Application of p53 activating agent in preparation of drugs for treating nerve injuries

A nerve injury and activator technology, applied in the field of medicine, can solve the problems of unclear axon regeneration of peripheral sensory neurons and central neurons

Inactive Publication Date: 2019-08-23
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether p53 can regulate the axon regeneration of pe

Method used

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  • Application of p53 activating agent in preparation of drugs for treating nerve injuries
  • Application of p53 activating agent in preparation of drugs for treating nerve injuries
  • Application of p53 activating agent in preparation of drugs for treating nerve injuries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: In vitro culture method of cortical neurons and hippocampal neurons

[0019] (1) Before cell plating, place sterile round glass slides in a 24-well plate in an ultra-clean bench and add 80.0 μL poly-D-lysine (100 μg / mL, Sigma-Aldrich) and laminin (10 μg / mL mL, Sigma-Aldrich) mixture coated round glass, after that, put the 24-well plate in 37 ℃, 5% CO 2 Approximately 2 hours in the cell culture incubator.

[0020] (2) The hippocampal neurons from E18 (embryonic day) were separated from other parts of the brain, and after removal, trypsin (Life Technology) was added and incubated in a metal bath at 37°C for 5 minutes.

[0021] (3) Immediately after trypsinization, wash three times with MEM medium containing 10% FBS to inactivate trypsin and terminate the digestion. The mixed hippocampal neuron cell suspension was used for cell counting, seeded in a 24-well plate at an amount of 30,000 / well, added medium (Neurobasal Medium, B27Supplement, GlutaMAX Supplemen...

Embodiment 2

[0023] Example 2: Detection of the regulatory effect of p53 activation on axon regeneration of sensory neurons

[0024] We added p53-specific activator Tenovin-6 (purchased from Selleck Chemicals) to DRG neurons cultured in vitro, and we found that activating p53 activity with Tenovin-6 significantly promoted axon regeneration of peripheral sensory neurons after 3 days of culture ( figure 1 A). In addition, we electroporated the mixture of pEX-3-p53 plasmid (synthesized in Shanghai Jima Pharmaceutical Technology Co., Ltd.) and pCAG-GFP plasmid (purchased in Matsuda & Cepko Company) into DRG neurons cultured in vitro. After 3 days of culture, we It was found that overexpression of p53 using the pEX-3-p53 plasmid effectively promoted the axon regeneration of sensory neurons ( figure 1 B).

[0025] At the same time, we electrotransfected the mixture of pEX-3-p53 plasmids and pCAG-GFP plasmids into L4-L5DRG neurons connected to the sciatic nerve. Two days after the transfection,...

Embodiment 3

[0026] Example 3: Detection of the regulatory effect of p53 activation on central neuron axon regeneration

[0027] We added Tenovin-6, a specific activator of p53, to E18 hippocampal neurons and E15 cortical neurons cultured in vitro. Through specific immunofluorescent staining of neurons, we found that Tenovin-6 activated p53 activity in hippocampal neurons. After that, axon regeneration in hippocampal neurons was significantly enhanced. Similar to the results in hippocampal neurons, the average length of axons in cortical neurons was significantly longer after activation of p53 activity ( figure 2 ). These results suggest that activation of p53 can promote axonal regeneration in central neurons. Thus, we determined that p53 is also an important regulator of axon regeneration in embryonic CNS neurons, and plays a positive regulatory role in axon regeneration of central neurons.

[0028] In addition, we will encode p53 type II adeno-associated virus vector (AAV2-p53, tite...

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Abstract

The invention discloses application of a p53 activating agent in preparation of drugs for treating nerve injuries, and belongs to the technical field of medicines. According to the application, it isdiscovered for the first time that p53 activation not only promotes axonal regeneration of peripheral sensory neurons, but also is an important regulatory factor for axonal regeneration of central neurons such as hippocampal neurons, cortical neurons and optic nerves. The discovery provides a new direction for treatment of patients with the nerve injuries such as spinal cord injuries.

Description

technical field [0001] The invention relates to the application of a p53 activator in the preparation of medicines for treating nerve damage, and belongs to the technical field of medicine. Background technique [0002] With the rapid development of modern economy, transportation and construction, the incidence of nerve injury, especially spinal cord injury, is also increasing year by year. According to statistics, the incidence rate of spinal cord injury reaches 40-60 per million people per year in China. Spinal cord injury often leads to permanent loss of sensory function, motor function and sphincter function below the level of injury, leading to paralysis and even death. According to the American Spinal Cord Injury Statistical Center, less than 1% of patients with spinal cord injury can fully recover their neurological function. In addition, the high medical expenses have brought a heavy burden to the patient's family and the whole society. However, there is currently ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/496A61P25/00
CPCA61K31/496A61K45/00A61P25/00
Inventor 赛吉拉夫马进进张浩楠齐士斌马艳霞张鸿程徐金辉秦绪祯
Owner SUZHOU UNIV
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