Traditional Chinese medicine extract for inducing apoptosis of tumor cells and preparation process and application thereof
A technology of tumor cell apoptosis and extraction, which is applied in the field of extraction of Chinese herbal medicines, can solve the problems of tumor cell apoptosis and achieve the effects of inducing tumor cell apoptosis, inhibiting tumor cell proliferation, and low toxic and side effects
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Embodiment 1
[0015] Example 1 Preparation of dry powder of water extract of Phyllanthus sibiricum
[0016] 1kg of dry product of Caulis edulis, crushed, placed in a reflux extraction device, and extracted twice with a concentration of 95% (v / v) ethanol 12L and 70% (v / v) ethanol 10L each with reflux for 2 hours each time, discarded Remove the alcohol extract, add 15L of water to the obtained medicine residue, decoc 2 times, 2h each time, combine the two water extracts, filter, concentrate into a thick paste, freeze-dry, pulverize into fine powder, pass a 100-mesh sieve, and obtain wool The dry powder of the water extract of leaf white powder vine.
Embodiment 2
[0017] Example 2 Experimental study on the inhibition of the growth of tumor cells by the water extract of Phyllanthus sibiricum
[0018] Take the dry powder of the water extract of C. chinensis prepared in Example 1, and dissolve the dry powder in 1640 complete culture solution containing 10% fetal bovine serum to make the concentration of the water extract respectively 150, 200, 250, 300, 400, 500 and 600μg / mL are reserved. Human laryngeal cancer Hep2 cell line, using 1640 complete culture medium containing 10% fetal bovine serum at 37℃ constant temperature, 5% CO 2 , Cultivate under 95% saturated humidity, take Hep in logarithmic growth phase 2 Cells at 1×10 per well 4 Each was seeded in a 96-well plate with 200μL per well. After overnight incubation, the supernatant was discarded, and the prepared aqueous extract solutions of Caulis chinensis with different concentrations (150, 200, 250, 300, 400, 500, 600 μg / mL) were added to the culture plate, and a blank control group ( O...
Embodiment 3
[0020] Example 3 Experimental study of Aqueous extract of Phyllanthus sibiricum inducing tumor cell apoptosis
[0021] Human laryngeal cancer Hep2 cell line, using 1640 complete culture medium containing 10% fetal bovine serum at 37℃ constant temperature, 5% CO 2 , Culture under 95% saturated humidity conditions. Take Hep in logarithmic growth period 2 Cells to 2×10 5 / Well was inoculated into a 6-well plate covered with a cover glass, using different concentrations (150, 250, 300μg / mL) of the aqueous extract of Caulis serrata (prepared in Example 1) and 0.5μg / mL doxorubicin respectively Treated for 48h, and set up a negative control group. After incubation, the supernatant was discarded, the cell slide was washed three times with PBS, incubated in 10μg / mL Hoechst 33258 staining solution for 10 min in the dark, the cell slide was taken out and washed with PBS three times, and observed under a fluorescence microscope.
[0022] After Hoechst 33258 fluorescent staining, the cells in ...
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