Application of vaccaria segetalis seed extract in promotion of cow lactation
A technology of wheat blue cabbage and extracts, which can be used in applications, medical preparations containing active ingredients, animal feed, etc., and can solve problems such as drug resistance, mutagenesis, and carcinogenicity.
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Embodiment 1
[0105] Embodiment 1, the acquisition of dairy cow mammary gland epithelial cells
[0106] 1. Obtain the mammary gland tissue of healthy mid-lactation Holstein dairy cows (raised in the Animal Husbandry High-tech Park of Northeast Agricultural University), and cultivate dairy cow mammary gland epithelial cells (methods refer to literature: Chen Jianhui. Establishment of dairy cow mammary gland epithelial cell lines [D]. Northeast Agriculture University, 2008.). The cultured mammary gland cells are all epithelioid cells, short fusiform or polygonal, uniform in shape, and closely arranged between cells (such as figure 1 shown, 200×).
[0107] 2. To identify the breast epithelial cells obtained in step 1, follow the steps below:
[0108] (1) Making cell smears: put a clean cover glass into a 6-well cell plate, and when the cells are fused to a monolayer of 80%, absorb the culture medium and wash the cells twice with PBS buffer, 5 min each time;
[0109] (2) Pre-cool methanol an...
Embodiment 2
[0119] Embodiment 2, the acquisition of barley cabbage seed extract
[0120] 1. Take dried barley seeds, crush them and pass through a 60-mesh sieve.
[0121] 2. Put the barley seed powder (1 g) obtained in step 1 into a MAS-II microwave reaction tank for microwave-assisted extraction. The extraction conditions are: solvent: 57% (volume fraction) methanol aqueous solution; temperature 65°C; liquid-solid ratio 50mL / g; microwave power 400W; extraction time 15min.
[0122] 3. Filter the extract obtained in step 2 through a 0.45 μm membrane filter to obtain a filtrate.
[0123] 4. Purify the filtrate obtained in step 3 by high performance liquid chromatography, and collect the eluate corresponding to the chromatographic peak that appeared at the 19th minute.
[0124] The detection conditions of high performance liquid chromatography are as follows:
[0125]Chromatographic column: HIQ sil C18W column ( 5μm);
[0126] Mobile phase is made up of solution A (30%) and solution B ...
Embodiment 3
[0133] Example 3, the effect of barley seed extract on the proliferation ability and vitality of dairy cow mammary gland epithelial cells
[0134] 1. Inoculate the mammary gland epithelial cells in the logarithmic growth phase prepared in Example 1 to a 12-well plate (1 × 10 in each well). 6 cells) were cultured statically at 37°C in DMEM / F12 medium (20% fetal bovine serum + 0.1% double antibody) until the cells grew to 70%-80%.
[0135] 2. After completing step 2, take the 12-well plate and perform the following grouping process:
[0136] Drug group 1: Discard the culture supernatant, add DMEM / F12 culture solution (20% fetal bovine serum + 0.1% double antibody) containing 0.1 μg / mL extract A, and let stand at 37°C for 24 hours; the addition of extract A In the form of extract mother liquor A;
[0137] Drug group 2: Discard the culture supernatant, add DMEM / F12 culture solution (20% fetal bovine serum + 0.1% double antibody) containing 1 μg / mL extract A, and let stand at 37°...
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