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Protective effect and application of tie2 on venous vessels in retina and other tissues

A technology of retinal veins and venous blood vessels, applied in the protective effect and application field of Tie2 on the venous blood vessels in the retina and other tissues, can solve the problem of ineffective regulation of blood vessel degree, and achieve the effect of controllable lesion degree

Active Publication Date: 2021-04-06
何玉龙
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the common disadvantage of these two models is that they cannot effectively control the degree of vascular abnormalities.

Method used

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  • Protective effect and application of tie2 on venous vessels in retina and other tissues
  • Protective effect and application of tie2 on venous vessels in retina and other tissues
  • Protective effect and application of tie2 on venous vessels in retina and other tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] The establishment of embodiment 1 conditional gene knockout mouse model

[0072] Use Tie2 + / - Mice mated with transgenic mice expressing UBC-CreERT2 or VE-Cadherin-Cre / ERT2 to obtain Tie2 + / - ; UBC-Cre / ERT2 or Tie2 flox / - ; VE-Cadherin-Cre / ERT2 mice, and then use this mouse with Tie2 flox / flox Mice mated to Tie2 flox / - ; UBC-Cre / ERT2 or Tie2 flox / - ; VE-Cadherin-Cre / ERT2 (also known as Tie2 in the present invention - / iKO ), mice of this genotype were used as experimental mice, and Tie2 obtained at the same time flox / + ; UBC-Cre / ERT2 or Tie2 flox / + ; VE-Cadherin-Cre / ERT2 was used as a control mouse (Control).

Embodiment 2

[0073] Example 2Tie2 gene knockout and detection of its knockout efficiency

[0074] The experimental mice obtained in Example 1 and the control group were subjected to the following experiments respectively, that is, the Tie2 gene knockout was induced in the following stages:

[0075] (1) From the first day of birth (P1) to the fourth day (P4), each mouse was injected (intragastrically) with 60ug of tamoxifen every day for 4 consecutive days;

[0076] (2) From the 5th day (P5) to the 8th day (P8), each mouse was injected (intragastrically) with 100ug of tamoxifen every day for 5 consecutive days;

[0077]The mouse in Example 1 was induced to knock out the gene after birth, and the lung tissue was analyzed on the 7th day. Extract the lung tissue of the induced transgenic mouse, add the tissue lysate containing protease inhibitors to fully homogenize and lyse, centrifuge, add the supernatant solution, add the sample buffer and boil for 15 minutes, then perform polyacrylamide ...

Embodiment 3

[0079] Example 3 Tie2 Gene Knockout and Detection of Retinal Vein Vascular Lesions in Retinal Tissue

[0080] The mouse in Example 1 was induced to knock out the gene after birth, and the retinal blood vessels were analyzed on the 7th day. Mouse eyeballs were removed and observed by immunofluorescence, Tie2 - / iKO Compared with the littermate control mice, the preretinal vein vessel growth was significantly increased ( figure 2 A), the quantitative results of blood vessel density at the venous end and arterial end are as follows figure 2 As shown, the vascular density of the blood vessels around the venous end increased ( figure 2 B), while the vessel density around the vessels at the retinal artery end did not change ( figure 2 C),. And we also observe that Tie2 - / iKO The mice had abnormal morphology of retinal veins and degeneration of vein characteristics, accompanied by increased angiogenesis around the veins (capillary neogenesis was indicated by the molecular mar...

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Abstract

The present invention relates to the protective effect and application of Tie2 on veins and blood vessels in the retina and other tissues, and specifically relates to the use of Tie2 regulating compounds in the preparation of medicines for the treatment of veins and related vascular diseases in tissues and organs such as the retina, skin, liver and lungs. use. The invention also relates to a conditional tyrosine kinase receptor Tie2 gene knockout non-human animal model closely related to the degree of venous vascular lesions in retina and other tissues and its preparation method, and its use in screening Tie2 regulatory compounds. By combining different doses of tamoxifen and different stages of animal growth, non-human animal models with different Tie2 gene knockout efficiencies can be obtained to simulate different degrees of venous vascular lesions in retinal and other tissues, and solve the problem of retinal and other tissues. The problem of the controllability of the degree of venous vascular disease.

Description

technical field [0001] Use of the Tie2 regulating compound of the present invention in the preparation of medicines for treating venous and related vascular lesions in tissues and organs such as retina, skin, and liver. The present invention also relates to a non-human transgenic animal model and its construction method and application, in particular to a controllable retinal and other tissue venous vascular disease Tie2 gene knockout induced by different doses of tamoxifen at different growth stages In addition to the transgenic mouse animal model and its construction method. Background technique [0002] Vein-related diseases include venous thrombosis, varicose veins, and venous inflammation. At present, little is known about the pathological process of clinical venous vascular disease, and the treatment options are limited. Therefore, the preparation of related disease models has important scientific value and application significance for elucidating the mechanism of ve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K49/00A61K31/138A61P9/10A61P9/14A01K67/02
CPCA01K67/02A01K67/0276A01K2217/075A01K2217/15A01K2227/105A01K2267/03A61K45/00A61K49/0008A61P1/16A61P9/10A61P9/14A61P11/00A61P17/00A61P27/02A61P29/00
Inventor 何玉龙储蔓李桃桃
Owner 何玉龙
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