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Application of toll-acting protein (tollip) in stroke

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

Active Publication Date: 2017-02-22
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-acting protein (tollip) in stroke
  • Application of toll-acting protein (tollip) in stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] [Example 1] Mouse cerebral infarction model (I / R) was obtained

[0028] 1. Experimental animal grouping: male C57BL / 6 strain wild-type mice and Tollip gene knockout mice were used to establish cerebral infarction models (I / R) through middle cerebral artery ischemia-reperfusion. Randomly divided into 2 groups, each with 10 mice: C57BL / 6 strain wild-type mouse I / R group (WT I / R), Tollip gene knockout mouse I / R group (KO I / R) .

[0029] 2. The operation procedure of the MCAO (middle cerebral artery occlusion) model used in the I / R operation of thread embolization cerebral infarction:

[0030] (1) Grab the mouse, use 3% isoflurane to anesthetize the mouse, remove the rat hair from the neck with 8% sodium sulfide, cut off the rat hair quickly with surgical scissors, and sterilize the neck and skull with 3% vigor iodine Top skin 2 times, 75% alcohol deiodine 1 time;

[0031] (2) Make a lateral incision on the top of the mouse's skull to expose the skull, and gently peel off the con...

Embodiment 2

[0035] [Example 2] Measurement of cerebral infarct volume in cerebral infarction model (I / R) mice

[0036] The evaluation indicators of the severity of cerebral ischemia / reperfusion injury mainly include cerebral infarction volume and neurological function score, which are all positively correlated with the severity of ischemia / reperfusion injury.

[0037] (1) Perform neurological function and behavioral scores at 24h and 72h after surgery, respectively;

[0038] Based on Berderson neurological function score improvement method (9-point scale):

[0039] 0 points: no symptoms of nerve damage;

[0040] 1 point: The contralateral forelimb is curled up when the tail is lifted, or cannot reach the affected forelimb completely;

[0041] 2 points: Adduction of the opposite shoulder when lifting the tail;

[0042] 3 points: flat push: resistance drops when pushing to the opposite side;

[0043] 4 points: Can move spontaneously in all directions, but only turn to the opposite side when taking off t...

Embodiment 3

[0057] [Example 3] Determination of neuronal cell apoptosis in the peripheral area of ​​cerebral infarction

[0058] 1. Preparation of frozen sections of brain tissue

[0059] 1) Experimental mice are anesthetized by intraperitoneal injection of sodium pentobarbital at a dose of 50 mg / kg;

[0060] 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;

[0061] 3) After perfusion with 0.1mol / L PBS (pH7.4) 100mmHg until the liver becomes pale, perfusion with 4% paraformaldehyde for 15min;

[0062] 4) Take out the mouse brain quickly after craniotomy, and fix it for 6-8h after 4% paraformaldehyde at room temperature;

[0063] 5) Remove the olfactory bulb and cerebellum of the brain tissue, then divide the brain into two parts one after another along the median line, and fix it with the previous fixative for another 15 minutes;

[0064] 6) Submerge it in a phosphate buffer containing 30% sucrose and sink in a re...

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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

Technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to the application of a Toll action protein (Tollip) in stroke disease. Background technique [0002] Ischemic stroke is currently the fourth leading cause of death and the second leading cause of disability in the world. Irreversible brain damage occurs within a few minutes to a few hours after its onset, causing serious harm to the life and health of patients. Neuronal cells are an important part of the central nervous system, but their high metabolic rate reduces their tolerance to ischemic and hypoxic environments, so they are more susceptible to damage than other neurovascular components. At present, the treatment of tissue plasminogen activator (tPA) fibrinolysis is still the main treatment for ischemic stroke, but its only 4.5-hour time window limits most patients to receive symptomatic treatment. Studies have shown that neuron protection strategies can imp...

Claims

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Application Information

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Patent Type & Authority Patents(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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