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Application of transthyretin to inhibition of ocular neovascularization

A technology of thyroxine and angiogenesis, which is applied in the fields of biochemistry, molecular biology and medicine, can solve the problems of insensitivity and drug resistance of targeted drugs that have not been effectively overcome, and achieve the reduction and inhibition of ocular neovascularization The effect of new blood vessels

Inactive Publication Date: 2019-03-19
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 transthyretin to inhibition of ocular neovascularization
  • Application of transthyretin to inhibition of ocular neovascularization
  • Application of transthyretin to inhibition of ocular neovascularization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] like 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.

[0025] 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 microvascular ...

Embodiment 2

[0026] Embodiment 2 eye retinal endothelial cell migration experiment

[0027] 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 Cultivation of retinal 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, and wash the back of the chamber twice with PBS, 24 wells Add 600 μl of crystal violet to the plate, 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.

[0028] The result is as figure 2 As shown, the 48h cell migration test results show...

Embodiment 3

[0029] Example 3 Eye Retinal Endothelial Cell Tube Formation Experiment

[0030] Place the Matrigel stored at -20°C in a refrigerator at 4°C overnight, and it can be used when it completely melts into a red gelatinous liquid. Add Matrigel glue to the pre-cooled 48-well plate with a pre-cooled pipette tip, 150 μl per well. Gently shake the culture plate on ice to spread the Matrigel glue on the bottom of the plate. The 48-well plate was then placed in a 37° C. incubator for 30 min to solidify the Matrigel gel. When the cells grow to 80%-90% confluent, replace the serum-free medium and starve for 24h. Wash the cells 3 times with PBS, add 0.25% trypsin to digest the cells, the volume is limited to cover the bottom of the bottle, shake the bottle gently to make the trypsin fully contact with the cells, and incubate at 37°C for several minutes. Observe the cell morphology under an inverted microscope. When the cytoplasm is retracted and the intercellular space is enlarged, but t...

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Abstract

The invention discloses application of transthyretin (TTR) to the inhibition of ocular neovascularization and belongs to the field of biochemistry, molecular biology and medical science. The application finds out that mammal TTR is given and ocular new blood vessels in STZ (Streptozocin)-induced diabetes mellitus rat and mouse models are obviously reduced. An in-vivo experiment model of ocular neovascularization and brain neovascularization verifies that the TTR has the effect of inhibiting the neovascularization and a result shows that the TTR acts on ocular capillary endothelial cells and can be used for effectively inhibiting the formation of 90 percent of the blood vessels; and when the TTR acts in the rat and mouse models, high up to 90% of neovascularization can be effectively inhibited.

Description

technical field [0001] The invention relates to the application of transthyretin in inhibiting ocular 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...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P27/02A61P9/14A61P31/04A61P31/12A61P33/02A61P27/06A61P3/10A61P7/02A61P35/00A61P37/02A61P7/10
CPCA61K38/1709A61P3/10A61P7/02A61P7/10A61P9/14A61P27/02A61P27/06A61P31/04A61P31/12A61P33/02A61P35/00A61P37/02
Inventor 肖振
Owner SHANGHAI CUTSEQ BIOMEDICAL TECH CO LTD
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