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Application of PDLIM 1 in protection of cerebral ischemic neuron injury

A PDLIM1, 1. PDLIM1 technology, applied in the field of molecular biology and biomedicine, can solve problems such as failure

Pending Publication Date: 2020-11-03
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing drug candidates developed for a target in the mechanism of cerebral ischemic injury have failed clinical trials

Method used

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  • Application of PDLIM 1 in protection of cerebral ischemic neuron injury
  • Application of PDLIM 1 in protection of cerebral ischemic neuron injury
  • Application of PDLIM 1 in protection of cerebral ischemic neuron injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Establish a mouse model of cerebral ischemia-reperfusion injury (MCAO / R model):

[0024] The mice were weighed, fixed after intraperitoneal injection of 10% chloral hydrate anesthesia, the neck was cut midline, the subcutaneous tissue was separated with ophthalmic forceps, and the mouse trachea was exposed. The right common carotid artery (CCA) was first separated, and then the internal carotid artery ( ICA) and the external carotid artery (ECA) were separated, and attention should be paid to avoid damage to the vagus nerve during the separation process. From the bifurcation of the common carotid artery to the cephalic end, the proximal end of the right common carotid artery was ligated at the same time to make its main trunk free, then the right internal and external carotid arteries were separated, and the internal and external carotid arteries were fixed with non-injured arterial clips. Make an incision on the distal end of the external carotid artery near the ligati...

Embodiment 2

[0026] Establishment of HT22 cell line stably transfected with H-Sg PDLIM 1:

[0027] The mouse PDLIM 1 DNA sequence (Gene ID: mus NM-016861.4) was found on NCBI, and the gRNA fragment was designed using online design software (http: / / crispor.tefor.net). Send the designed fragments to Invitrogen for synthesis.

[0028] The experiment uses EPI CRISPR-Cas9 empty load. The experiment was divided into transfection empty vector group (CRISPR-Cas9 empty) and transfection plasmid group (CRISPR-Cas9-PDLIM 1).

[0029] The day before transfection, trypsinize HT22 cells, at a cell density of 2.5×10 5 Inoculate into 6-well culture plates, culture overnight, and the cell density should reach 90-95% confluency at the time of transfection.

[0030] DNA (4ug) and liposome2000 (10ul) were diluted to 50ul using serum-free opti-medium, mixed gently, and incubated at room temperature for 5min.

[0031] Mix the diluted DNA and diluted liposome2000 (total volume 100ul), mix gently, and incubat...

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Abstract

The invention discloses an application of PDLIM 1 in protection of cerebral ischemic neuron injury, and belongs to the field of molecular biology and biomedicine. The expression of PDLIM 1 in hippocampal neurons of mice is clarified for the first time by using brain sea horse tissues and hippocampal neuron HT22 cell strains of the mice. An MCAO / R injury mouse model and an OGD-induced neuron injurymodel are used for proving that PDLIM 1 expression in hippocampal neurons after ischemia injury is reduced for the first time. The PDLIM 1 gene in HT22 cells is knocked down, and proliferation of hippocampal neurons HT22 is inhibited.

Description

technical field [0001] The invention belongs to the fields of molecular biology and biomedicine, and specifically relates to the application of PDLIM 1 in protecting cerebral ischemic neuron damage. Background technique [0002] The statistics of the "China Stroke Prevention Report (2018)" pointed out that the incidence of stroke in my country has risen sharply, showing rapid growth among low-income groups, obvious gender and regional differences, and younger trends. Stroke prevention projects are urgent. Ischemic stroke accounts for about 80% of stroke patients. Ischemic stroke refers to the sudden decrease in blood perfusion of local blood supply arteries of brain tissue or complete interruption of blood flow supply, resulting in the destruction and disintegration of the cells. After ischemia, the blood supply, oxygen supply, and sugar supply to the brain tissue are reduced or completely stopped, causing extensive neuronal damage. The pathological mechanism of this dam...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/53
CPCG01N33/6893G01N33/53G01N2800/2871
Inventor 陈霞周雪莉
Owner NANTONG UNIVERSITY
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