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Interferon regulatory factor (IRF) 6 and application of IRF 6 inhibitor to brain stroke disease

An inhibitor, stroke technology, applied in cardiovascular system diseases, gene therapy, compound screening, etc., can solve problems such as unclear, and achieve the effect of worsening stroke disease

Inactive Publication Date: 2017-05-31
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether IRF6 regulates neurological diseases, especially ischemic stroke, in an immune-independent manner remains unclear

Method used

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  • Interferon regulatory factor (IRF) 6 and application of IRF 6 inhibitor to brain stroke disease
  • Interferon regulatory factor (IRF) 6 and application of IRF 6 inhibitor to brain stroke disease
  • Interferon regulatory factor (IRF) 6 and application of IRF 6 inhibitor to brain stroke disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] [Example 1] Acquisition of mouse cerebral infarction model

[0088] 1. Grouping of experimental animals: The cerebral infarction model in mice was obtained by middle cerebral artery ischemia-reperfusion (I / R). The animals were randomly divided into 4 groups, 10 mice in each group, respectively as the control group: non-transgenic littermate control I / R operation group (NTG I / R), IRF6 flox / flox I / R surgery group (IRF6 flox / flox I / R) and neuron-specific IRF6 transgenic I / R group (IRF6-TG4I / R), neuron-specific IRF6 knockout I / R group (IRF6-KO I / R)

[0089] 2. Operation procedure of MCAO (middle cerebral artery occlusion, middle cerebral artery occlusion) model of cerebral infarction I / R surgery with thread embolization:

[0090] (1) Grab the mouse, use 3% isoflurane to anesthetize the mouse, remove the mouse hair on the neck with 8% sodium sulfide, quickly cut off the mouse hair on the top of the skull with surgical scissors, and disinfect the neck and skull with 3% ...

Embodiment 2

[0095] [Example 2] Determination of Cerebral Infarction Volume in Mouse Cerebral Infarction Model

[0096] The evaluation indicators of the severity of cerebral ischemia / reperfusion injury mainly include cerebral infarct volume and neurological function score, and these indicators are positively correlated with the severity of ischemia / reperfusion injury.

[0097] (1) Neurological function and behavioral scores were performed 24h and 72h after surgery before sampling;

[0098] Improved method based on Berderson neurological function score (9-point scale):

[0099] 0 points: no symptoms of nerve damage;

[0100] 1 point: The contralateral forelimb is curled up when the tail is raised, or the forelimb on the affected side cannot be fully reached;

[0101] 2 points: The opposite shoulder is adducted when the tail is raised;

[0102] 3 points: flat push: the resistance decreases when pushing to the opposite side;

[0103] 4 points: can move spontaneously in all directions, but...

Embodiment 3

[0116] [Example 3] Determination of the apoptosis of neurons in the peripheral area of ​​brain tissue infarction

[0117] 1. Brain Tissue Frozen Section Preparation

[0118] (1) Grab the mouse and anesthetize the mouse by intraperitoneally injecting 3% pentobarbital sodium.

[0119] (2) Open the chest to expose the heart, puncture the left ventricle with an injection needle, and cut the right atrium at the same time.

[0120] (3) Perfuse with PBS (0.01M, pH 7.4) at 100 mmHg pressure until the liver turns white, and then perfuse with 4% paraformaldehyde for 15 min.

[0121] (4) The brain of the mouse was quickly taken out by craniotomy, and fixed with 4% paraformaldehyde at room temperature for 6-8 hours.

[0122] (5) The olfactory bulb and cerebellum of the brain tissue were removed, and then the brain was divided into two parts along the midline, and then fixed with the previous fixative for 15 minutes.

[0123] (6) Submerge in PBS (0.01M, pH 7.4) containing 30% sucrose, a...

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Abstract

The invention belongs to fields of gene functions and application. Neuron-specific IRF 6 knock-out mice and neuron-specific IRF 6 transgenic mice serve as experimental subjects, and a brain stroke model caused by middle cerebral artery ischemic reperfusion injury is researched, so results indicate that compared with wild mice, the IRF 6 gene knock-out mice have the characteristics that the infarct volume is remarkably reduced, the nerve function is remarkably improved, and the number of dead nerve cells is reduced; and IRF 6 gene overexpressed mice have the characteristics that the infarct volume is remarkably increased, the nerve function is remarkably deteriorated and the number of dead nerve cells is increased. In vitro cell experiments indicate that adenovirus-mediated IRF 6 interference can protect an injury of primary cortical neurons caused by anoxia / reoxygenation and inhibit neuron apoptosis. Therefore, the IRF 6 has the effect of deteriorating the nerve function, and can aggravate and promote development of brain stroke, and theoretical basis and clinical base are provided for researching a new target spot and a new strategy for preventing, relieving and / or treating the brain stroke disease.

Description

technical field [0001] The invention belongs to the field of gene function and application, and in particular relates to the application of IRF6 as a drug target in the screening of drugs for the treatment of ischemic stroke, and the use of IRF6 inhibitors in the preparation of drugs for preventing, alleviating and / or treating ischemic stroke application in medicine. Background technique [0002] Stroke is a sudden-onset cerebral blood circulation disorder that has become the leading cause of death and disability worldwide [1,2] . Stroke can be roughly divided into two categories: ischemic stroke, accounting for about 70% to 80%; hemorrhagic stroke, accounting for about 20% to 30% [3] . Ischemic stroke is permanent or temporary cerebral ischemic and hypoxic damage caused by thrombus or embolus blocking the great cerebral vessels or their branches. It is currently the fourth leading cause of death and the second leading cause of disability in the world. , irreversible bra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02A61K45/00A61K48/00A61K31/7105A61P9/10
CPCG01N33/5008A61K31/7105A61K45/00A61K48/005G01N2500/10
Inventor 李红良秦娟娟程文林
Owner WUHAN UNIV
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