Application of several traditional Chinese medicine extracts in the treatment of hiv latent reactivation
A technology of extracts and traditional Chinese medicines, applied in the field of medicine, can solve problems such as poor specificity of action, toxic side effects of the body, etc.
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Embodiment 1
[0038] Preparation of plant extracts:
[0039] Purchase crude drugs such as Shegan, Huang Daphne, Liaogewang, Daphne, Tiehaitang, Sapium, Zeqi, etc. from medicinal material companies. Crush the crude drug into powder and filter through a 20-mesh sieve. Weigh the traditional Chinese medicine powder, mix it evenly with 60% ethanol-water mixture or 100% ethanol according to the drug mass:liquid volume ratio of m(g):V(ml)=1:5, soak at room temperature for more than 6 hours, preferably overnight. Then soak the traditional Chinese medicine and transfer it to the percolation tube. Every time you add a spoonful, use a glass rod to gently press it evenly, so that the medicine can be spread evenly in the tube without air bubbles. The upper end of the percolation tube is sealed with a parafilm, and 4 small holes are pierced with a needle. Control the flow rate at 3-5mL / min / kg. Always observe the flow rate during the diafiltration process, and make sure that the liquid level in the tub...
Embodiment 2
[0041] (1) HIV latent infection cell line
[0042] J-Lat cells are a common in vitro model for studying HIV-1 latent infection, which is derived from Jurkat cells infected with retroviruses expressing green fluorescent protein. When the cells were unstimulated, HIV-1 in J-Lat cells was transcriptionally silent, with no GFP expression. When treated with a latent infection reactivator, HIV-1 LTR-directed GFP expression can be activated as a marker of reactivation. The HIV latently infected cell model used in the present invention is J-Lat 6.3, J-Lat8.4 and J-Lat 11.1 cells, all of which are human T cell clones (J-Lat Full Length Clone) carrying the green fluorescent protein gene. HIV-R7 / E - The reverse transcription packaging virus of / GFP gene is prepared by infecting the human T cell line Jurkat, which contains the full-length HIV-1 gene with the deletion of env gene and nef gene, and carries the gfp gene. Under normal circumstances, this gene is not expressed, but under ce...
Embodiment 3
[0050] Activation of Chinese medicine extract composition on HIV latent cell model J-Lat 11.1
[0051] Harvest J-Lat 11.1 cells, adjust the cell concentration to 500,000 / mL, add 200 μL to each well of a 96-well plate, that is, 100,000 / well, dilute the drug, and add 1 μL of dilution to each well. Sodium butyrate was diluted 2 times, so that the final concentrations were 0mM, 1mM, 2mM, 4mM, 8mM; Sapium aconitifolia 60% alcohol extract was diluted 2 times, so that the final concentrations were 0μg / mL, 1.49μg / mL, 2.98μg / mL mL, 5.95μg / mL, 11.91μg / mL, 23.81μg / mlL; the 60% alcohol extract of Daphne genkkin was diluted 3 times, so that the final concentration was 0μg / mL, 0.206μg / mL, 0.617μg / mL, 1.85μg / mL , 5.55 μg / mL, 16.67 μg / mL. Sodium butyrate, Chinese tallow tree and yellow genkwa are combined to act on the cells according to the above concentrations. Three replicate wells were set up for each dose, and only culture medium was added to the control wells. Place the plate at 37°C...
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