Novel application of dictamnine
A technology of leuconine and mast cells, applied in the field of medicine, can solve problems such as ineffective effects, high costs, and long treatment courses
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Embodiment 1
[0018] The purpose and method of the experiment: No matter what kind of drug, if the concentration is too high, it will produce cytotoxicity to the cells. If the concentration of whitewash is too high, it will have certain toxicity to P815 cells. In order to detect the toxicity of whitewash to P815 cells, CCK-8 experiment was used to detect the cell survival rate of P815 cells stimulated with different concentrations of whitewash for 24 hours, so as to determine the use concentration of whitewash that has no growth inhibitory activity on the cells. The specific steps are as follows:
[0019] (1) Culture and passage of mast cell line P815 cells
[0020] A. Cultivation of P815 cells: P815 was cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin and streptomycin at 37°C and 5% CO 2 Environment;
[0021] B. Subculture of P815 cells: When the cell density in step A reaches 70%, it needs to be subcultured. Collect the cell culture solution in the...
Embodiment 2
[0029] The purpose and method of the experiment: In order to verify whether whitewashine can inhibit mast cell degranulation, mast cells were pretreated with different concentrations of whitewash, and whether it could inhibit the release of mast cell degranulation marker β-hexosaminidase was tested. Specific steps are as follows:
[0030] (1) Culture and passage of mast cell line P815 cells
[0031] A. Culture of P815 cells: P815 cultured in high-glucose DMEM medium containing 10% fetal bovine serum (FBS), 1% penicillin and streptomycin, cultured at 37°C, 5% CO 2 Environment;
[0032] B. Subculture of P815 cells: When the cell density in step A reaches 80%, it needs to be subcultured. Collect the cell culture solution in the culture bottle in a centrifuge tube, rinse with preheated phosphate buffered saline solution three times, add an appropriate amount of trypsin, After the cells are completely digested, add the cell culture medium to neutralize the trypsin, pipette evenly...
Embodiment 3
[0040] The purpose and method of the experiment: To detect the effect of leuconine on the expression of IL-4, IL-6, IL-12 and IL-13 at the mRNA level after P815 activation, the specific steps are as follows:
[0041] (1) Culture and passage of mast cell line P815 cells
[0042] A. Cultivation of P815 cells: P815 was cultured in high-glucose DMEM medium containing 10% fetal bovine serum and 1% penicillin and streptomycin at 37°C and 5% CO 2 Environment;
[0043] B. Subculture of P815 cells: When the cell density in step A reaches 70%, it needs to be subcultured. Collect the cell culture solution in the culture bottle in a centrifuge tube, rinse with preheated phosphate buffered saline solution three times, add an appropriate amount of trypsin, After the cells are completely digested, add the cell culture medium to neutralize the trypsin, pipette evenly, centrifuge at 800rpm for 3 minutes, remove the supernatant, add fresh high-sugar DMEM medium, pipette and mix the cells, and ...
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