Methods for constructing and identifying female AD model by utilizing receptor gene silencing technology

A technology of receptor gene and technical identification, applied in animal husbandry and other directions, can solve problems such as unsatisfactory treatment effect and inability to completely control disease development.

Inactive Publication Date: 2021-01-08
SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Therefore, the first-line drugs clinically used to treat AD are mainly cholinergic drugs, but their therapeutic effects are not satisfactory, only i

Method used

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  • Methods for constructing and identifying female AD model by utilizing receptor gene silencing technology
  • Methods for constructing and identifying female AD model by utilizing receptor gene silencing technology
  • Methods for constructing and identifying female AD model by utilizing receptor gene silencing technology

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Embodiment

[0044] Such as figure 1 Shown, a kind of method utilizing receptor gene silencing technology to construct female AD model, comprises the following steps:

[0045] (1) Randomly select 50 female rats and divide them into sham group, con-shRNA control group, and OXTR-shRNA group. The OXTR-shRNA group is divided into low-dose model group, middle-dose model group, and high-dose model group. Group of 10;

[0046] (2) After the animals were anesthetized, they were fixed on a rat brain stereotaxic instrument, and after routine scalp disinfection, the bregma was exposed and the position of the bregma was marked, and the coordinates of the basal forebrain were determined according to the "Rat Brain Stereotaxic Atlas" by Paxinos and Watson Point (AP: -1.0mm, L: ±2.7mm, V: 7.8mm) to adjust the drilling position, and use a three-edged needle to drill at the needle entry point;

[0047] (3) 0.2 μl of empty viral vector rAAV2-EGFP was injected into the basal forebrain of the con-shRNA cont...

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Abstract

The invention discloses a method for constructing a female AD model by utilizing an oxytocin receptor gene silencing technology and an identification method. Constructing and screening are carried outto obtain an AAV9-Oxtr-RNAi vector, AAV9-Oxtr-RNAi viruses are injected into a rat basal nucleus region by utilizing brain stereotaxis, according to a brain infection pathological state, detection and evaluation are carried out in combination with senile dementia symptoms caused by estrogen decline of clinical women, rats are randomly divided into a sham group, a con-shRNA control group and an OXTR-shRNA group, and the learning and memory abilities of the rats are detected by adopting a behavioral Morris water maze experiment and a bright and dark box experiment; an Elisa kit is used for detecting the change of the level of the rat brain tissue cortical estrogen (Estrogen, E2) and the change of the level of a acetylcholine neurotransmitter (acetylcholine, Ach); and in combination with animmunofluorescence technology, the co-localization and expression conditions of incoming nerve fibers of hippocampus in a brain basal nuclear cholinergic neuron fiber projection region, OTR and ChAT are observed, and a basis can be provided for better exploring and researching the pathogenesis and pathological changes of female Alzheimer disease.

Description

technical field [0001] The invention relates to the technical field of receptor gene silencing, in particular to a method for constructing and identifying female AD models using the receptor gene silencing technology. Background technique [0002] Alzheimer's disease (AD) is a chronic progressive neurodegenerative disease, which is a common type of dementia, and its incidence is increasing year by year. At present, the number of AD patients in my country has reached 6-10 million, which is the third leading cause of death in the elderly after cardiovascular diseases and tumors. Cholinergic damage is the core of the pathogenesis of AD, manifested as neuronal synaptic dysfunction, loss of pyramidal nerve cells, and massive degradation of acetylcholine, etc., which then cause and promote a series of pathological changes, and ultimately lead to complex and diverse pathological features and clinical features of AD. Performance. Therefore, the first-line drugs used in the clinica...

Claims

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Application Information

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IPC IPC(8): A01K67/027
CPCA01K67/0278A01K2217/206A01K2227/105A01K2267/0312
Inventor 侯雪芹荣翠平
Owner SHANDONG FIRST MEDICAL UNIV & SHANDONG ACADEMY OF MEDICAL SCI
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