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Cell culturing extract for degrading amyloid beta, preparation method and application thereof

A technology of amyloid protein and cell culture, which is applied in the field of preparation of drugs for the treatment of Alzheimer's disease, can solve the problems of no neurotoxicity, achieve the effect of improving cognitive ability and reducing amyloid deposition

Inactive Publication Date: 2009-10-07
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite the presence of amyloid throughout the brain, some brain regions lack significant amyloid deposition and the resulting neurotoxicity

Method used

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  • Cell culturing extract for degrading amyloid beta, preparation method and application thereof
  • Cell culturing extract for degrading amyloid beta, preparation method and application thereof
  • Cell culturing extract for degrading amyloid beta, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1) Experimental animals

[0069] APP / PS1 transgenic mice whose genetic background is C57BL / 6J, wild-type littermates and C57 mice were purchased from the Chinese Academy of Medical Sciences.

[0070] 2) Isolation and culture of primary neural cells in non-Alzheimer's disease lesion area of ​​mice and preparation of cell culture extracts

[0071] One day in advance, the bottom was laid with 500 μg / ml poly-D-lysine (Sigma-Aldrich). Prepare planting solution (80% DMEM + 10% fetal bovine serum + 10% horse serum + glutamine 500μM), digestive solution (0.25% trypsin) and prepare 5 discs of dissection solution (glucose 0.3%, sucrose 0.75%, Hepes 0.23%, NaCl 0.9%, KCl 0.04%, NaCl 2 HPO 4 ·7H 2 O 0.018%, KH 2 PO 4 0.003%, phenol red 0.12‰), a total of 6 culture dishes. The 18-day gestational C57 mice were sacrificed by neck joint dislocation, and the brainstem, pons, basal ganglia, subcortical white matter, cerebellum, spinal cord, and oblongata were removed from the fet...

Embodiment 2

[0114] 1) Experimental animals

[0115] SD rats were purchased from the Experimental Animal Center of Peking University Health Science Center. APP / PS1 transgenic mice with a genetic background of C57BL / 6J and wild-type littermates were purchased from the Chinese Academy of Medical Sciences.

[0116] 2) Isolation and culture of primary nerve cells in non-Alzheimer's disease lesion area of ​​rats and preparation of cell culture extracts

[0117] One day in advance, the bottom was laid with 500 μg / ml poly-D-lysine (Sigma-Aldrich). Prepare planting solution (80% DMEM + 10% fetal bovine serum + 10% horse serum + glutamine 500μM), digestive solution (0.25% trypsin) and prepare 5 discs of dissection solution (glucose 0.3%, sucrose 0.75%, Hepes 0.23%, NaCl 0.9%, KCl 0.04%, NaCl 2 HPO 4 ·7H 2 O 0.018%, KH 2 PO 4 0.003%, phenol red 0.12‰), a total of 6 culture dishes. The 18-day gestational SD rats were sacrificed by neck joint dislocation, and the brainstem, pons, basal gangli...

Embodiment 3

[0134] Injection of cerebrospinal fluid extracts in laboratory animals and diseased parts of laboratory animals

[0135] Rabbits were purchased from the Animal Center of Capital Medical University. APP / PS1 transgenic mice with a genetic background of C57BL / 6J and wild-type littermates were purchased from the Chinese Academy of Medical Sciences. The injection of the cell culture extract was carried out (0.8 mm posterior to bregma, 2.5 mm subdural, and 0.8 mm lateral) with a brain stereotaxic instrument (NARISHIGE, Scientific Instrument Lab, Tokyo, Japan) referring to the standard atlas (Academic Publish). The injection time of each animal was 5 minutes, and the needle was retained for 10 minutes. Seven days after the injection, the behavioral experiments were performed, and then the brain tissue was extracted for histological, morphological and biochemical detection.

[0136] Except for the isolation and culture of primary nerve cells and the preparation of cell culture extra...

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Abstract

The invention relates to a cell culturing extract for degrading amyloid beta, a preparation method, and application of the cerebrospinal fluid extract for preparing a medicament for treating alzheimer's disease. Concretely, the cell culturing extract can not only reduce amyloid deposition in in brain and neuro toxic reaction like loss of nerve synapse and glial cell hyperplasia, but also substantially enhance cognition capability of alzheimer's disease model animals.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular to the field of drugs for treating Alzheimer's disease. Specifically, the present invention relates to a cell culture extract for degrading β-amyloid protein, a preparation method thereof, and an application of the cell culture extract as a drug for treating Alzheimer's disease. Background technique [0002] Alzheimer's disease is a neurodegenerative disease with complex pathogenesis and still incurable. Among the many hypotheses about the pathogenesis, the amyloid hypothesis is currently the most accepted. According to this hypothesis, the most potentially valuable therapeutic approach is to prevent and treat the Alzheimer's disease process through anti-amyloid therapy. In fact, some anti-amyloid drugs have been proposed and even used in clinical treatment, such as inhibitors of secretase, a key protein in the process of generating amyloid. [0003] Alzheimer's disease is chara...

Claims

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Application Information

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IPC IPC(8): A61K35/30A61K35/24A61P25/28
Inventor 王钊杜婧孙兵张朗
Owner TSINGHUA UNIV
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