Application of pectolinarin in preparation of anti-osteosarcoma medicine and anti-osteosarcoma medicine preparation
A technology of leaf glycosides and pharmaceutical preparations of chrysalis, which is applied in the field of medicine, can solve problems such as adverse reactions and large toxic and side effects of western medicines, achieve the effects of inhibiting proliferation, solving large toxic and side effects, and improving the therapeutic effect
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Embodiment 1
[0041] Example 1 Effects of Lemonfish leaf glycosides on cell proliferation
[0042] (1) MTT method was used to detect the effects of lobster leaf glycosides on the proliferation of 143B, MG-63, U2OS, Sao-2 and HOS cells.
[0043] Cells were incubated at 37°C, 5% CO 2 In the incubator, the MEM medium containing 10% fetal bovine serum was used for monolayer subculture in a 10mm culture dish, and the cells in the logarithmic growth phase were taken for experiments. Cell digestion and counting, 8000 cells per well were seeded in a 96-well plate, and adhered to the wall for 24 hours. The leaf glycosides of willowfish were dissolved in DMSO to make 50μmol·mL -1 Stock solution, diluted to 10, 20, 40, 80, 160 μM, 320 μM with MEM. Each group was added different concentrations of salivary carp leaf glycosides (10, 20, 40, 80, 160, 320 μ M), and each concentration was parallel to 6 wells. In the blank control group, an equal volume of MEM medium containing 10% fetal bovine serum was ...
Embodiment 2
[0048] Example 2 Salmonella leaf glycoside inhibits the clone formation ability of human osteosarcoma cell 143B, HOS
[0049] The 143B and HOS cells in the logarithmic growth phase were digested and counted, about 1000 cells per well were seeded in a 12-well plate, and grown adherently for 24 hours. The cells were intervened by the addition of lobster leaf glycoside at the final concentration of 50, 100, and 150 μM, and a blank control group without lobster leaf glycoside was set up. After acting for 14 days, the cells in the 12-well plate were stained with crystal violet solution, and the formation of cell colonies in each group was photographed.
[0050] figure 2 The cell proliferation results and statistical histograms of the colony formation experiment of the effect of different concentrations of salivary carp leaf glycosides on the proliferation ability of osteosarcoma cells; among them, Figure A is the result of the clone formation test of the cell proliferation, and F...
Embodiment 3
[0052] Example 3 Annexin V-FITC / PI double-staining to detect the effect of the leaf glycosides of Salix carina on the apoptosis rate
[0053] Annexin V-FITC / PI Cell Apoptosis Detection Kit (KGI, China) was used to detect the effect of different doses of fenugreek leaf glycosides intervened for 48 hours on the apoptosis rate of 143B and HOS cells. The kit includes Binding buffer, AnnexinV-FITC and Propidiμm Iodide staining solution.
[0054] 143B and HOS cells digested and counted in the logarithmic growth phase, 10 per well 6 Cells were seeded in 6-well plates and grown for 24 h. The final concentrations of 50, 100, and 150 μM lobster leaf glycosides were added to act for 48 hours, and a blank control group (control) without lobster leaf glycosides was established at the same time. Collect the supernatant and cells, centrifuge at 1000rpm for 5min, remove the medium, wash twice with PBS, and resuspend the cells in 500μL Binding buffer to make the cell concentration 2×10 6 in...
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