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Functions and application of responsive to centrifugal force and shear stress gene 1 (RECS1) to treatment of restenosis after vascular injury

A technology for vascular injury and restenosis, which is applied to the function and application of centrifugal force and shear stress response gene 1 (RECS1) in the treatment of vascular injury and restenosis. Effect

Active Publication Date: 2014-12-10
武汉惠康基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is speculated that the expression of RECS1 gene may affect the function of vascular VSMC by regulating the TNF-α signaling pathway.

Method used

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  • Functions and application of responsive to centrifugal force and shear stress gene 1 (RECS1) to treatment of restenosis after vascular injury
  • Functions and application of responsive to centrifugal force and shear stress gene 1 (RECS1) to treatment of restenosis after vascular injury
  • Functions and application of responsive to centrifugal force and shear stress gene 1 (RECS1) to treatment of restenosis after vascular injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Mouse Vascular Injury Model (VI) Obtained

[0042] 1. Grouping of experimental animals: WT and RECS1-KO mice aged 8-10 weeks and weighing 24-27g were used and divided into two groups: WT vascular injury group and RECS1-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.

[0043] 2. Operation procedure of mouse vascular injury model:

[0044] 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

[0049] Example 2 Determination of Intimal Neogenesis in Vascular Injury Model (VI) Mice

[0050] 1. Mice Harvesting

[0051] 1) Anesthetize the mouse, cut the heart and let the blood out.

[0052] 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.

[0053] 3) Put the carotid artery in PBS, and gently drain the residual blood in the lumen with micro forceps.

[0054] 4) Put the blood vessel into a 1.5mL EP tube filled with 1mL 4% paraformaldehyde for fixation.

[0055] 2. Pathological detection

[0056] 2.1 Preparation of paraffin specimen slices

[0057] 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

[0065] Example 3 Detection of proliferation level of blood vessel wall cells

[0066] 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).

[0067] The main steps are:

[0068] 1) Baked slices: put the paraffin slices in an oven at 55°C for more than 30 minutes.

[0069] 2) Dewaxing: Xylene 5min×3.

[0070] 3) Hydration: 100% ethanol 5min×2; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping for 5min×2.

[0071]4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repair w...

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Abstract

The invention discloses functions and application of responsive to centrifugal force and shear stress gene 1 (RECS1) to treatment of restenosis after vascular injury. An RECS1 knockout mouse and a wild type mouse are taken as experimental subjects, and detection on mouse internal membrane regeneration, vessel wall cell proliferation level and smooth muscle cell phenotype transformation is carried out by a vascular injury model. Detection results prove that by virtue of RECS1 knockout, internal membrane regeneration and cell proliferation can be obviously inhibited, and smooth muscle cells are inhibited to be transformed into synthetic phenotype from contractile phenotype. Therefore, the functions of the RECS1 to the restenosis after vascular injury are mainly embodied in that internal membrane regeneration, cell proliferation and smooth muscle cell phenotype transformation are promoted by the RECS1. Aiming at the functions of the RECS1, the RECS1 can be used as a drug target for screening medicines for preventing, relieving and / or treating the disease of restenosis after vascular injury; an inhibitor of the RECS1 can be used for preparing medicines and arterial stents for preventing, relieving and / or treating the restenosis after vascular injury.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and specifically relates to the function and application of a centrifugal force and shear stress response gene 1 (responsive to centrifugal force and shear stress gene 1, RECS1) in the treatment of restenosis after vascular injury, specifically RECS1 Application in the preparation of drugs for preventing, alleviating and / or treating restenosis after vascular injury. Background technique [0003] 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. Neointimal formation is a key factor in atherosclerosis (AS), pulmonary hypertension, restenosis (RS) after percutaneous coronary intervention (PCI), post-transplant arterial disease, and pulmonary hypertension. Pathological process common to diseases. Vascular intimal neogenesis is...

Claims

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Application Information

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IPC IPC(8): G01N33/533A61K48/00A61K39/395A61P9/00A61L31/16
CPCA61K39/395A61K45/00A61K48/00A61L31/16A61L2300/432G01N33/533
Inventor 李红良赵辉张鹏张书敏王丕晓
Owner 武汉惠康基因科技有限公司
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