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Application of N-acetyl-D-glucosamine in preparation of drugs for preventing and treating Alzheimer's disease

A technology of glucosamine and acetyl group, applied in the application, drug combination, antipyretics and other directions, can solve the problem that the effect of neurodegenerative diseases has not been reported, and achieves improvement of cognitive memory impairment, inhibition of neuroinflammation, and improvement of cognition. effect of function

Active Publication Date: 2021-08-13
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the nervous system, it has only been reported to inhibit autoimmune encephalomyelitis, but its role in neurodegenerative diseases has not been reported yet

Method used

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  • Application of N-acetyl-D-glucosamine in preparation of drugs for preventing and treating Alzheimer's disease
  • Application of N-acetyl-D-glucosamine in preparation of drugs for preventing and treating Alzheimer's disease
  • Application of N-acetyl-D-glucosamine in preparation of drugs for preventing and treating Alzheimer's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The effect of Ogt deficiency on the activation of astrocytes in vivo and in vitro.

[0032] 1. Method

[0033] Ogt floxp / floxp Mice with Glast-Cre ERT2 Mice were crossed to produce inducible Ogt conditional knockout (cKO) mice. Male mice were injected with tamoxifen 100 mg / kg or control agent (intraperitoneal injection, once a day, for 5 consecutive days) at 7 weeks, and after 8 weeks A series of experiments were carried out with 3 rats in each group.

[0034]Isolation and culture of primary brain astrocytes from adult mice: Adult mice were dislocated cervically, quickly decapitated, dissected hippocampal and cortical tissues in pre-frozen PBS, and carefully peeled off the meninges under a stereomicroscope. After washing with cold PBS, the tissue was minced with a scalpel and then rotated with 0.125% trypsin at 37°C for 15 minutes. Slowly rotate with 1000 U / mg papain for 15 minutes at 37°C, and then mix well with 20 μl DNase I (50000 U / ml). The cell suspe...

Embodiment 2

[0043] Example 2 Can the activation of astrocytes cause inflammatory response after Ogt deficiency.

[0044] 1. Method

[0045] Enzyme-linked immunosorbent assay (ELISA): hippocampal tissue homogenate was lysed in ice buffer (500μl, 20mM Tris-HCl, 150mM NaCl, pH7.4, 1% Triton X-100, 1mM EDTA, 1x protease inhibitor tablet) Centrifuge at 14000g for 15min at 4°C, and collect the supernatant. 100 μl of cell and tissue supernatants were used for enzyme-linked immunosorbent assay of inflammatory factors IL-1β and TNF-α using an ELISA kit (Thermo Fisher Scientific).

[0046] Preparation of Ogt inhibitor (Ogt-I): Prepare Ogt inhibitor (5mM) stock solution with DMSO, add Ogt inhibitor to the medium with a final concentration of 20 μM, and co-culture with purified adult astrocytes for 72 hours, An equal volume of PBS was added as a control.

[0047] 2. Results

[0048] To determine whether activated astrocytes release inflammatory factors, IL-1β immunostaining was performed with cul...

Embodiment 3

[0051] Example 3 Effects of Activated Astrocytes on Hippocampal Neurons and Cognitive Function of Mice after Ogt Deficiency

[0052] 1. Method

[0053] Golgi staining of neurons and quantitative analysis of dendritic branches and spines: Golgi staining was performed according to the manufacturer's protocol (FD neurotechnologies, PK401). Briefly, fresh mouse brains were stored in Golgi-Cox solution in the dark for 14 days, and then incubated with 30% sucrose for 2 days at 4°C. Coronal sections were cut to 200 μm thickness with a cryostat. Sections were collected, dehydrated in absolute alcohol, washed in xylene, and covered with resin medium for mounting in sequence. Z-axis imaging of dendritic branches and dendritic spines of 6 to 8 pyramidal neurons in the CA1 region of each mouse was performed with an Olympus light microscope. Only cells with cell bodies in the section that were clearly stained, had no broken dendrites, and did not overlap adjacent cells were selected for...

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Abstract

The invention discloses application of N-acetyl-D-glucosamine (N-acetyl glucosamine, GlcNAc) in preparation of drugs for preventing and treating Alzheimer's disease, and particularly discloses an inhibiting effect of N-acetyl-D-glucosamine on astrocyte mediated neuroinflammation in O-GlcNAc transferase (Ogt) deficient mice and Alzheimer's disease model mice, and an improving effect on learning and memory disorder of astrocyte specific Ogt deficient mice, and researches the action mechanism of N-acetyl-D-glucosamine. The result shows that N-acetyl-D-glucosamine can significantly inhibit astrocyte mediated neuroinflammation in Ogt deficient mice or Alzheimer's disease model mice, and significantly improve the cognitive memory disorder of astrocyte specific Ogt deficient mice. The result shows that N-acetyl-D-glucosamine inhibits the inflammatory response and improves the cognitive function by improving the O-GlcNAc modification level of astrocytes, and can be used for preparing drugs or health food for preventing and treating Alzheimer's disease or other neuroinflammation related diseases.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and more specifically relates to the application of N-acetyl-D-glucosamine in the preparation of drugs for preventing and treating Alzheimer's disease. Background technique [0002] Astrocytes are the most abundant cell type in the CNS and regulate CNS functions such as maintaining CNS homeostasis and modulating synaptic information processing. Reactive astrocytes are present in certain pathological conditions, such as brain injury and neurodegenerative diseases. Reactive astrocytes secrete pro-inflammatory cytokines that affect neuronal survival, resulting in neurological disorders including Alzheimer's disease, multiple sclerosis, and spinal cord injury. Multiple mechanisms including epigenetic modifications and environmental factors regulate astrocyte function in central nervous system inflammation. [0003] O-GlcNAc transferase (Ogt) catalyzes the attachment of single-chain O-GlcNAc to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7008A61P25/28A61P29/00A23L33/125
CPCA61K31/7008A61P25/28A61P29/00A23L33/125A23V2002/00A23V2200/322A23V2250/61
Inventor 李学坤舒强董晓雪程雪军
Owner ZHEJIANG UNIV
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