Application of platycodin D in preparing medicaments for inhibiting angiogenesis
A technology of platycodon saponin and angiogenesis, which is applied in the field of medicine and can solve the problems that platycodon saponin D has not been reported.
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Embodiment 1A
[0031] Example 1A Platycodon saponin D inhibits the proliferation of human umbilical vein endothelial cells (HUVEC)
[0032] The proliferation of HUVEC was detected by CCK-8 method. CCK-8 reagent contains WST–8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene) -2H-tetrazolium monosodium salt), which is reduced by the dehydrogenase in the mitochondria of the cell under the action of the electron carrier 1-Methoxy PMS (1-methoxy-5-methylphenazine dimethyl sulfate) to have Highly water-soluble yellow formazan (Formazan) product. The amount of formazan produced is directly proportional to the number of viable cells. The light absorption value was measured at a wavelength of 450 nm by an enzyme-linked immunosorbent detector, which can indirectly reflect the number of living cells. The effect of drugs on the proliferation of HUVCE can be evaluated by this method.
[0033] Methods: HUVEC cells were seeded in 96-well plates and cultured. After the c...
Embodiment 1B
[0037] Example 1B Platycodon saponin D inhibits scratch migration of human umbilical vein endothelial cells (HUVEC)
[0038]Methods: HUVEC cells were seeded in 12-well plates and cultured. After the cell growth reached 90% confluence, platycodon saponin D was added to the wells at the following concentrations of 0.3, 1, 3, 10, and 30 μM, and after 24 h of action, fresh complete medium without drugs was added to each well, and placed at 37 °C The incubator continued to incubate for 48 h, and the EVOS inverted microscope was used to take pictures. The wells without platycodon saponin D were used as the control.
[0039] Calculate the inhibition rate according to the following formula:
[0040] Inhibition rate (%) = (end point width sample - end width control ) / initial scratch width × 100%
[0041] RESULTS AND CONCLUSIONS: Please refer to the attached figure 1 B, with figure 1 B is a photograph of platycodon saponin D inhibiting the proliferation of HUVEC. As shown in ...
Embodiment 1C
[0043] Example 1C Platycodon saponin D inhibits the horizontal movement of human umbilical vein endothelial cells (HUVEC)
[0044] Using Cell Motility Kit (Cell Motility Kit) to quantitatively calculate cell motility, this experiment can predict the therapeutic effect of potential drugs in certain fields. In this experiment, cells are seeded in a 96-well plate covered with extremely small fluorescent particles. When the cells move, they will phagocytize or push away the fluorescent particles on the surface, leaving a motion estimation area without fluorescence. The area of this area is the same as that of the cells. Athletic ability was positively correlated. The area of motion area can be calculated by high-content screening analyzer and relevant statistical software.
[0045] Methods: Type Ⅰ collagen was used to coat the 96-well plate, the solution containing fluorescent particles was added, incubated at 37 ℃ in the dark for 1 h, and washed 5 times with washing soluti...
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