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Application of TTR (transthyretin) to angiogenesis inhibition

A technology of thyroxine and angiogenesis, applied in the fields of molecular biology, medicine, and biochemistry, can solve the problems of insensitivity and drug resistance of targeted drugs, and achieve the effect of inhibiting the generation of new blood vessels

Inactive Publication Date: 2019-03-08
SHANGHAI CUTSEQ BIOMEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, targeted drug anti-VEGF therapy is mainly used in the treatment of angiogenesis. However, there is no effective way to overcome the individual insensitivity and drug resistance of targeted drugs in clinical practice.

Method used

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  • Application of TTR (transthyretin) to angiogenesis inhibition
  • Application of TTR (transthyretin) to angiogenesis inhibition
  • Application of TTR (transthyretin) to angiogenesis inhibition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Such as figure 1 As shown, the TTR structure is centrosymmetric and axially symmetric; the monomer includes 147 amino acids, and the GeneID encoding TTR is 7276. TTR corresponds to the amino acid sequence of Genbank accession number CAG33189.1.

[0029] Under high glucose DMEM culture conditions, the cell viability of human microvascular endothelial cells (3000 / well) was detected by MTT assay with different concentrations of TTR. The results showed that after 48h, the cell proliferation of each group was compared, the A value of the 0μmol / L group was (0.40±0.03), and the A value of the 4μmol / L group was (0.17±0.02). The 4μmol / L group had a higher inhibitory effect on proliferation than the 0μmol / L group, and the difference was statistically significant (t=15.47, P=0.0001)( figure 1 ). Cell proliferation in the 4μmol / L group was reduced by 57.4% compared with the 0μmol / L group. It can be seen that TTR has an inhibitory effect on the proliferation of human microvascul...

Embodiment 2

[0030] Example 2 Migration experiment of umbilical vein endothelial cells

[0031] Add 600 μl of high-glucose DMEM medium containing 0 μM and 4 μM TTR to each well of a 24-well plate, and at the same time place a transwell chamber with a pore size of 8.0 μm, and add 200 μl of 1×10 4 Culture of human umbilical vein endothelial cells in FBS-free medium. After 48 hours, discard the medium in the upper and lower chambers, wipe the cells in the upper chamber with a cotton swab, add 600 μl of 0.4 g / L paraformaldehyde to the 24-well plate, fix at room temperature for 30 minutes, discard the paraformaldehyde, wash the back of the chamber twice with PBS, and use a 24-well plate Add 600 μl of crystal violet, stain at room temperature for 20 minutes, wash the back of the chamber twice with PBS after aspirating, observe and take pictures under a microscope, and randomly select 5 fields of view for cell counting.

[0032] The results of the 48h cell migration experiment showed that the nu...

Embodiment 3

[0033] Example 3 Cerebrovascular endothelial cell migration experiment

[0034] Add 600 μl of high-glucose DMEM medium containing 0 μM and 4 μM TTR to each well of a 24-well plate, and at the same time place a transwell chamber with a pore size of 8.0 μm, and add 200 μl of 1×10 4Culture of cerebrovascular endothelial cells in FBS-free medium. After 48 hours, discard the medium in the upper and lower chambers, wipe the cells in the upper chamber with a cotton swab, add 600 μl of 0.4 g / L paraformaldehyde to the 24-well plate, fix at room temperature for 30 minutes, discard the paraformaldehyde, wash the back of the chamber twice with PBS, and use a 24-well plate Add 600 μl of crystal violet, stain at room temperature for 20 minutes, wash the back of the chamber twice with PBS after aspirating, observe and take pictures under a microscope, and randomly select 5 fields of view for cell counting.

[0035] The result is as image 3 As shown, the results of the 48h cell migration e...

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Abstract

The invention discloses application of TTR (transthyretin) to angiogenesis inhibition, and belongs to the fields of biochemistry, molecular biology and medicine. After the TTR is given to mammals, inSTZ induced diabetes rat and mouse models, the new vessels of eyes are obviously reduced. Tumor new vessel, diabetes new vessel, eye new vessel and brain new vessel in vitro experiment models verify that the TTR has the effect of inhibiting the angiogenesis; the result shows that the TTR can act on an in vitro cell model in anoxic environment of brain and can continuously and efficiently inhibit the generation of new vessels. When the TTR acts on endothelial cells of umbilical veins, the formation of 90 percent of vessels can be obviously inhibited; when the TTR acts on endothelial cells of cerebral microvessel, the formation of 95 percent of vessels can be effectively inhibited; when the TTR acts on the endothelial cells of eye microvessel, the formation of 90 percent of vessels can be effectively inhibited; when the TTR acts on rat and mouse models, the as high as generation of 90 percent of new vessels can be effectively inhibited.

Description

technical field [0001] The invention relates to the application of transthyretin in inhibiting angiogenesis, and belongs to the fields of biochemistry, molecular biology and medicine. Background technique [0002] Angiogenesis refers to the formation of new blood vessels from the development of existing capillaries or postcapillary veins, mainly including: degradation of vascular basement membrane in the activation phase; activation, proliferation and migration of vascular endothelial cells; reconstruction of new blood vessels And the vascular network, is a complex process involving many kinds of molecules from many kinds of cells. Angiogenesis is a complex process of coordinated action of pro-angiogenic factors and inhibitory factors. Under normal circumstances, the two are in a state of balance. Once this balance is broken, the vascular system will be activated to cause excessive angiogenesis or inhibit the vascular system to degenerate blood vessels. The mechanism of ang...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P3/10A61P9/14A61P25/00A61P27/02A61P35/00A23L33/18
CPCA23L33/18A23V2002/00A61K38/1709A61P3/10A61P9/14A61P25/00A61P27/02A61P35/00A23V2200/30
Inventor 肖振
Owner SHANGHAI CUTSEQ BIOMEDICAL TECH CO LTD
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