Application of triptonide in preparation of medicine for treating acute monocyte leukemia
A kind of technology of triptolide ketone and acute single, applied in the field of leukemia drugs
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Embodiment 1
[0033] Detection of cell viability of triptolide against acute monocytic leukemia cell line U937
[0034] experimental method:
[0035] MTT method was used to determine the inhibitory effect of different concentrations of triptolide on the proliferation of U937 leukemia cells.
[0036] Cultivate U937 cells to the logarithmic growth phase. Experiments are carried out when the cells are in good condition. 5000 cells / well are seeded in a 96-well plate, and 100 mmol / L of prepared RPMI1640 medium containing 5% FBS is diluted. Triptolide was diluted to 640nmol / L, 2ml, and diluted to 320nmol / L in a 96-well plate, 160nmol / L, 80nmol / L, 40nmol / L, 20nmol / L, 10nmol / L, 5nmol / L, 2.5 nmol / L, set up 6 replicate wells for each concentration, set up RPMI1640 medium negative control with 5% FBS, fill with sterile PBS for four weeks, and 5% CO 2 , After incubation at 37°C for 48 and 72 hours, add 20 μl of 5% MTT solution to each well, continue to incubate for 4 hours, add 100 μl of 10% SDS cont...
Embodiment 2
[0042] Experimental method for detecting the effect of triptolide on the apoptosis of acute monocytic leukemia cells U937:
[0043] Cultivate U937 cells to the logarithmic growth phase, and carry out the experiment when the cells are in good condition, inoculate 10,000 cells / well in a 6-well plate, dilute the prepared 100mM Tripterygium wilfordii in RPMI1640 medium containing 5% FBS Esterone was diluted to 40nmol / L, 20nmol / L, 10nmol / L, 5nmol / L, 2.5nmol / L, and each concentration was set up 3 duplicate wells, with the RPMI1640 medium of 5%FBS as contrast (control group), in 5%CO 2 After incubation at 37°C for 72 hours, the cells were collected, centrifuged at 200 g to discard the supernatant, washed twice with sterile PBS, added 5 μl of FITCAnnexin to incubate the treated cells for 10 minutes, then added 5 μl of PI, and placed in a room temperature environment for 5 minutes in the dark. Add 100 μl 1×BindingBuffer and mix thoroughly, and detect by flow cytometry within 1 hour. ...
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