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Model construction method for researching black sea bream immune mechanism enhancement by selenized glycosaminoglycan based on liver metabonomics

A technology of metabolomics and amino selenide, applied in the field of biomedical analysis, can solve problems such as low accuracy, high cost, and complicated operation

Active Publication Date: 2019-10-18
舟山出入境检验检疫局综合技术服务中心
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Problems solved by technology

[0005] The present invention aims at overcoming the technical problems such as poor system, low accuracy, high cost and complicated operation in the current evaluation method for studying the immune mechanism of selenized amino polysaccharides on black sea bream, and proposes a method based on liver metabolomics to study selenium Construction method of a model for enhancing the immune mechanism of black sea bream with aminopolysaccharides

Method used

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  • Model construction method for researching black sea bream immune mechanism enhancement by selenized glycosaminoglycan based on liver metabonomics
  • Model construction method for researching black sea bream immune mechanism enhancement by selenized glycosaminoglycan based on liver metabonomics
  • Model construction method for researching black sea bream immune mechanism enhancement by selenized glycosaminoglycan based on liver metabonomics

Examples

Experimental program
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Embodiment 1

[0040] A method for constructing a model for enhancing the immune mechanism of black sea bream based on the study of selenized amino polysaccharide based on liver metabolomics, the method comprising the following steps:

[0041] 1) Stop feeding the black sea bream juveniles for 1 day, and then select the black sea bream juveniles with healthy physique, uniform size, and an initial weight of 12.8~13.2g into the experimental group and the blank group. The blank group is fed with ordinary feed. The experimental group was fed with 0.6mg Se / kg selenized amino polysaccharides in ordinary feed and fed for 8 weeks. After the end, the juvenile black sea bream was starved for 24 hours and anesthetized with 60 mg / L MS-222. The liver tissue was taken out. Among them, the experimental group and the blank group each select 10 parallel samples, weigh them and quickly freeze them with liquid nitrogen, and then store them in an ultra-low temperature refrigerator at -80°C for later use;

[0042] 2) ...

Embodiment 2

[0050] The difference from Example 1 is that in step 1), the experimental group added 0.5 mg Se / kg selenized amino polysaccharide to ordinary feed and fed for 10 weeks; step 2) was to take the above-mentioned spare 20 liver tissues in a centrifuge tube , Add 15 times volume of methanol frozen at -10℃, homogenize at 10000rpm for 0.5min, centrifuge at 4℃, 12000rpm for 5min, take the supernatant and blow dry at 30℃ with nitrogen, and use methanol aqueous solution (methanol and water The volume ratio is 4:1) The liver tissue samples are prepared by reconstitution, and the liver tissue sample preparation is completed within 12 minutes; take the supernatants from the above liver tissue sample preparations and mix them evenly, blow dry at 50 ℃ with nitrogen, and use methanol The aqueous solution is reconstituted to prepare a quality control sample.

Embodiment 3

[0052] The difference from Example 1 is that in step 1), the experimental group added 0.7 mg Se / kg selenized amino polysaccharide to ordinary feed and fed for 9 weeks; step 2) was to take the above-mentioned spare 20 liver tissues in a centrifuge tube , Add 5 times the volume of methanol frozen at -30℃, homogenize at 14000rpm for 0.5min, centrifuge at 4℃, 14000rpm for 3min, take the supernatant and blow dry at 50℃ with nitrogen, and use methanol aqueous solution (methanol and water The volume ratio is 4:1) The liver tissue samples are prepared by reconstitution, and the liver tissue sample preparation is completed within 12 minutes; take the supernatants from the above liver tissue sample preparations and mix them evenly, blow dry at 30°C with nitrogen, and use methanol The aqueous solution is reconstituted to prepare a quality control sample.

[0053] Before preprocessing the analysis data, first verify the quality and validity of the analysis data obtained. The total ion curre...

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Abstract

A model construction method for researching black sea bream immune mechanism enhancement by selenized glycosaminoglycan based on liver metabonomics relates to the field of biomedical analysis, and comprises the following steps: 1) uniformly dividing young black sea bream into an experimental group and a blank group; 2) adding selenized glycosaminoglycan into the feed of the experimental group; 3)carrying out hunger treatment on the young black sea bream in the experimental group and the blank group, anesthetizing, taking out liver tissues, and storing for later use; 4) preparing a liver tissue sample, preparing a quality control sample in the same time, and using ultra-high performance liquid chromatography-time-of-flight mass spectrometry technology for treatment; and (5) analyzing and processing the collected liver metabonomics data of the black sea breams, identifying and screening out liver metabonomics biomarkers for enhancing the immune mechanism of the black sea breams relatedto the selenide glycosaminoglycan, and constructing and analyzing metabolic pathways pointed by the liver metabonomics biomarkers. The construction method is good in systematicness, high in accuracy,low in cost and simple to operate.

Description

Technical field [0001] The invention relates to the field of biomedicine analysis, in particular to a method for constructing a model for enhancing the immune mechanism of black sea bream by studying selenized amino polysaccharide based on liver metabolomics. Background technique [0002] As an essential trace element in the body, selenium plays an important role in the growth of the body and the maintenance of body health through the forms of selenoprotein and selenium-dependent enzyme cofactors. At present, there are more researches on organic selenium, such as selenized amino polysaccharides, which have relatively low biological toxicity and are easy to be absorbed and utilized. Selenized amino polysaccharides are obtained by combining polysaccharides with inorganic selenium through chemical modification methods. The physiological and pharmacological functions of selenium and polysaccharides are optimized. The biological activities are generally higher than polysaccharides and...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N30/06G01N30/14G01N30/34G01N30/86G01N30/88
CPCG01N30/02G01N30/72G01N30/06G01N30/14G01N30/34G01N30/8686G01N30/88G01N2030/146G01N2030/062G01N2030/8813G01N2030/8818G01N2030/8836G01N2030/8831G01N2570/00G01N2560/00
Inventor 周秀锦张静邵宏宏晁铎源宋立玲
Owner 舟山出入境检验检疫局综合技术服务中心
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