Brucea javanica oil emulsion and application thereof to inhibition of hepatitis B virus
The technology of Bruca japonica oil emulsion and hepatitis B virus, which is applied in the biological field, can solve the problems of poor treatment effect, high HBV infection rate, heavy disease and economic burden, etc., and achieves the effect of increasing practicability
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Embodiment 1
[0028] Preparation of Brucea javanica oil emulsion:
[0029] Add 0.12ml of Tween-80 and 0.125mL of glycerin to 0.5ml of javanica javanica oil (0.9g / ml), stir evenly, then gradually add sterile water to 5mL, stir evenly, filter and pack in 10ml centrifuge tubes, seal. When ready to use, sterilize by microporous membrane to get 10% Brucea javanica oil (mL / mL: 1mL medicinal solution is dissolved in 1mL sterile distilled water)) finished oil emulsion (90mg / ml), then diluted to 50mg with DMEM medium / ml mother solution, stored at 4°C for future use.
Embodiment 2
[0031] For the cytotoxicity test of duck gall oil emulsion, HepG2.215 / HepG2 / Huh7 cell suspension was used as cultured cells for comparison test. The specific steps are as follows:
[0032] First calculate the number of cells in the prepared HepG2.215 / HepG2 / Huh7 cell suspension with a cell counting plate, adjust the cell concentration to 500,000 / ml, inoculate into a 96-well culture plate, (100 μL / well, 50,000 cells / well ). Place the culture plate in an incubator (37°C, 5% CO2) for pre-culture for 24 hours. Add 10 μl of Brucea javanica oil emulsion of different concentrations to each well, each concentration has three replicate wells, and incubate the culture plate in an incubator for 24 hours. Then add 10 μL of CCK-8 solution to each well, and place the culture plate in an incubator for 1-4 hours. Measure the absorbance at 450nm with a microplate reader, and use the formula cell viability (%)=[A (dosing)-A (blank)] / [A (0 dosing)-A (blank)]×100 to calculate Brucea javanica Cy...
Embodiment 3
[0034] Inhibitory Effect of Duck Gall Oil on Hepatitis B Virus
[0035] Cell preparation:
[0036] HepG2.215 cells were inoculated into six-well cell culture plates, about 10 6 6 hours after HepG2.215 inoculation and after the cells adhered to the wall, aspirate the culture medium and add 1ml culture medium to each well, then add 40μl, 80μl and 200μl respectively (final concentrations are 1mg / ml, 2mg / ml, 5mg / ml) javanica javanica oil emulsion (50mg / ml) in the cell culture well, then replenish to 2ml every hole with medium, set up the blank control that does not add medicine simultaneously, every kind of processing does three cell duplicate wells.
[0037] HepG2 and Huh7 cells were inoculated into six-well cell culture plates, about 10 6 / well, transfect the pBSK-HBV1.3 plasmid within 16-24 hours after inoculation, and after 6 hours of transfection, suck out the medium and re-add 1ml of medium to each well, then add 40μl, 80μl, 200μl (final concentration respectively) Be 1mg...
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