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Application of Toll interacting protein (Tollip) in stroke disease

A stroke and disease technology, applied in the application field of Toll-acting protein (Tollip) in stroke disease, to achieve the effect of protecting neurological function

Active Publication Date: 2014-08-20
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no international literature report on the application of Tollip in stroke diseases

Method used

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  • Application of Toll interacting protein (Tollip) in stroke disease
  • Application of Toll interacting protein (Tollip) in stroke disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] [Example 1] Acquisition of mouse cerebral infarction model (I / R)

[0030] 1. Grouping of experimental animals: male C57BL / 6 strain wild-type mice and Tollip gene knockout mice, cerebral infarction model (I / R) was established by middle cerebral artery ischemia-reperfusion. Randomly divided into 2 groups, 10 mice in each group: C57BL / 6 strain wild-type mouse I / R surgery group (WT I / R), Tollip gene knockout mouse I / R surgery group (KO I / R) .

[0031] 2. The I / R operation of cerebral infarction by suture method adopts the MCAO (middle cerebral artery occlusion, middle cerebral artery occlusion) model operation process:

[0032] (1) Grab the mouse, use 3% isoflurane to anesthetize the mouse, remove the mouse hair on the neck with 8% sodium sulfide, and quickly cut off the mouse hair on the top of the skull with surgical scissors, and disinfect the neck and skull with 3% iodine. Top skin twice, 75% alcohol deiodination once;

[0033] (2) Make a transverse incision on the sku...

Embodiment 2

[0037] [Example 2] Determination of cerebral infarction volume in cerebral infarction model (I / R) mice

[0038] 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.

[0039] (1) Neurological function and behavioral scores were performed 24 hours after operation and 72 hours before sampling;

[0040] Improved method based on Berderson neurological function score (9-point scale):

[0041] 0 points: no symptoms of nerve damage;

[0042] 1 point: The contralateral forelimb is curled up when the tail is raised, or the forelimb on the affected side cannot be fully reached;

[0043] 2 points: The opposite shoulder is adducted when the tail is raised;

[0044] 3 points: flat push: the resistance decreases when pushing to the opposite side;

[0045] 4 points: can move spontaneously in al...

Embodiment 3

[0059] [Example 3] Determination of apoptosis of neurons in the peripheral area of ​​brain tissue infarction

[0060] 1. Brain Tissue Cryosection Preparation

[0061] 1) Experimental mice were anesthetized by intraperitoneal injection of pentobarbital sodium at a dose of 50 mg / kg;

[0062] 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;

[0063] 3) Perfuse with 0.1mol / L PBS (pH7.4) at 100mmHg pressure until the liver becomes pale, and then perfuse with 4% paraformaldehyde for 15 minutes;

[0064] 4) Remove the mouse brain quickly after craniotomy, and fix it with 4% paraformaldehyde at room temperature for 6-8 hours;

[0065] 5) Resect the olfactory bulb and cerebellum of the brain tissue, then divide the brain into two parts successively along the midline, and then fix with the previous fixative for 15 minutes;

[0066] 6) Then immerse in phosphate buffer solution containing 30% su...

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Abstract

The invention discloses a function and application of a Toll interacting protein (Toll-interacting protein, Tollip) gene in a stroke disease, and belongs to the fields of functions and application of genes. The Tollip gene knockout mice are taken as experiment objects, and the model is perfused by middle cerebral artery occlusion. The result shows that the brain infarction volume of the Tollip gene knockout mice is obviously reduced in comparison with that of wild-type control mice, the neurological function is obviously improved, and the cell quantity of brain death is also obviously reduced, so as to find that the Tollip gene can promote deterioration of the nervous system, and aggravate and promote occurrence and development of the stroke. The Tollip can be used as a drug target for screening a drug of treating the stroke, and a Tollip inhibitor can be applied to preparation of the drug for treating the stroke.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and in particular relates to the application of a Toll action protein (Tollip) in stroke diseases. Background technique [0003] Ischemic stroke is currently the fourth leading cause of death and the second leading cause of disability in the world. Ischemic stroke can cause irreversible brain damage within minutes to hours after its onset, causing serious harm to the life and health of patients. Neuron cells are an important part of the central nervous system, but their high metabolic rate reduces the tolerance to ischemic and hypoxic environments, so they are more susceptible to damage than other neurovascular components. Currently, tissue plasminogen activator (tPA) fibrinolytic therapy is still the main treatment for ischemic stroke, but its time window of only 4.5 hours limits most patients to receive only symptomatic treatment. Studies have shown that neuronal protectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K45/00A61K48/00A61K39/395A61P25/00A61P9/10
Inventor 李红良郭森卢燕云郑安康李明昌
Owner WUHAN UNIV
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