Application of dihydromyricetin to alleviation of intestinal cell damage caused by vomitoxin
A technology of dihydromyricetin and intestinal cells, which is applied in the field of dihydromyricetin to alleviate intestinal cell damage, and can solve problems such as the treatment of intestinal cell damage by dihydromyricetin that has not been reported
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Embodiment 1
[0043] Example 1: IPEC-J2 cell culture of porcine jejunum epithelial cells
[0044] In this example, IPEC-J2 cells of porcine jejunum epithelial cells were cultured.
[0045] Cell recovery: Take the frozen cells out of the liquid nitrogen and quickly place them in warm water at 37°C. After the frozen solution has melted, transfer them to a cell culture dish with a diameter of 10 cm and culture the cells in a medium containing 10% Serum in DMEM medium, and then placed in an incubator with 5% carbon dioxide and 37°C for cultivation.
[0046] Cell inoculation, when the cells grow to 80-90% of the culture dish area, discard the medium, wash 3 times with PBS, add 1.5mL trypsin, digest for 5 minutes, add an equal volume of complete culture medium containing 10% fetal bovine serum The base terminates digestion.
Embodiment 2
[0047] Example 2: Deoxynivalenol damages intestinal cells
[0048] In this example, DON was used to damage intestinal cells, especially porcine jejunum epithelial cells IPEC-J2 were damaged by DON.
[0049] Cell Counting Kit-8 (referred to as CCK-8) reagents can be used for simple and accurate toxicity analysis. The basic principle is: the reagent contains WST-8, chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4 -Disulfonic acid phenyl)-2H-tetrazole monosodium salt, which is dehydrogenated by the electron carrier 1-methoxy-5-methylphenazinium dimethyl sulfate (1-MethoxyPMS) The enzyme is reduced to a highly water-soluble yellow formazan product (Formazan dye). The amount of formazan produced is directly proportional to the number of living cells, and the higher the content of formazan, the greater the absorbance value, which also indicates the more living cells. This feature can therefore be used directly for cytotoxicity assays.
[0050] Pig jejunum ep...
Embodiment 3
[0054] Example 3: Dihydromyricetin can alleviate the cytotoxicity of jejunal epithelial cells
[0055] In this example, dihydromyricetin is used to alleviate the toxicity of intestinal cells. In this example, dihydromyricetin is used to alleviate the toxicity of pig intestinal cells IPEC-J2, especially dihydromyricetin is used to alleviate the toxicity of porcine intestinal cells caused by vomitoxin. Toxicity of IPEC-J2.
[0056] Cells were seeded in 96-well plates with 10,000 cells per well to allow the cells to grow adherently. In order to find the appropriate concentration of deoxynivalenol for the treated cells, the cells were divided into the following five groups for treatment: control group, 250ng / ml deoxynivalenol treatment group, 250ng / ml deoxynivalenol and 20μM dihydromyricetin co-treatment group, 250ng / ml deoxynivalenol Co-treatment group with 40 μM dihydromyricetin, 250 ng / ml deoxynivalenol and 80 μM dihydromyricetin co-treatment group.
[0057] After 24 hours ...
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