Application of transcription factor Fo x O3 in preparing cerebral ischemia reperfusion nerve injury drug

A technology of cerebral ischemia reperfusion and transcription factors, applied in the field of medicine, can solve problems such as toxic side effects, bleeding complications, and weak curative effect

Inactive Publication Date: 2016-09-07
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the use of these drugs is time-limited and may lead to bleeding complications
The second type of therapy is neuroprotection, and most neuroprotective agents are relatively weak or have severe toxic side effects
Is FoxO3 transcription factor involved in the regulation of neuronal autophagy in cerebral ischemia-reperfusion injury? There is no literature report yet

Method used

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  • Application of transcription factor Fo x O3 in preparing cerebral ischemia reperfusion nerve injury drug
  • Application of transcription factor Fo x O3 in preparing cerebral ischemia reperfusion nerve injury drug
  • Application of transcription factor Fo x O3 in preparing cerebral ischemia reperfusion nerve injury drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Construction of adenoviral vector for overexpression of transcription factor FoxO3

[0060] 1. Cloning of FoxO3 target gene

[0061] The cDNA of human FoxO3 (gene sequence number NM_001455) was used as a template, and P1 was used as a primer (Table 1) to carry out PCR amplification of the target fragment, and the PCR amplification product was identified by 1% agarose gel electrophoresis, and a specific amplification product of about 2098bp was obtained. Increase the stripe size to match the size of the destination stripe ( figure 1 A); using the WT-FoxO3 gene as a template, respectively using P2, P3, P4, and P5 as primers (Table 1) to carry out PCR amplification of the target fragment to obtain specific amplification bands: 169bp fragment a, 666bp fragment b, fragment c of 225bp and fragment d of 1098bp, consistent with the size of the target band ( figure 1 B).

[0062] Table 1 Gene cloning primers

[0063]

[0064]

[0065] Specific amplified band...

Embodiment 2

[0096] Example 2: Effect of overexpression of FoxO3 on the expression of autophagy-related proteins in OGD / R injured HT22 cells

[0097] 1. Determination of the multiplicity of infection (MOI) value

[0098] The HT22 cells were infected with MOI values ​​of 50, 200, 500, and 1000. The results showed that when the MOI was 50, 200, 500, and 1,000, the infection efficiency was 50%, 75%, 85%, and 95%. The infection efficiency was the highest at 500 and 1000, but when the MOI was 1000, the cell damage was greater. On the basis of high infection efficiency, low MOI value, and low cytotoxicity, this experiment selected MOI values ​​between 200-500 for follow-up experiments ( Figure 9 ).

[0099] 2. Establishment of OGD / R model of HT22 cells

[0100] (1) Effect of OGD injury at different time on the activity of HT22 cells

[0101] MTT colorimetric assay was used to detect the effects of different periods of oxygen-glucose deprivation on the viability of HT22 cells. Taking the ce...

Embodiment 3

[0118] Example 3: Effect of overexpression of FoxO3 on the expression of autophagy-related proteins in rat MCAO / R injured brain tissue

[0119] 1. Establishment of MCAO / R model in rats

[0120] (1) Behavioral observation

[0121] Before cerebral ischemia, the vital signs of the rats were normal and they were awake. Loss of consciousness and mobility within 1 minute after ischemia, righting reflex disappears, limbs become stiff, pain disappears, bilateral pupils dilate, eyelid closure reflex disappears when stimulated by strong light at the same time, eye color is off-white (normally bright red) ). Immediately after reperfusion, the eyeball color returned to red and the pupil retracted. And after the rats woke up, Horner syndrome appeared on the right side, that is, the right eye fissure was small and the pupil was constricted; Turn right and fall down.

[0122] (2) TTC staining observation

[0123] The normal brain tissue was stained red, while the infarct after ischemia...

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Abstract

The invention relates to the technical field of medicine, in particular to application of a transcription factor Fo x O3 in preparing cerebral ischemia reperfusion nerve injury drug. The application of the transcription factor Fo x O3 in preparing the cerebral ischemia reperfusion nerve injury drug constructs two kinds of overexpression Fo x O3 plasmid, packs the overexpression Fo x O3 into the expression that adenovirus Ad-WT-Fo x O3 and Ad-TM-Fo x O3 regulates Fo x O3 and discusses the influence of Fo x O3 in OGD/R and MCAO/R damage on the expression of autophagy gap-associated protein LC3 and Beclin-1. According to experimental proof, Fo x O3 overexpression can promote the activation of autophagy gap-associated protein and has protective effect for the cerebral ischemia-reperfusion injury. The application of the transcription factor Fo x O3 in preparing the cerebral ischemia reperfusion nerve injury drug provides a new thought and a new path for the prevention of cerebral ischemic injury and a new target spot for the study of neuroprotective agents.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to the application of transcription factor FoxO3 in the preparation of a drug for protecting nerve injury from cerebral ischemia-reperfusion. Background technique [0002] Ischemic stroke is a common neurological disease characterized by high morbidity, disability and mortality (Hankey GJ, et al. Five-year survival after first-ever stroke and related prognostic factors in the Perth Community Stroke Study. Stroke, 2000, 31 (9), 2080-2086.), its prevention and treatment has become a key topic of concern to the whole society and medical circles. Studies have found that the key to the repair and functional recovery of brain tissue in patients with ischemic stroke is to rebuild blood flow (Yu J, et al. Activation of autophagy in rat brain cells following focal cerebral ischemia reperfusion through enhanced expression of Atg1 / pULK and LC3[J ]. Mol Med Rep, 2015, 12(3): 3339-3344.). How...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61P9/10A61P25/00
CPCA61K48/005A61K38/17
Inventor 陈霞吴晓燕周宏智刘爱芬
Owner NANTONG UNIVERSITY
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