Application of tubeimoside in preparing medicine for treating special subtype leukemia
A special subtype, leukemia technology, applied in the field of medicine
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Embodiment 1
[0010] Soil Fritillaria saponin C can be extracted from the soil Fritillaria bulbs by the following methods:
[0011] Crush the dried bulb of Fritillaria, sieve it, soak it with a certain amount of 5% ethanol several times, recover the ethanol under reduced pressure, and concentrate the extract until it has no alcohol smell. The fat was extracted and degreased with chloroform and n-hexane respectively, and the aqueous layer was lyophilized. The lyophilized powder was dissolved in water and separated by D101 ion exchange resin, and eluted with 30%, 70% and 100% ethanol respectively to obtain 3 fractions. Put the part eluted with 70% ethanol on a silica gel column, and elute it with a mixture of chloroform-methanol-water (7:3:1) to obtain Tubeimoside A and two other parts, one part of which is put on C 18 Reversed-phase silica gel column, eluted with 75% methanol solution to obtain the fritillary saponin B and another part; separate it by preparative HPLC, the mobile phase is 7...
Embodiment 2
[0013] CCK8 method to detect the growth inhibitory effect of fritillary saponin C on 8 kinds of leukemia cells
[0014] 2.1 Cell lines
[0015] Jurkat (acute T-cell leukemia), THP-1 (acute monocytic leukemia), L12010 (prolymphocytic leukemia), Dami (megakaryoblastic leukemia), EOL-1 (eosinophilic leukemia), RBL-1 ( Basic myeloid leukemia), MOLT-16 (adult T-cell leukemia), NCI-H929 (plasma cell leukemia). All cell lines were provided by the Affiliated Hospital of Guangdong Medical College. Cells were cultured in RPMI1640 medium containing 10% fetal bovine serum, penicillin 100U / L, streptomycin 100mg / L, 37°C, 5% CO 2 , cultured in a saturated humidity incubator, and the cells in the logarithmic growth phase were taken for experiments.
[0016] 2.2 Drugs
[0017] The imatinib solution and the fritillary saponin C solution were prepared with physiological saline, so that the final concentrations of the drugs were 1, 5, 10, 50, and 100 μmol / L.
[0018] 2.3 Test method
[0019...
Embodiment 3
[0023] Inhibition test on transplanted tumors of Dami cells in vivo Test drugs: Weigh out certain doses of fritillary saponin C and imatinib, respectively, and add normal saline to dissolve.
[0024] Administration route: intraperitoneal injection (ip).
[0025] Test animals: Dutch nude mice.
[0026] Cell line: megakaryocytic leukemia cell line (Dami), from the Affiliated Hospital of Guangdong Medical College.
[0027] Test method: 50 Dutch nude mice, weighing 18-22g. Both male and female. The Dami cell line 7-8 days after inoculation was diluted and counted at 1:10, so that the final concentration was 10 million leukemia cells / ml; 0.2ml (containing 2 million leukemia cells) was inoculated subcutaneously in the right armpit of each mouse. The next day, it was randomly divided into 5 groups: Tubeimoside C group (dose: 3mg / kg), Tubeimoside C group (dose: 8mg / kg), Tubeimoside C group (dose: 20mg / kg) blank Control group and imatinib (dose: 8mg / kg) positive control group. Tub...
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