Application of ranunculus japonicus extract in preparing anti-tumor and anti-tumor angiogenesis drugs
A technique for angiogenesis and extraction, which is used in anti-tumor drugs, applications, drug combinations, etc.
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Embodiment 1
[0043] Example 1: Inhibitory effect of buttercup extract on tumor angiogenesis in vitro
[0044]Mouse liposarcoma cells SW872 were placed in DMEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Incubator cultivation. Take the cells in the logarithmic growth phase, and use 4*10 4 / mL cells were planted in a 24-well plate pre-coated with a layer of fibrin, placed at 37 ° C, 5% CO 2 Incubator for 24 hours. After 24 hours, different concentrations (1-1000ug / mL) of ranunculus extract were added, with the blank culture solution as the control and 5uM 5-Fu as the positive control, after 36 hours of cultivation, the supernatant was carefully removed, and stained with Wright-Gimsa solution. After staining, select a typical area of each well to take pictures. The above experiments were repeated 3 times.
[0045] The result is as Figures 1 to 17 As shown, various buttercup extracts have very good inhibitory effects on angiogenesis of tumor cells in vitro.
Embodiment 2
[0046] Example 2: Inhibitory effect of buttercup extract on the proliferation of various tumor cells
[0047] Place tumor cell lines 2C4G2, U937, 2C4, Dami, Namalva, K562, PMKO102-3#, BAEC, HMY2-CIR in DMEM or RPMI 1640 culture medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Incubator cultivation. The cells in the logarithmic growth phase were taken and seeded in a 96-well plate at an appropriate concentration, wherein the drug was administered while the suspended cells were seeded on the plate, and the drug was administered to the adherent cells 24 hours later. Put at 37℃, 5% CO 2 Incubator for 24 hours. After 24 hours, different concentrations (1-400ug / mL) of ranunculus extract were added to the adherent cells. The blank culture solution with cells was used as the control, and the blank culture solution without cells was used as the blank control. After 46 hours of culture, Alamar blue reagent was added. 10uL / well, placed in a 37°C, 5% CO2 incubator for 2 hours ...
Embodiment 3
[0054] Example 3: Inhibitory effect of buttercup extract on Matrigel tumor angiogenesis in vitro
[0055] Take 150uL of melted BD Matrigel, add it to a 48-well plate, tap it to make it evenly distributed, and incubate at 37°C for 1 hour to make Matrigel polymerize into a gel. Take the tumor cells SW872 and 3B11 in the logarithmic growth phase, and use 4*10 5 / mL cells were mixed with equal volumes of RPMI 1640 culture medium containing different concentrations of ranunculus extract, seeded on Matrigel at 200uL / well, and incubated at 37°C, 5% CO 2 After culturing in the incubator for several hours, stain with Wright-Gimsa staining solution, observe and take pictures under a microscope.
[0056] Such as Figure 18 and Figure 19 As shown, the results showed that the G component of buttercup extract had a good inhibitory effect on the in vitro Matrigel model of two kinds of tumor cells, and it was dose-dependent. In the control group, the blood vessels were thick, and the tub...
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