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Application of alantolactone serving as anti-angiogenesis drug

A technology of inulin and angiogenesis, which is applied in the field of medicine, can solve the problems that inulin has no anti-angiogenesis effect, and achieve the effect of treating breast tumors

Inactive Publication Date: 2017-10-20
SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no report about the anti-angiogenic effect of inulin

Method used

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  • Application of alantolactone serving as anti-angiogenesis drug
  • Application of alantolactone serving as anti-angiogenesis drug
  • Application of alantolactone serving as anti-angiogenesis drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1C

[0027] Embodiment 1CCK-8 method detects HUVEC cell viability

[0028] HUVEC cells at 2.5×10 4 Cell density per mL was seeded in a 96-well plate (200 uL per well). After culturing for 24 hours until the cells reached 80% confluency, the culture medium was replaced with inulin solutions with concentrations of 1, 3, 10, 30, and 100 μM. After acting for 48 hours, suck out the drug-containing medium, add CCK-8 diluted 1:10 with the medium to each well, and place it in an incubator to continue incubating for 2 hours, then use a microplate reader to measure CCK-8 in the plate at 450nm OD value of the solution. At the same time, the wells containing CCK-8 solution and medium but without cells were set as blank controls, and the wells containing cells but without drugs were set as controls. The formula for calculating cell viability is: cell viability (%)=[(ODsample–ODblank) / (ODcontrol–ODblank)]×100%.

Embodiment 2

[0029] Example 2 Cell Motility Inhibition Experiment

[0030] Cellomics Cell Motility Kit was used to measure the movement track of migrating cells to investigate the motility of HUVEC. Add blue fluorescent microsphere powder to the 96-well plate coated with collagen-I (collagen-I), and incubate for 1 h in the dark. The incubated 96-well plate was soaked 5 times with PBS. Digest and centrifuge HUVEC and resuspend, adjust the cell density to 5×10 3 / mL. The final concentrations of inulin were diluted with cell suspension to 1, 3, 10, 30 and 100 μM respectively, and 100 μL of drug-containing cell suspension was added to a 96-well plate covered with blue microsphere powder. At the same time, the no-drug group was set as a control. Then, the culture plate was placed in an incubator to continue culturing for 20 h, and then 200 μL of preheated 5.5% fixation solution was directly added to each well, and allowed to act for 1 h at room temperature. The 96-well plate was washed 3 ti...

Embodiment 3

[0031] Example 3 Cell Migration Inhibition Experiment

[0032] Add 600 μL of culture solution containing 20 ng / mL VEGF165 to the lower chamber of a Corning Transwell 24-well plate. Digest centrifuged HUVEC cells with a cell concentration of 2 x 10 4 / mL of cell suspension diluted inulin to a final concentration of 1, 3, 10, 30 and 100 μM, respectively, and 100 μL of drug-containing cell suspension was added to the upper chamber. At the same time, the no-drug group was set as the control group. Place the culture plate in the cell incubator, continue to culture for 8 hours, suck out the culture solution in the well, add 600 μL of 4% paraformaldehyde to fix the migrated HUVEC cells for 15 minutes, gently wipe off the non-migrated cells on the surface of the upper chamber with a cotton swab, and then Add 300μL of 0.1% crystal violet solution to the lower chamber for 25min staining. Finally, wash five times with PBS. Cell migration was observed and photographed under a Zeiss in...

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Abstract

The invention relates to new application of alantolactone, in particular to application of the alantolactone serving as an anti-angiogenesis drug. The application has the advantages that the activity of an HUVEC (Human Umbilical Vein Endothelial Cell) is detected through a CCK-8 method, and a cell mobility inhibition experiment, a cell migration inhibition experiment, a tube formation inhibition experiment, an in-vivo anti-angiogenesis activity experiment and the like show that the alantolactone can effectively resist angiogenesis. The application shows that the alantolactone has an inhibition effect on an MDA-MB-231 breast tumor and has a prospect of treating a breast cancer. A first research on a VEGFR2 and downstream signal passageway shows that the alantolactone can obviously inhibit phosphorylation of the VEGFR2 and also obviously inhibit phosphorylation of VEGFR2 downstream signal molecules including PLC(gamma)1, FAK, Src and AKT.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of inulin as an anti-angiogenesis drug. Background technique [0002] Inula is a perennial herb whose roots are used medicinally. It is cultivated in Europe and Tibet, China. Inula is mainly used in Europe for diuresis, expectorant and stomach invigoration, while in China it is mainly used for treating gastroenteritis, bronchitis and tuberculous diarrhea. Studies have shown that inulin contains a large amount of sesquiterpene lactones, and its main component, inulin, has strong antibacterial and anti-inflammatory effects. In recent years, studies on the pharmacological activity of inulin at home and abroad have found that inula lactones have the effect of anti-tumor cell proliferation, and their mechanism of inhibiting tumor growth has been studied. [0003] Alantolactone is a terpenoid compound, chemical name: 3a,5,6,7,8,8a,9,9a-octahydro-5,8a-dimethyl-3-meth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/365A61P35/00A61P35/02A61P17/06A61P19/02A61P9/10A61P9/00A61P15/14
CPCA61K31/365
Inventor 方超刘亚蓉赵梅陆琴
Owner SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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