Application of plantain herb polysaccharide in preparation of preparation for resisting porcine pseudorabies virus infection
A porcine pseudorabies and virus infection technology, applied in the field of biomedicine, to achieve the effect of rich drug sources, significant effects, and reduced cell lesions
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Embodiment 1
[0028] A preparation against porcine pseudorabies virus infection is obtained by dissolving plantain polysaccharide in 0.01mol / L PBS buffer solution, and the concentration of plantain polysaccharide is 50mg / mL.
Embodiment 2
[0030] Effects of plantain polysaccharides on cytopathic effects caused by porcine pseudorabies virus
[0031] PK-15 cells were digested with 0.25% trypsin, diluted with DMEM nutrient solution containing 5% fetal bovine serum and counted, spread into six-well plates at 5×10 5 The concentration of cells per well is appropriate, placed at 37°C, 5% CO 2 After culturing in an incubator until the cells grew to a density of 70-80% (about 17-18h), the original culture medium was discarded, and the cells were washed three times with PBS solution. / mL, 200μg / mL, 400μg / mL, 600μg / mL) after pretreatment for 1h, add PRV XJ5 strain (0.1MOI) to infect the cells, set at 37°C, 5% CO 2 One hour after infection in the incubator, wash three times with PBS solution, replace each well with DMEM nutrient solution containing 2% fetal bovine serum, infect for 24 hours and the drug has been present, observe the morphological changes and pathological changes of the cells under a microscope. according ...
Embodiment 3
[0033] The activity of plantain polysaccharides inhibiting porcine pseudorabies virus infection on PK-15 cells (Western blot, IFA and TCID 50 experiment)
[0034] (1) Western blot determination of the activity of plantain polysaccharides in inhibiting porcine pseudorabies virus infection on PK-15 cells
[0035] PK-15 cells were digested with 0.25% trypsin, diluted with DMEM nutrient solution containing 5% fetal bovine serum, and added dropwise to a 6-well plate at 5×10 5 / well concentration is appropriate, placed at 37 °C, 5% CO 2Cultivate in the incubator until the cells adhere to the wall and form a monolayer (about 17-18h), discard the original culture medium, wash the cells three times with PBS solution, suck up the residual liquid, add 1mL serum-free DMEM to each well and corresponding concentrations of plantain polysaccharides (100 μg / mL, 200 μg / mL, 400 μg / mL, 600 μg / mL), and PK-15 cells at 37 ° C, 5% CO 2 After incubating in the incubator for 1 h, infect the cells wi...
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