and the rats’ urine for 24 hours is collected in a metabolic cage. After anesthesia, about 6-8 mL of blood is taken from the portal vein of the liver. , Put it into a 10mL centrifuge test tube treated with heparin sodium, quickly centrifuge at 3000rpm for 10min, take the upper layer of plasma, and store each biological sample at -80°C.

[0040] 3) Before LC-MS analysis, take 100μL of frozen-thawed plasma or urine sample, add 400μL methanol, vortex for 1min, mix well to precipitate the protein, then centrifuge at 13000rpm at 4℃ for ...

Embodiment 3

[0047] 1) After one week of adaptive feeding, the rats are then divided into: blank group, control group and experimental group; the cigarettes are fixed in the terrarium by a metal sleeve, and each rat is exposed for 20 minutes a day to control the smoke shading rate of cigarettes The control temperature is 22±2°C, the humidity is maintained at 21±0.5%, the oxygen concentration is maintained at 21±0.5%, and the pressure is 101325±40Pa. The rats were fasted for 12 hours before and after the operation, and they had free drinking water.

[0048] 2) Animal handling and sample collection: Weigh the rats at 7 days, 14 days and 30 days after smoke exposure. After anesthesia, take the rat lung tissue, wash it with normal saline and absorb the water with filter paper, then weigh it , Store at -80℃.

[0049] 3) Pretreatment of lung tissue samples: Take the lung tissue samples after freezing and thawing, and homogenize them by adding physiological saline at 1:3 (g / mL). Take 200μL of homoge...

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Abstract

The invention discloses a test method for characterizing the biological effects of smoke exposure based on metabolomics. The test method adopts the metabolomics analysis method, and the biomarker groups related to the biological effects of smoke exposure are tested in different drug administration groups. The changes between groups are used to characterize the biological responses of experimental animals placed in the smoke exposure environment. The present invention is easy to operate, and evaluates the influence of mainstream smoke on endogenous metabolites in rats, and compares the similarities and differences between the effects of natural herbal added tobacco and ordinary cigarettes on endogenous metabolites, in order to evaluate the harm reduction effect of cigarettes and its associated mechanisms provide a reliable tool.

Description

Technical field [0001] The invention belongs to the technical field of research on the biological effects of smoke exposure, and in particular relates to a test method for characterizing the biological effects of smoke exposure based on metabolomics. Background technique [0002] Tobacco is the whole plant of Nicotianatabacum L., a solanaceous plant. Polycyclic aromatic hydrocarbons (PAHs) and free radicals formed during the combustion process of cigarettes. Among them, polycyclic aromatic hydrocarbons are the most abundant and also the most important carcinogen in smoke; on the one hand, free radicals can be converted into polycyclic aromatic hydrocarbons under high temperature conditions. On the other hand, free radicals are also an important cause of various diseases caused by oxidative damage in the body, and even cancer. As smoking and health problems become increasingly prominent, research on tobacco harm reduction technologies has become a hot spot in the international to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/00G01N30/02G01N30/06G01N24/08
CPCG01N33/0004G01N24/08G01N30/02G01N30/06
Inventor 蔡继宝罗国安苏加坤谢媛媛郭磊王义明罗娟敏
Owner CHINA TOBACCO JIANGXI IND CO LTD
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