Function and application of transcription-induced spermatogenesis gene 40 (TISP40) in the treatment of restenosis after vascular injury

A technology for vascular injury and restenosis, which is applied in the field of gene function and application, can solve problems such as unsatisfactory curative effects, and achieve the effect of promoting restenosis

Active Publication Date: 2017-06-27
武汉惠康达科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the different degrees of mechanical damage during the operation can cause the formation of neointima, which leads to restenosis after stenting, and the curative effect is often not satisfactory.

Method used

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  • Function and application of transcription-induced spermatogenesis gene 40 (TISP40) in the treatment of restenosis after vascular injury
  • Function and application of transcription-induced spermatogenesis gene 40 (TISP40) in the treatment of restenosis after vascular injury
  • Function and application of transcription-induced spermatogenesis gene 40 (TISP40) in the treatment of restenosis after vascular injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Mouse Vascular Injury Model (VI) Obtained

[0036] 1. Grouping of experimental animals: WT and Tisp40-KO mice aged 8-10 weeks and weighing 24-27g were used and divided into two groups: WT vascular injury group and Tisp40-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.

[0037] 2. Operation procedure of mouse vascular injury model:

[0038] 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 t...

Embodiment 2

[0043] Example 2 Determination of Intimal Neogenesis in Vascular Injury Model (VI) Mice

[0044] 1. Mice Harvesting

[0045] 1) Anesthetize the mouse, cut the heart and let the blood out.

[0046] 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.

[0047] 3) Put the carotid artery in PBS, and gently drain the residual blood in the lumen with micro forceps.

[0048] 4) Put the blood vessel into a 1.5mL EP tube filled with 1mL 4% paraformaldehyde for fixation.

[0049] 2. Pathological detection

[0050] 2.1 Preparation of paraffin specimen slices

[0051] 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

[0059] Example 3 Detection of proliferation level of blood vessel wall cells

[0060] 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).

[0061] The main steps are:

[0062] 1) Baked slices: put the paraffin slices in an oven at 55°C for more than 30 minutes.

[0063] 2) Dewaxing: Xylene 5min×3.

[0064] 3) Hydration: 100% ethanol 5min×2; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping for 5min×2.

[0065] 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 the function and application of a transcription-induced sperm formation gene 40 (Tisp40) in the treatment of restenosis after blood vessel injury. In the present invention, Tisp40 gene knockout mice and wild-type mice are used as experimental objects, and the intimal neogenesis, proliferation level of blood vessel wall cells and phenotype conversion of smooth muscle cells are detected in the mice through the vascular injury model. The results showed that knockout of Tisp40 gene could significantly inhibit intimal neogenesis and cell proliferation, and inhibit the transition of smooth muscle cells from contractile to synthetic. This indicates that the function of Tisp40 in restenosis after vascular injury is mainly reflected in the promotion of intimal neogenesis, cell proliferation and smooth muscle cell phenotype conversion. For the above-mentioned functions of Tisp40, it can be used as a drug target for screening drugs for preventing, alleviating and / treating restenosis after vascular injury, and its inhibitors can be used to prepare drugs and arteries for preventing, alleviating and / treating restenosis after vascular injury. stand.

Description

technical field [0001] The invention belongs to the field of gene function and application, and specifically relates to the function and application of a transcription-induced sperm formation gene 40 (Tisp40) in the treatment of restenosis after vascular injury, specifically Tisp40 in the preparation of prevention, alleviation and / or treatment of vascular injury Drugs for restenosis. Background technique [0002] With the rapid development of human society and economy, the improvement of people's living standards and the acceleration of population aging process, the incidence of cardiovascular disease is increasing year by year, and it has become one of the major diseases that seriously endanger the global public health. The prevalence of cardiovascular disease in my country is on the rise, and about 3.5 million people die of cardiovascular disease every year, ranking first among all causes of death, causing great losses to society and the economy. Among cardiovascular dise...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K39/395A61K45/00A61K49/00A61P9/00
Inventor 李红良赵光年张鹏张书敏王丕晓
Owner 武汉惠康达科技有限公司
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