Application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis

A mammary epithelial cell and astragalus polysaccharide technology is applied in the application field of astragalus polysaccharide in antagonizing the apoptosis of dairy cow mammary epithelial cells, which can solve the problems of drug residues, endanger human health, cannot be completely cured, etc., achieve feasible results, and test material safety. Effect

Inactive Publication Date: 2015-07-08
ANHUI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These methods have their own strengths, although they cannot be cured completely, they can all play a certain role.
However, the side effects of these methods cannot be ignored, especially drug residues, which indirectly endanger human health.

Method used

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  • Application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis
  • Application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis
  • Application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis

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Embodiment 1

[0041] An application method of astragalus polysaccharide in antagonizing cow mammary gland epithelial cell apoptosis, comprising the following steps:

[0042] (1) Cultivate dairy cow mammary epithelial cells, and obtain 2-3 generations of cells in logarithmic growth phase for subsequent experiments;

[0043] (2) Grouping and processing of cells:

[0044] ① Take the 2-3 generation logarithmic growth phase dairy cow mammary epithelial cells to prepare 1×10 4 cells / mL of cell suspension, inoculated in 96-well cell culture plate at 150 μL per well, at 37°C, CO 2 After conventional culture in an incubator with a volume fraction of 5% for 24 hours, they were randomly divided into the following 5 test groups: (i) blank control group; (ii) lipopolysaccharide (Lipopolysaccharide, LPS) model group; (iii) the first astragalus polysaccharide Model group; (iv) the second astragalus polysaccharide model group; (v) the third astragalus polysaccharide model group; each group was repeated o...

Embodiment 2

[0053] An application method of astragalus polysaccharide in antagonizing cow mammary gland epithelial cell apoptosis, comprising the following steps:

[0054] (1) Obtain dairy cow mammary gland epithelial cells by tissue block culture method, use DMEM / F12 culture medium containing 10% volume fraction of calf serum as the basal medium, at 37°C, CO 2 Cultured in an incubator with a volume fraction of 5%, after purification, 2-3 passages of logarithmic growth phase cells were used for subsequent experiments.

[0055] (2) Grouping and processing of cells:

[0056] ① Take the 2-3 generation logarithmic growth phase dairy cow mammary epithelial cells to prepare 1×10 4 cells / mL of cell suspension, inoculated in 96-well cell culture plate at 150 μL per well, at 37°C, CO 2 After conventional culture in an incubator with a volume fraction of 5% for 24 hours, they were randomly divided into the following 5 test groups: (i) blank control group; (ii) lipopolysaccharide (Lipopolysacchari...

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Abstract

The invention discloses an application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis. The method comprises the following steps: (1) obtaining 2-3 generations of dairy cow mammary epithelial cells; (2) grouping and processing cells; (3) detecting cell survival rate employing a CCK-8 method; (4) detecting a cell cycle employing flow cytometry; (5) detecting the change condition of nucleic acid of apoptotic cells employing a DNA ladder; (6) observing the internal morphological structure change of the cells through a transmission electron microscope; (7) detecting inflammation-associated cytokines content in a cell supernatant employing radioimmunoassay; and (8) detecting inflammation-related enzyme activity in the supernatant employing a biochemical kit method. Through the method disclosed by the invention, the action of the astragalus polysaccharide for antagonism of dairy cow mammary epithelial cells apoptosis is discussed; and the method is efficient, simple, convenient, feasible, and accurate in result.

Description

technical field [0001] The invention relates to an application method of astragalus polysaccharide; in particular, it relates to an application method of astragalus polysaccharide in antagonizing cow mammary gland epithelial cell apoptosis. Background technique [0002] Cow mastitis seriously affects the milk production of dairy cows, reduces milk quality, endangers the health of consumers, prolongs the postpartum estrus period of dairy cows, increases the elimination rate of dairy cows, and reduces the economic benefits of dairy farming. At present, there are many treatment methods for dairy cow mastitis, such as chemical drug therapy, genetic engineering vaccine therapy, cytokine therapy, bacteriocin therapy, hormone therapy, and anti-mastitis pathogenic bacteria compound egg yolk antibody therapy. These methods have their own strengths, although they cannot be completely cured, they can all play a certain role. However, the side effects of these methods cannot be ignored...

Claims

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Application Information

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IPC IPC(8): C12Q1/02
Inventor 丁月云殷宗俊张晓东孟云张威付坤张梦琦黄龙王源朗朱树娇
Owner ANHUI AGRICULTURAL UNIVERSITY
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