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Functions and application of SHPS1 in treatment of post-vascular injury restenosis

A vascular injury and restenosis technology, applied in the field of gene function and application, can solve problems such as blood vessels affecting the treatment effect

Active Publication Date: 2014-12-03
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no cure for this kind of disease. The treatment methods of vascular surgery include balloon dilation, stent placement and arterial bypass, but restenosis after vascular reconstruction seriously affects the treatment effect

Method used

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  • Functions and application of SHPS1 in treatment of post-vascular injury restenosis
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  • Functions and application of SHPS1 in treatment of post-vascular injury restenosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Mouse Vascular Injury Model (VI) Obtained

[0037] 1. Grouping of experimental animals: WT and SHPS1-KO mice aged 8-10 weeks and weighing 24-27g were used and divided into two groups: WT vascular injury group and SHPS1-KO vascular injury group, with 20 mice in each group. mouse. The mice were sacrificed 28 days after the operation, and blood vessels in the injured segment were collected for analysis.

[0038] 2. Operation procedure of mouse vascular injury model:

[0039] 1) Accurately weigh the body weight (g) of the mouse in dynamic mode with an electronic balance, accurately prepare 3% pentobarbital sodium solution with double distilled water, shake gently to dissolve it fully, and use 80mg / kg body weight dose to calculate Accurately extract the corresponding volume of pentobarbital sodium solution with a 1mL syringe, and intraperitoneally inject the anesthetized mouse. After the mouse is fully anesthetized (about 3 minutes), 8% sodium sulfide is used to ...

Embodiment 2

[0044] Example 2 Determination of Intimal Neogenesis in Vascular Injury Model (VI) Mice

[0045] 1. Mice Harvesting

[0046] 1) Anesthetize the mouse, cut the heart and let the blood out.

[0047] 2) Cut the carotid artery from the proximal bifurcation of the carotid artery, take 0.5-0.6cm long, and keep the external carotid artery knot.

[0048] 3) Put the carotid artery in PBS, and gently drain the residual blood in the lumen with micro forceps.

[0049] 4) Put the blood vessel into a 1.5mL EP tube filled with 1mL 4% paraformaldehyde for fixation.

[0050] 2. Pathological detection

[0051] 2.1 Preparation of paraffin specimen slices

[0052] Paraffin specimen slices are prepared by professional pathologists in the laboratory. The main operating procedures include: after overnight fixation in 4% paraformaldehyde, carefully wrap the blood vessel with filter paper, put it into the embedding frame→rinse with running water→dehydration→transparency→wax immersion →embedding→s...

Embodiment 3

[0060] Example 3 Detection of proliferation level of blood vessel wall cells

[0061] The expressions of proliferating cell nuclear antigen (PCNA) and cell cycle protein (Cyclin D1) were detected by immunofluorescence staining. Required primary antibody information: PCNA (#2586; 1:100; mouse; Cell Signaling Technology), cyclin D1 (#2978; 1:25; rabbit; Cell Signaling Technology); required secondary antibody information: Alexa Fluor 568-conjugated goat anti-rabbit IgG (A11011; Invitrogen, Carlsbad, CA), Alexa Fluor 568-conjugated goat anti-mouse IgG (A11004; Invitrogen, Carlsbad, 150 d, CA).

[0062] The main steps are:

[0063] 1) Baked slices: put the paraffin slices in an oven at 55°C for more than 30 minutes.

[0064] 2) Dewaxing: Xylene 5min×3.

[0065] 3) Hydration: 100% ethanol 5min×2; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping for 5min×2.

[0066] 4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repair ...

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Abstract

The invention discloses functions and application of SHPS1 in treatment of post-vascular injury restenosis, belonging to the field of functions and applications of genes. SHPS1 gene knockout mice and wild type mice are taken as experimental subjects, and due to a vascular injury model, the mouse neointima formation, vascular wall cell proliferation level and smooth muscle cell phenotype transform are detected. The result proves that SHPS1 gene knockout can obviously promote neointima neogenesis and cell proliferation and promote transformation of smooth muscle cells from a shrinkage manner to a synthetic manner. Thus the functions of SHPS1 in post-vascular injury restenosis are realized, which mainly shows that SHPS1 has the functions of inhibiting neointima formation and cell proliferation and inhibiting smooth muscle cell phenotype transformation. Aiming at the functions of the SHPS1, the SHPS1 can be used for preparing medicines and arterial stents for preventing, relieving and / or treating the post-vascular injury restenosis.

Description

technical field [0001] The invention belongs to the field of gene function and application, and specifically relates to the function and application of SHPS1 in the treatment of restenosis after vascular injury, specifically the application of SHPS1 in the preparation of drugs for preventing, alleviating and / or treating restenosis after vascular injury. Background technique [0002] With the change of dietary structure and the aging process of the population, atherosclerotic occlusive lesions are increasing year by year and have become one of the main causes of death in our country. At present, there is no cure for this kind of disease. The treatment methods of vascular surgery include balloon dilation, stent placement and arterial bypass, but restenosis after vascular reconstruction seriously affects the treatment effect. Studies have shown that in the process of injury formation, the excessive proliferation of neointima and media tissue and the accompanying extracellular m...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61K48/00A61L31/16A61P9/10A61P9/00
Inventor 李红良王朗程文林张书敏王丕晓
Owner WUHAN UNIV
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