Method for constructing rat model for verifying property of drug for treating cerebral infarction ischemia

A technology of rat model and construction method, which is applied in the medical field, can solve problems such as poor therapeutic effect and inability to restore neurological function after cerebral ischemia, and achieve the effect of promoting recovery and improving therapeutic effect

Inactive Publication Date: 2018-04-13
FENGXIAN CENT HOSPITAL
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] In summary, the existing problems in the prior art are: the existing treatment for ischemic stroke cannot effectively promote the recovery of neurological function in the recovery period of cerebral ischemia, resulting in poor therapeutic effect; Due to the limitations of the protein harvesting technology of the target gene expression, there cannot be a certain abundance expression, so it is necessary to improve and hope for significant expression

Method used

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  • Method for constructing rat model for verifying property of drug for treating cerebral infarction ischemia
  • Method for constructing rat model for verifying property of drug for treating cerebral infarction ischemia
  • Method for constructing rat model for verifying property of drug for treating cerebral infarction ischemia

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Experimental program
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Effect test

Embodiment

[0071] 1. Isolation, culture and transfection of NSCs from neonatal rats were performed to analyze the expression of EPO gene.

[0072] 1. NSC cultivation

[0073] 1.1 Primary culture:

[0074] (1) Material collection: newborn SD suckling mice within 24 hours were killed by dislocation of the cervical spine, soaked in 75% alcohol for disinfection, the skull and meninges were removed, and the forebrain tissue was carefully separated under a dissecting microscope.

[0075] (2) Digestion: mechanical digestion is used. Cut the brain tissue sharply on a culture dish, transfer it to a 10ml cytocentrifuge tube, add PBS to 3ml, carefully pipette and let stand for 2min, suck out and collect the supernatant, add the same volume of PBS to the centrifuge tube containing the precipitated tissue liquid, pipetting again, and so on, so that the volume of the tissue gradually decreased.

[0076] (3) Centrifugation: Centrifuge at 75g for 10min (or 5min at 900 rpm), discard the supernatant. ...

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Abstract

The invention belongs to the medical field and discloses a method for constructing a rat model for verifying the property of a drug for treating cerebral infarction ischemia. NSC is separated from theforebrain tissue of a neonatal rat, the self-renewal and pluripotency differentiation characteristics are shown through in vitro culture and expansion by means of a flow cytometry and the single cellcloning technology, and through rAAV-mediated EPO gene and LacZ gene transfer, an NSC transfection system which stably expresses the EPO gene and the LacZ gene is established, and the in vitro and invivo transgene protein expression conditions of the NSC transfection system are analyzed; the NSC transfection system modified by the EPO gene and the LacZ gene is transplanted into cerebral infarction ischemia lesions of the rat, and the repairing effect of the EPO gene and the LacZ gene on the cerebral infarction ischemia lesions is analyzed. The EPO-modified NSC transplantation effectively promotes the recovery of the neurological function during the recovery from cerebral ischemia and improves the therapeutic effect.

Description

technical field [0001] The invention belongs to the medical field, and in particular relates to a method for constructing a rat model for verifying the properties of drugs for treating cerebral infarction and ischemia, and is particularly applicable to the analysis of the repairing effect of erythropoietin-modified neural stem cell transplantation on experimental rat cerebral infarction-ischemic lesions . Background technique [0002] Ischemic stroke is a general term for brain tissue necrosis caused by stenosis or occlusion of blood supply arteries (carotid artery and vertebral artery) and insufficient blood supply to the brain. There are four types of cerebral ischemia: transient ischemic attack (TIA); reversible neurological impairment (RIND); progressive stroke (SIE); and complete stroke (CS). TIA has no cerebral infarction, but RIND, SIE and CS have different degrees of cerebral infarction. Active medical treatment should be given to patients with cerebral ischemia in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/864C12N15/12C12N5/10A01K67/027
CPCA01K67/0275A01K2227/105A01K2267/035C07K14/505C12N5/0623C12N15/86C12N2510/00C12N2750/14143
Inventor 高山
Owner FENGXIAN CENT HOSPITAL
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