Application of triptonide in preparation of medicine for treating acute monocyte leukemia
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A kind of technology of triptolide ketone and acute single, applied in the field of leukemia drugs
Pending Publication Date: 2020-07-14
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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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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Abstract
The invention provides application of triptonide in preparation of a medicine for treating acute monocyte leukemia, and belongs to the technical field of leukemia medicines. According to the application of the triptonide in preparation of the medicine for treating the acute monocyte leukemia, acute monocyte leukemia cells U937 are taken as research objects, U937 cells are treated by a triptonide solution, the cell activity of the U937 cells is detected by an MTT method, and the result shows that the triptonide has an effect of inhibiting proliferation of the U937 leukemia cells. The influenceof the triptonide on apoptosis of the U937 cells is detected by an Annexin V/PI double staining method, and the result shows that the triptonide can induce apoptosis of the U937 cells, and the apoptosis is more obvious along with increase of concentration. Therefore, the triptonide can inhibit proliferation of the acute leukemia cells and induce apoptosis, so that treatment of the acute monocyte leukemia is realized.
Description
technical field [0001] The invention belongs to the technical field of leukemia drugs, and in particular relates to the application of triptolide in the preparation of drugs for treating acute monocytic leukemia. Background technique [0002] Leukemia, also known as blood cancer, is a malignant proliferative disease of hematopoietic cells. Leukemia cells proliferate and accumulate in the bone marrow, lymph nodes and other hematopoietic systems due to their characteristics such as uncontrolled proliferation, blocked differentiation, and apoptosis, and infiltrate other tissues and organs in the body. organs, causing the normal hematopoietic function to be inhibited, resulting in anemia, hepatosplenomegaly and other symptoms. [0003] According to the pathogenesis of leukemia, it is divided into acute myelocytic leukemia (AML), chronic myelocytic leukemia (CML), acute lymphocytic leukemia (ALL) and chronic lymphocytic leukemia (CML). Leukemia, CLL). [0004] Acute myelogenous...
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IPC IPC(8): A61K31/585A61P35/02A61K36/37
CPCA61K31/585A61K36/37A61P35/02
Inventor 谭余庆刘丽李慧杰
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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