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Caspase activation and recruitment domain 3 (CARD3) and application of inhibitor of CARD3 to cerebral stroke diseases

An inhibitor, stroke technology, applied in disease diagnosis, nervous system disease, compound screening, etc., can solve the problems of short effective treatment time, unsatisfactory clinical treatment effect, etc., 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

The treatment of ischemic stroke mainly includes: thrombolysis, antiplatelet, anticoagulation, defibrosis, volume expansion, etc. However, due to the short effective treatment time window, the current clinical treatment effect is not satisfactory [2]

Method used

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  • Caspase activation and recruitment domain 3 (CARD3) and application of inhibitor of CARD3 to cerebral stroke diseases
  • Caspase activation and recruitment domain 3 (CARD3) and application of inhibitor of CARD3 to cerebral stroke diseases
  • Caspase activation and recruitment domain 3 (CARD3) and application of inhibitor of CARD3 to cerebral stroke diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] [Example 1] Acquisition of mouse cerebral infarction model

[0059] 1. Grouping of experimental animals: The cerebral infarction model in mice was obtained by middle cerebral artery ischemia-reperfusion (I / R). Animals were randomly divided into 4 groups, 10 mice in each group, which were control groups: non-transgenic littermate control I / R group (NTG I / R), wild type I / R group (WT I / R) And neuron-specific CARD3 transgenic I / R surgery group (CARD3-TG3), CARD3 gene knockout I / R surgery group (CARD3-KO I / R)

[0060] 2. Operation procedure of MCAO (middle cerebral artery occlusion, middle cerebral artery occlusion) model of cerebral infarction I / R surgery with thread embolization:

[0061] (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% active iodine Top skin 2 times, 75% alcohol d...

Embodiment 2

[0066] [Example 2] Determination of Cerebral Infarction Volume in Mouse Cerebral Infarction Model

[0067] 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.

[0068] (1) Neurological function and behavioral scores were performed 24h and 72h after surgery before sampling;

[0069] Improved method based on Berderson neurological function score (9-point scale):

[0070] 0 points: no symptoms of nerve damage;

[0071]1 point: The contralateral forelimb is curled up when the tail is raised, or the forelimb on the affected side cannot be fully reached;

[0072] 2 points: The opposite shoulder is adducted when the tail is raised;

[0073] 3 points: flat push: the resistance decreases when pushing to the opposite side;

[0074] 4 points: can move spontaneously in all directions, but ...

Embodiment 3

[0087] [Example 3] Determination of the apoptosis of neurons in the peripheral area of ​​brain tissue infarction

[0088] 1. Brain Tissue Frozen Section Preparation

[0089] (1) Grab the mouse and anesthetize the mouse by intraperitoneally injecting 3% pentobarbital sodium.

[0090] (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.

[0091] (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.

[0092] (4) The brain of the mouse was quickly taken out by craniotomy, and fixed with 4% paraformaldehyde at room temperature for 6-8 hours.

[0093] (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.

[0094] (6) Submerge in PBS (0.01M, pH 7.4) containing 30% sucrose, a...

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Abstract

The invention provides a Caspase activation and recruitment domain 3 (CARD3) and application of an inhibitor of the CARD3 to cerebral stroke diseases and belongs to the field of functions and application of genes. On one hand, an in-vitro cell experiment shows that adenovirus-mediated CARD3 interference can reduce injuries caused by anoxia / reoxygenation of primary cortical neuron cells and neuron cell apoptosis is inhibited; on the other hand, neuron-specific CARD3 knockout mice and neuron-specific CARD3 transgenic mice are used as experiment objects and a middle cerebral artery ischemia reperfusion model is used for researching; a result shows that the CARD3 has a neurological deterioration function and can aggravate and promote the development of cerebral stroke; theoretical basis and clinical foundation are provided for new targets and new strategies for research, prevention, alleviation and / or treatment of the cerebral stroke diseases.

Description

technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to the application of CARD3 as a drug target in screening drugs for treating ischemic stroke, and the application of CARD3 inhibitors in preparing drugs for treating ischemic stroke. 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 is mainly divided into two categories: ischemic stroke and hemorrhagic stroke, and ischemic stroke accounts for 60% to 80% of the total number of stroke patients. [3] . Ischemic stroke is the disorder of blood supply to local brain tissue caused by various reasons, leading to ischemic and hypoxic lesions and necrosis of brain tissue, which in turn leads to clinically corresponding neurological deficits. The pathophysiological changes of ischemic stroke mainly involve the following factors: excitotoxicity, o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573C12Q1/02A61K45/00A61K31/713A61K39/395A61K48/00A61P9/10A61P25/00
CPCG01N33/573A61K31/713A61K39/395A61K45/00A61K48/00G01N33/5058G01N2333/91215G01N2500/04G01N2800/2871
Inventor 李红良姬燕晓
Owner WUHAN UNIV
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