Application of recombinant adeno-associated virus (AAV) expressing BDNF-HA2TAT in preparing medicaments treating depression diseases

A depression and virus technology is applied in the application field of recombinant adeno-associated virus in the preparation of drugs for treating depression, and can solve problems such as unreported

Inactive Publication Date: 2012-03-07
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But regarding the expression of BDNF-HA 2 The application of TAT's recombinant adeno-assoc...

Method used

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  • Application of recombinant adeno-associated virus (AAV) expressing BDNF-HA2TAT in preparing medicaments treating depression diseases
  • Application of recombinant adeno-associated virus (AAV) expressing BDNF-HA2TAT in preparing medicaments treating depression diseases
  • Application of recombinant adeno-associated virus (AAV) expressing BDNF-HA2TAT in preparing medicaments treating depression diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1 Construction of recombinant adeno-associated virus for packaging and expressing BDNF

[0028]Clone the BDNF cDNA fusion gene, connect the PCR product into the pGEM-T Easy vector, and obtain the pGEM-T Easy / BDNF transformed recipient strain E. Coli DH5α, the recombinant plasmid DNA was extracted by alkaline lysis, identified by enzyme digestion; the BDNF gene with cohesive ends was obtained by enzyme digestion, and the BDNF gene was inserted into the vector plasmid carrying the penetrating peptide TAT to construct the recombinant vector plasmid pSSCMV-BDNF-HA 2 TAT, recombinant plasmid pSSCMV-BDNF-HA 2 TAT was digested with EcoRI and BamHI to prepare BDNF-HA 2 The TAT fragment was connected to the PUC19 plasmid for sequencing; under the action of calcium phosphate, the recombinant vector plasmid pSSCMV-BDNF-HA 2 HEK293 cells were co-transfected with TAT, packaging plasmid pAAV / Ad and helper plasmid pFG140 to prepare and express BDNF-HA 2 Recombinant adeno-...

Embodiment 2

[0270] Example 2 Expression of BDNF-HA 2 Animal experiment of recombinant adeno-associated virus of TAT

[0271] Recombinant adeno-associated virus expressing brain-derived neurotrophic factor enters the brain through the nasal-brain pathway to produce antidepressant-like effects: C57 BL / 6 female and male adult mice were divided into nasal instillation of normal saline and BDNF-HA 2 TAT / AAV two treatment groups, the two treatments were divided into 1d administration group, 5 consecutive days administration group and 10 consecutive days administration group; the 1d administration group underwent behavioral study on 1, 2, and 3 days after administration respectively. Test (forced swimming test (forced swimming test, FST) and open field exploration test (open-field exploration test, OPF), 5 consecutive days of administration group and 10 consecutive days of administration group respectively at 1, 3, 7 days after administration Behavioral tests (FST and OPF) were performed ...

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Abstract

The invention relates to application of a recombinant adeno-associated virus (AAV) expressing BDNF-HA2TAT in preparing medicaments treating depression diseases. The invention has the advantages that: the recombinant AAV expressing BDNF-HA2TAT of the invention can have secretory expression of the macromolecular protein BDNF (brain derived neurotrophic factor), and the expressed BDNF can pass through a blood-brain barrier, and simultaneously introduction of a BDNF gene sequence into an AAV vector can generate continuous secretory expression of BDNF, thus solving the problem that BDNF needs repeated dosing; anti-depression efficacy screening experiments prove that the recombinant AAV able to have secretory expression of BDNF-HA2TAT has an anti-depression effect, and an approach and method for effective prevention and treatment of mental diseases like depressive disorders and stress disorders can be established.

Description

technical field [0001] The present invention relates to the application of a recombinant vector, specifically, a method for expressing BDNF-HA 2 Application of recombinant adeno-associated virus of TAT in preparation of medicine for treating depression. Background technique [0002] Depressive disorders have neuropathological changes such as hippocampal neuron atrophy, accelerated apoptosis, and neuronal regeneration disorders. How to effectively protect hippocampal neurons has always been a hot and difficult point in the study of depression. Brain derived neurotrophic factor (BDNF) is an important member of the neurotrophic factor family, and the down-regulation of BDNF may be involved in the structural and functional changes of the hippocampus in depression induced by chronic stress. Injecting exogenous BDNF into the hippocampal dentate gyrus of a depressed rat model of forced swimming and learned helplessness produced antidepressant-like activity, suggesting that centra...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K35/76C12N15/35C12N15/864A61P25/24A61K35/761
Inventor 马现仓高成阁党永辉杨广笑亢万虎伏炜王崴李雅妹马明芳
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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