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Application of differentiated embryonic cartilage development gene 1 in cerebral stroke disease

A stroke and gene technology, applied in the field of gene function and application, can solve the problems of unsatisfactory clinical treatment effect and short effective treatment time, and achieve the effect of inhibiting the occurrence of stroke and protecting neurological function

Inactive Publication Date: 2017-03-29
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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  • Application of differentiated embryonic cartilage development gene 1 in cerebral stroke disease
  • Application of differentiated embryonic cartilage development gene 1 in cerebral stroke disease
  • Application of differentiated embryonic cartilage development gene 1 in cerebral stroke disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] [Example 1] Acquisition of mouse cerebral infarction model

[0087] 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), DEC1 flox / flox I / R surgery group (DEC1 flox / flox I / R) and neuron-specific DEC1 transgenic I / R group (DEC1-TG3), neuron-specific DEC1 knockout I / R group (DEC1-KO I / R)

[0088] 2. Operation procedure of MCAO (middle cerebral artery occlusion, middle cerebral artery occlusion) model of cerebral infarction I / R surgery with thread embolization:

[0089] (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% act...

Embodiment 2

[0094] [Example 2] Determination of Cerebral Infarction Volume in Mouse Cerebral Infarction Model

[0095] 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.

[0096] (1) Neurological function and behavioral scores were performed 24h and 72h after surgery before sampling;

[0097] Improved method based on Berderson neurological function score (9-point scale):

[0098] 0 points: no symptoms of nerve damage;

[0099] 1 point: The contralateral forelimb is curled up when the tail is raised, or the forelimb on the affected side cannot be fully reached;

[0100] 2 points: The opposite shoulder is adducted when the tail is raised;

[0101] 3 points: flat push: the resistance decreases when pushing to the opposite side;

[0102] 4 points: can move spontaneously in all directions, but...

Embodiment 3

[0115] [Example 3] Determination of the apoptosis of neurons in the peripheral area of ​​brain tissue infarction

[0116] 1. Brain Tissue Frozen Section Preparation

[0117] (1) Grab the mouse and anesthetize the mouse by intraperitoneally injecting 3% pentobarbital sodium.

[0118] (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.

[0119] (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.

[0120] (4) The brain of the mouse was quickly taken out by craniotomy, and fixed with 4% paraformaldehyde at room temperature for 6-8 hours.

[0121] (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.

[0122] (6) Submerge in PBS (0.01M, pH 7.4) containing 30% sucrose, a...

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Abstract

The invention belongs to the field of gene functions and application. Neuronal specific DEC1 gene knockout mice and DEC1 gene overexpression mice are taken as experimental subjects, and by studying models that cerebral strokes are caused by middle cerebral artery ischemia reperfusion injuries of the mice, results show that compared with control mice, the infarct volume of the DEC1 gene knockout mice is obviously increased, the neurological functions are obviously deteriorated, the death nerve cell number is increased, the infarct volume of the DEC1 gene overexpression mice is obviously decreased, the neurological functions are obviously improved, and the death nerve cell number is decreased. In-vitro cell experiments show that by means of adenovirus-mediated DEC1 interference, hypoxia / reoxygenation injuries are prone to be generated on primary cultured cortical neuron cells, and apoptosis of the neuron cells can be promoted. Accordingly, the DEC1 has a neurological function protecting effect, can inhibit development of the cerebral strokes and can provide a theoretical basis and a clinical base for studying a new target and a new strategy which are used for preventing, relieving and / or treating the cerebral stroke disease.

Description

technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to the application of DEC1 as a drug target in screening drugs for treating ischemic stroke, and the application of DEC1 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, oxi...

Claims

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Application Information

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IPC IPC(8): A61K49/00A61K38/17A61P25/00A61P9/10
CPCA61K38/1709A61K49/0008
Inventor 李红良折志刚
Owner WUHAN UNIV
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