Application of GABRD gene in screening drugs against heroin relapse

A heroin and gene technology, applied in the field of molecular biology, can solve problems such as no research reports yet

Pending Publication Date: 2020-10-23
宁波市微循环与莨菪类药研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been found that γ-aminobutyric acid A-type delta subunit receptor (GABRD) is involved in cocaine and alcohol addiction, but there is no relevant research report on the gene's role in heroin addiction and relapse at home and abroad.

Method used

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  • Application of GABRD gene in screening drugs against heroin relapse
  • Application of GABRD gene in screening drugs against heroin relapse
  • Application of GABRD gene in screening drugs against heroin relapse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Establishment of a stable rat heroin self-administration model

[0044] The classic rat heroin self-administration model can well simulate the heroin smoking behavior of clinical drug addicts. Rats can self-administer heroin after training, and the behavioral response rate and The dosage can reflect the behavioral development process of drug addicts from initial goal-directed drug use to compulsive habitual drug use.

[0045] We used the following methods to establish a self-administration model in rats: first, SD rats underwent jugular vein cannulation, a section of PE tube was inserted into the right jugular vein, and passed out of the body through the back, antibacterial and recovery for more than a week after the operation. Subsequently, the rats were trained in 32 self-administration operation cages. Before each training, the PE tube on the back of the rat was connected to the heroin injection system in the operation cage; there were two left and right na...

Embodiment 2

[0046] Construction of embodiment 2 overexpression vector

[0047]The present invention studies the effect of the GABRD gene on the relapse behavior of heroin addicted rats through two technical means of target gene overexpression and gene knockout. Overexpression of the target gene is one of the commonly used means to study gene function. The target gene is introduced into the cell by transfection (including electroporation, liposome transfection, etc.), microinjection or virus infection. In order to establish a stable cell line with overexpression of the target gene, it can be widely used in biological research, and RNAi technology can specifically knock out or turn off the expression of specific genes, so this technology has been widely used to explore gene functions and infectious diseases and malignant diseases. Therapeutic areas of cancer. The GABRD gene overexpression virus plasmid was constructed and transfected into 293T cells to establish a stable transfected cell l...

Embodiment 3

[0113] Example 3 Plasmid Transfection Cell Experiment

[0114] 1. Experiment overview

[0115] Cultivate the target cells in good growth state, that is, 293T cells. The day before plasmid transfection, divide the 293T cells into 24-well culture plates for culture. On the day of transfection, add the plasmids to the groups according to the experimental design for the transfection experiment of target cells. After 24 hours of transfection, the expression of GFP was observed under a fluorescent microscope, and the fluorescence rate was above 80%. After 48 hours of transfection, the cells were collected from the culture plate for detection.

[0116] 2. Experimental steps:

[0117] ①. Prepare target cells

[0118] (1) Take out the cell cryopreservation tube from the liquid nitrogen tank, quickly put it into a 37°C water bath, and shake it from time to time to thaw it as soon as possible;

[0119] (2) After complete thawing, centrifuge at 1300rpm for 3min, wipe and disinfect the ...

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Abstract

The present invention relates to an application of a GABRD gene in screening drugs against heroin relapse, and provides construction of a GABRD gene overexpression virus vector and a construction method of a GABRD gene interference adeno-associated virus vector. Compared with the prior art, the application of the GABRD gene in screening drugs against heroin relapse has the following advantages; aneffective therapeutic target for resisting against relapse and reducing psychological craving in heroin addiction is provided, and specific addition of expression of the GABRD gene in an important brain area of drug addiction (NAc) can significantly reduce drug-seeking behavior in heroin addiction. The results indicate that the the GABRD gene is expected to be a potential biomarker against heroinaddiction and relapse and a drug target for clinical treatment.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a GABRD gene and the application of the gene as a drug target in screening anti-heroin relapse drugs. Background technique [0002] Heroin addiction is a chronic relapsing brain disease with enormous social harm. Abuse affects many nervous systems in the brain, especially the neural circuits related to behaviors such as reward, motivation, learning and memory, and self-control, resulting in strong, uncontrollable drug craving and related compulsive drug-seeking behavior. It is involved in various physiological functions of organisms, and its dysfunction may be related to neuropsychiatric diseases such as schizophrenia, depression, epilepsy, dementia and other memory impairments. At present, more and more studies have shown that the GABAergic system plays an important role in the process of drug addiction and relapse. It can not only regulate the dopamine system but als...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/864C12N15/113C12N15/12G01N33/68
CPCC12Q1/6883C12N15/86C12N15/113C07K14/70571G01N33/6872G01N33/6893C12Q2600/136C12Q2600/158C12N2750/14143C12N2310/14G01N2333/70571G01N2800/307
Inventor 刘惠芬洪青晓徐文锦林姿陈为升赖苗军朱华强周文华
Owner 宁波市微循环与莨菪类药研究所
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