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Application of interferon regulatory factor 8 (IRF8) in cerebral apoplexy disease

A technology for stroke and disease, which is used in nervous system diseases, cardiovascular system diseases, medical preparations containing active ingredients, etc.

Active Publication Date: 2014-05-14
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A variety of neuroprotective drugs have achieved exciting results in animal experiments, but after entering the third phase of clinical trials for stroke, most of them failed to achieve the expected effect. One of the primary reasons for their failure is that most of the known The neuroprotective mechanism works within 4-6 hours after stroke, and it is difficult to implement treatment in such a short time window in clinical practice, so further elucidate the molecular mechanism that promotes or protects brain tissue damage for a long period of time after stroke occurs It is of great significance for the study of effective stroke treatment targets or strategies

Method used

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  • Application of interferon regulatory factor 8 (IRF8) in cerebral apoplexy disease
  • Application of interferon regulatory factor 8 (IRF8) in cerebral apoplexy disease
  • Application of interferon regulatory factor 8 (IRF8) in cerebral apoplexy disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] [Example 1] Construction of nerve-specific IRF8 transgenic mice

[0045] IRF8-flox transgenic mouse construction information:

[0046]Transgenic vector construction information: Use the upstream primer, namely 5'-CCAGATTACGCTGATTGTGACCGGAACGGCGGGCG-3' (SEQ ID NO. 1); the downstream primer, 5'-AGGGAAGATCTTGATTTAGACGGTGATCTGTTGAT-3' (SEQ ID NO. 2), to amplify the full length of mouse IRF8 Gene (NCBI, Gene ID: 15900, NM_008320.3), insert the cDNA into the pCAG-CAT-LacZ vector, which contains a CMV enhancer and a promoter of chicken β-actin gene (CAG, chicken β-actin gene) sub, and connected to the chloramphenicol acetyltransferase gene (CAT, chloramphenicol acetyltransferase), loxP sites are located on both sides of CAT. The expression of IRF8 in nerve cells is driven by the CAG promoter ( figure 1 A). IRF8-floxed mice: The constructed pCAG-IRF8-CAT-LacZ vector was constructed into fertilized embryos (C57BL / 6J background) by microinjection to obtain IRF8-floxed transgen...

Embodiment 2

[0050] [Example 2] Acquisition of mouse cerebral infarction model (I / R)

[0051] 1. Grouping of experimental animals: male C57BL / 6 strain wild-type mice, IRF8 knockout mice, brain-specific IRF8 transgenic mice and non-transgenic mice, cerebral infarction model was established by middle cerebral artery ischemia-reperfusion (I / R). Randomly divided into 8 groups, 10 mice in each group: C57BL / 6 strain wild type mouse sham operation group (WT SHAM) and I / R operation group (WT I / R), IRF8 gene knockout mouse sham operation group (KO SHAM) and I / R operation group (KO I / R), non-transgenic mouse sham operation group (NTG SHAM) and I / R operation group (NTG I / R), neuron-specific IRF8 transgenic mouse sham Surgery group (TG SHAM) and I / R surgery group (TG I / R).

[0052] 2. The I / R surgery of cerebral infarction by suture method adopts the mouse middle cerebral artery ischemia reperfusion (middle cerebral artery Ischemia Reperfusion) model operation process:

[0053] (1) Grab the mouse,...

Embodiment 3

[0059] [Example 3] Determination of Cerebral Infarction Volume in Cerebral Infarction Model (I / R) Mice

[0060] 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.

[0061] (1) Neurological function and morphology scores were performed 24 hours after operation and 72 hours before sampling;

[0062] Improved method based on Berderson neurological function score (9-point scale):

[0063] 0 points: no symptoms of nerve damage;

[0064] 1 point: The contralateral forelimb is curled up when the tail is raised, or the forelimb on the affected side cannot be fully reached;

[0065] 2 points: The opposite shoulder is adducted when the tail is raised;

[0066] 3 points: flat push: the resistance decreases when pushing to the opposite side;

[0067] 4 points: can move spontaneously in al...

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Abstract

The invention discloses function and application of an IRF8 gene in a cerebral apoplexy disease, belonging to the field of the function and the application of a gene. According to the invention, IRF8 gene knockout mice and neuron-specific IRF8 transgenic mice are taken as experimental subjects, a brain middle artery ischemia reperfusion model is adopted, and the results show that the cerebral infarction volume of the IRF8 gene knockout mice is obviously increased and the neurological function is obviously worsened as well in comparison with that of the wild C57 mice, the infarction volume of the IRF8 transgenic mice is obviously reduced, and the neurological function obviously gets better. The invention discloses the function of the IRF8 gene in the cerebral apoplexy disease, which mainly means that the IRF8 gene has an effect of protecting the function of a nervous system, especially an effect that the IRF8 gene can protect the cerebral apoplexy disease. According to the abovementioned function of the IRF8, the invention provides the application of the IRF8 in preparation of a drug for treating the cerebral apoplexy disease.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and particularly relates to the application of an interferon regulatory factor 8 (interferon regulatory factor 8, IRF8) in stroke diseases. [0003] Background technique [0004] Ischemic stroke is currently the leading cause of death and disability in the world. At present, tissue-type plasminogen activator (tPA) fibrinolysis is still the main treatment for ischemic cerebrovascular disease. Blood reperfusion will further aggravate ischemic nerve cell injury. Studies have shown that neuronal protection strategies can improve brain function and reduce neuronal cell loss long after cerebral ischemic injury. Apoptosis is one of the basic mechanisms of cell death during cerebral ischemia / reperfusion, but its regulatory mechanism is still not fully elucidated. Therefore, studying the molecular mechanism of neuronal apoptosis and survival during cerebral ischemia / reperfusion wil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61P25/00A61P9/10
Inventor 李红良郭森卢燕云蒋曦向梅张晓东
Owner WUHAN UNIV
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