Preparation method of composite animal model with Alzheimer disease

A technology of Alzheimer's disease and animal models, applied in the direction of active ingredients of aluminum/calcium/magnesium, which can solve the problem of not having the most suitable animal model for AD

Inactive Publication Date: 2011-05-18
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0039] The purpose of the present invention is to solve the problem in the prior art that there is no optimal AD animal model with all the characteristics of AD behavior and typical pathology, and to provide a preparation method for AD compound animal model

Method used

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  • Preparation method of composite animal model with Alzheimer disease
  • Preparation method of composite animal model with Alzheimer disease
  • Preparation method of composite animal model with Alzheimer disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Establishment of an Animal Compound Model of Aluminum Poisoning APP / PS1 Double Transgenic Mice

[0075] 16 healthy male C57BL / 6J mice and 16 healthy male APP / PS1 double transgenic mice (Institute of Zoology, Chinese Academy of Sciences) were selected, with a body weight of 20-22 g and similar mobility. They were randomly divided into 4 groups: wild mice control group, wild mice exposed to aluminum group, APP / PS1 double transgenic mice control group, APP / PS1 double transgenic mice AD ​​compound model group, with 8 mice in each group.

[0076] Freshly prepared 1mol / L AlCl 3 ·6H 2 O solution, after autoclaving, was exposed by intraperitoneal injection. The exposure period was 60 days, 0.05mL / d, injected every other day, and a total of 30 injections were injected within 60 days. Intramuscular injections of penicillin and streptomycin were given intermittently to combat infection. The control group was intraperitoneally injected with the same volume of sterilized normal s...

Embodiment 2

[0079] Morris water maze test

[0080] Morris water maze (Morris water maze, MWM) was performed after the poisoning. The Morris water maze consists of a white cylindrical pool and a transparent plexiglass platform that can be moved and adjusted in height. The circular pool has a diameter of 100cm, a height of 75cm, a platform height of 50cm, and a diameter of 10cm. The pool wall is composed of white metal plates. The water depth in the pool is 30cm, the platform is 1cm below the water surface, and the water temperature is 22±2°C. A camera with a display system is placed above the maze, and the computer automatically tracks the timing and records the swimming track. The external reference in the maze remained unchanged during the experiment.

[0081] Place navigation experiment: the mice received continuous training for 4 days, 4 times a day, with an interval of 20 minutes each time, and recorded the time required for the mice to enter the water from four different quadrants...

Embodiment 3

[0088]Preparation of tissue sections and routine HE staining

[0089] After the aluminum exposure period, the mice were anesthetized by intraperitoneal injection of pentobarbital sodium, perfused and fixed, and the brain was decapitated to remove the brain, and the removed cerebral cortex and hippocampal tissue were quickly fixed in 10% neutral buffered formalin, graded ethanol Dehydrated, cleared in xylene, embedded in paraffin, sectioned with RM-2245 paraffin microtome, thickness 5 μm, baked at 60°C for 12 hours, then taken out and stored at room temperature for routine and special histological staining.

[0090] 1) Dewax the tissue sections with xylene for 2×10 min; 2) Dexylene with absolute ethanol for 2×1~2 min; 3) 95%, 80%, and 70% ethanol for 2 min each, and wash with tap water for 1 min; 4) Hematoxylin staining for 3 min , washed with tap water for 1min; 5) 1% hydrochloric acid alcohol differentiation for 20s, washed with tap water for 1min; 6) 1% ammonia water turned ...

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Abstract

The invention relates to a preparation method of composite animal model with the Alzheimer disease. A healthy male APP (amyloid protein precursor)/PS1 (presenilin) double transgenic mouse is chosen, and a 1mol/LAlcl3.6H2O solution is used for intraperitoneal injection contamination for 30 times, and 0.05mL of the solution is injected once. After being detected by the neuroethology and the morphology, the composite animal model with the Alzheimer disease, which is obtained by the method, has obviously lowered learning and memory capability compared with animals in a control group and has the typical morphologic features, such as age pigments, neurofibril tangle, amyloid protein deposition, granulovacuolar degeneration and the like, can better simulate the neurobehavioral reaction of the Alzheimer disease and has all typical morphological changes of the Alzheimer disease. The composite animal model with the Alzheimer disease can be used for screening medicaments and researching the mechanism of the Alzheimer disease.

Description

technical field [0001] The invention belongs to the technical field of animal model construction, and relates to a preparation method of an Alzheimer's disease animal model, in particular to a preparation method of an aluminum-induced APP / PS1 double transgenic mouse Alzheimer's disease composite animal model. Background technique [0002] With the increase of human life expectancy, many countries have entered or are about to enter the aging society, and senile diseases have become a hot spot of social concern and related disciplines. Senile dementia (Alzheimer's disease, AD) is a chronic, progressive degenerative disease of the central nervous system. The rate increases year by year as the average life expectancy increases. Epidemiological surveys found that the incidence of AD in the elderly over 65 years old in my country is as high as 3.5%. [0003] Since the discovery of AD by German neuropathologist Alois Alzheimer in 1906, scientific workers have carried out painstak...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K33/06
Inventor 牛侨张勤丽贾丽教霞郭卫力
Owner SHANXI MEDICAL UNIV
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