A method for constructing a model based on spleen metabolomics to study the immune mechanism of selenized amino polysaccharides on black sea bream
A technology of metabolomics and amino selenide, applied in the field of biomedical analysis, can solve the problems of poor systematicness, complicated operation and low accuracy, and achieve the effects of low cost, simple operation and high accuracy
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Embodiment 1
[0038] Embodiment 1: A method for constructing a model for enhancing the immune mechanism of black sea bream based on the study of selenized amino polysaccharides based on spleen metabolomics, the method comprising the following steps:
[0039] 1) Stop feeding the juvenile black sea bream for 1 day, and then select juvenile black sea bream with healthy body, uniform size, and initial body weight of 12.8-13.2 g and randomly divide them into the experimental group and the blank group. The blank group is fed with ordinary feed, In the experimental group, 0.6mgSe / kg selenized aminopolysaccharide was added to the common feed, and fed for 8 weeks. After the end, the juvenile black sea bream was starved for 24h, anesthetized with 60mg / L MS-222, and the spleen tissue was taken out. Each of the experimental group and the blank group selected 10 parallel samples, weighed and quickly frozen with liquid nitrogen, and then stored in an ultra-low temperature freezer at -80°C for future use; ...
Embodiment 2
[0047] Embodiment 2: The difference from Example 1 is that in step 1), the experimental group added 0.5mg Se / kg selenized amino polysaccharide to the common feed, and fed it for 10 weeks; step 2) was to get the above-mentioned 20 spare spleens Put the tissue in a centrifuge tube, add 15 times the volume of methanol frozen at -10°C, homogenize at 10000rpm for 0.5min, sonicate at 4°C for 2min, then centrifuge at 4°C and 12000rpm for 20min, take the supernatant and store it at 30°C Blow dry under nitrogen, and prepare spleen tissue samples by reconstitution with methanol aqueous solution (the volume ratio of methanol to water is 4:1); take the supernatants in the preparation of the above spleen tissue samples, mix them evenly, and dry them with nitrogen at 50°C , and redissolved with methanol aqueous solution to prepare quality control samples.
Embodiment 3
[0048] Embodiment 3: The difference from Example 1 is that in step 1), the experimental group added 0.7mg Se / kg selenized amino polysaccharide to the common feed, and fed it for 9 weeks; step 2) was to get the above-mentioned 20 spare spleens Put the tissue in a centrifuge tube, add 5 times the volume of methanol frozen at -30°C, homogenize at 14000rpm for 0.5min, sonicate at 4°C for 5min, then centrifuge at 4°C and 14000rpm for 10min, take the supernatant at 50°C Blow dry under nitrogen, and prepare spleen tissue samples by reconstitution with methanol aqueous solution (the volume ratio of methanol to water is 4:1); take the supernatants in the preparation of the above spleen tissue samples, mix them evenly, and dry them with nitrogen at 30°C , and redissolved with methanol aqueous solution to prepare quality control samples.
[0049] Before preprocessing the analytical data, the quality and validity of the obtained analytical data are verified first. The total ion current c...
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