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Application of Ang-2 and genes thereof in pharmacy

A technology of drugs and inflammatory factors, applied in gene therapy, antipyretics, anti-tumor drugs, etc., can solve problems such as high disability and tissue infection, and achieve the effect of reducing rejection and reducing tumor mortality

Inactive Publication Date: 2011-07-13
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, lymphangioectatic lymphedema is often accompanied by frequent tissue infections in the limbs, the course of the disease develops rapidly, and the degree of disability is high

Method used

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  • Application of Ang-2 and genes thereof in pharmacy
  • Application of Ang-2 and genes thereof in pharmacy
  • Application of Ang-2 and genes thereof in pharmacy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Detection of related genes in chronic limb lymphedema tissue

[0033] Materials and methods

[0034] 1. Expression of Ang-2 mRNA and protein in human chronic lymphedema tissue

[0035] (1) Ang-2 mRNA extraction and detection method: take about 50 mg of human chronic lymphedema lower limb skin tissue, add 1 ml of Trizol, cut into pieces at 4°C, grind with a homogenizer, and collect the liquid into a 1.5 ml centrifuge tube. Add 400 μl of chloroform, mix thoroughly, and centrifuge; add an equal volume of isopropanol, and after centrifugation, add 1ml of 75% ethanol containing 0.01% DEPC to wash, centrifuge, discard the supernatant, blot the nozzle with filter paper, and air-dry at room temperature for 5 minutes. Add 20 μl of 0.01% DEPC water to dissolve RNA; detect OD with a nucleic acid analyzer 260 / OD 280 Ratio, to determine concentration and purity.

[0036] - Reverse transcription reaction (RT):

[0037] Prepare 20μl reaction system: 25mM MgCl 2 4μl, 10× revers...

Embodiment 2

[0055] Human skin lymphatic endothelial cells (LECs) and lymphatic defect animal model experiments

[0056] Materials and methods

[0057] 1. Isolation and culture of human skin lymphatic endothelial cells (LECs)

[0058] LECs were obtained from circumcised skin of young children. Specific method: cut the foreskin of the child after circumcision into 2×10mm 2 Long strips were digested with 2.4u / ml Dispase at 4°C overnight, and the epidermis was peeled off. Cut the leather into 1×2mm 2 After the size, put it in a 50ml centrifuge tube, add 0.1% collagenase (prepared with serum-free DMEM medium) and shake and digest at 37°C for 2-3 hours. After the digestion, the cell suspension was filtered through a 400-mesh metal filter, centrifuged at 1500 rpm for 5 minutes, the supernatant was discarded, and the cells were collected. LECs were obtained by immunomagnetic bead CD34 antibody negative sorting and CD31 antibody positive sorting, inoculated in a six-well plate bottomed with f...

Embodiment 3

[0111] Correlation experiment between lymphangioectatic lesions and increased expression of Ang-2 and inflammation in tissue 1. LECs cell proliferation experiment: The isolation and culture methods of LECs and BECs are the same as above. Using the same batch of foreskin isolation or cryopreserved LECs and BECs of the same generation, passed to the third generation can be used for cell proliferation and migration experiments. We first use inflammatory factors (IL-1α, IL-β, IL-6, TNFα, LPS (Cytolab)) to co-culture with the cells to observe their respective effects on cell proliferation, and then add Ang2 (R&D) or VEGF-C ( R&D), and then detect the changes of cells. Concrete method: tritium-labeled cell proliferation experiment-cell digestion, 96-well plate, 2000 cells / well, after inoculating for 4 hours (adhesion), change the EGM2 culture solution containing 1%BSA Add cytokines at the same time, dose Ang2 (200ng / ml), VEGF-C (100ng / ml), IL-1α (1ng / ml), IL-β (1ng / ml), IL-6 (1ng / ml...

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Abstract

The invention relates to the application of Ang-2, in particular to the application of Ang-2 in the pharmaceutical field. The technical proposal of the invention includes the application of Ang-2 and The invention relates to the application of Ang-2, in particular to the application of Ang-2 in the pharmaceutical field. The technical proposal of the invention includes the application of Ang-2 andthe genes thereof in preparing the medicine which can treat or prevent growth diseases of internal and external inflammatory lymphatic and the application of Ang-2 and the genes thereof in preparing tthe genes thereof in preparing the medicine which can treat or prevent growth diseases of internal and external inflammatory lymphatic and the application of Ang-2 and the genes thereof in preparing the medicine which can treat or prevent inflammatory diseases of organic lymphatic and surrounding tissues. The invention has the advantages of developing a new medical usage and a novel application ofhe medicine which can treat or prevent inflammatory diseases of organic lymphatic and surrounding tissues. The invention has the advantages of developing a new medical usage and a novel application of Ang-2 and the genes thereof.Ang-2 and the genes thereof.

Description

technical field [0001] The present invention relates to the use of Ang-2, in particular to the application in the field of pharmacy. Background technique [0002] Lymphatic neogenesis or regeneration is a common pathological change, seen in many diseases, such as chronic lymphedema, tissue trauma, organ transplantation, and tumor growth and metastasis. Since the regeneration mechanism of lymphatic vessels is still unknown, there are currently almost no treatments and control methods for diseases related to pathological lymphatic vessel growth. Therefore, elucidating the growth mechanism of lymphatic vessels is a clinical problem to be solved. [0003] Angiopoietins (Ang-1 and Ang-2) / Tie2 is a newly discovered growth factor family, which regulates the development and regeneration of blood vessels together with the VEGFs / VEGFRs family. Tie2 receptors are distributed on the surface of endothelial cells. Ang-1 and Ang-2 bind to the Tie2 receptor tyrosine kinase. Ang-1 is an ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/18A61K48/00A61P29/00A61P37/06A61P31/00A61P35/00A61P7/10
Inventor 刘宁飞胡学庆蒋朝华
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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