Construction method and application of depression model

Abstract

The invention discloses a construction method and application of a depression model. A drug capable of activating neurons in the lateral habenular nucleus brain area to discharge is given to the lateral habenular nucleus area of the brain of the experimental animal, and the depression model is obtained. The invention develops a new method for establishing the depression animal model by utilizing drug induction, local drug administration is performed on the brain of an experimental animal, abnormal discharge activity of neurons of lateral habenula nucleus (LHB) is activated, depression-like behaviors are generated, and the depression-like behaviors are induced to be more stable. Therefore, a new and reliable depression animal model is provided for researching the brain mechanism of depression and researching and developing novel drugs and non-drug intervention methods.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine and relates to a method for constructing a depression model and its application. Background Art

[0002] Depression is the most common mood disorder. Currently, the cause of depression is unknown, and clinical antidepressant drugs have low efficacy, slow onset, large side effects, and a high recurrence rate after drug withdrawal. This makes it an urgent scientific problem to clarify the pathogenesis of depression, find effective targets, and study antidepressants with rapid, high-efficiency, and low side effects. To solve this problem, animal models that simulate depression are of great help. Animal models are experimental objects and materials with simulated manifestations of human diseases established in medical scientific research, used to study the occurrence, development laws, and prevention and treatment measures of human diseases.

[0003] In recent years, the research on the biological mechanism of depression has developed rapidly and has penetrated into the molecular and gene levels. Animal models of depression have also developed rapidly. Relatively classic animal models include the long-term chronic unpredictable stress model, hormone drug-induced model, inflammatory factor-induced model, social defeat model, restraint stress model, learned helplessness model, etc. For example, CN102405878A discloses a method for constructing an animal model of depression, creating a restraint-induced hyperlipidemic diet depression model for rats and mice. By simulating the etiology and symptoms of clinical depression in a depression model, rats are given a high-fat diet and restrained in a restraint cylinder for more than 4 hours every day to simulate the state of clinical depression patients who eat a lot of high-fat and high-energy foods and lack exercise. After modeling for more than 21 days, by measuring sucrose consumption and conducting behavioral tests such as open field test, spontaneous activity test, tail suspension test, and forced swimming test, as well as measuring the content of monoamine neurotransmitters in the brain, it is proved that this method successfully establishes a depression model. However, the pathogenesis of depression is still unclear, and the symptoms have a wide range and various types. Currently, existing animal models can only simulate a part of the depressive symptoms. Therefore, to clearly study the brain mechanism of depression and develop new therapeutic drugs and means, new animal models are still needed.

[0004] In summary, how to construct a depression animal model that meets the current needs of drug research and development, etc., is one of the urgent problems to be solved in the current field of depression research. Summary of the Invention

[0005] In view of the deficiencies of the prior art and actual needs, the present invention provides a method for constructing a depression model and its application, and develops a new method for establishing a depression animal model by drug induction.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a method for constructing a depression model, and the construction method includes:

[0008] Administering a drug capable of activating the neuronal discharge in the lateral habenula (LHb) region of the brain of an experimental animal to obtain a depression model.

[0009] In the present invention, by locally administering drugs to the brain of an experimental animal, depressive-like behaviors are produced, thereby providing a new and reliable animal model of depression for studying the brain mechanisms of depression, developing new drugs and non-drug intervention methods.

[0010] Preferably, the experimental animal includes any one of mice, rats, non-human primates, dogs, etc.

[0011] Preferably, the drug capable of activating the neuronal discharge in the lateral habenula region of the brain includes any one or a combination of at least two of potassium channel agonists, allosteric activators, GABA(A) receptor agonists and antagonists, or glutamate receptor agonists.

[0012] It can be understood that the drugs capable of activating the neuronal discharge in the lateral habenula region of the brain described in the present invention are well-known to those skilled in the art and obtained through conventional channels, such as commercially available luteolin, muscimol, diazepam, midazolam, etc.

[0013] Preferably, the method for localizing the lateral habenula region of the brain includes:

[0014] Performing three-dimensional imaging on the brain of an experimental animal by brain imaging technology, and based on the three-dimensional imaging, localizing the lateral habenula region of the brain by brain stereotaxic technology.

[0015] The method for constructing a depression model of the present invention has a wide range of applications and can be applied to various experimental animals such as rodents (the drug administration schematic diagram is as shown in Figure 1 shown) and non-human primates (the drug administration schematic diagram is as shown in Figure 2 shown).

[0016] Preferably, the administering method includes:

[0017] Implanting a drug delivery catheter in the lateral habenula region of the brain and administering the drug through the drug delivery catheter; or,

[0018] Using a microsyringe to inject and administer the drug in the lateral habenula region of the brain.

[0019] Preferably, the dosage of the drug capable of activating the neuronal discharge in the lateral habenula brain region is 0.001 mg / kg to 0.1 mg / kg (the volume is generally controlled at 5 to 10 microliters (μL)). For example, it can be 0.002 mg / kg, 0.003 mg / kg, 0.004 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.02 mg / kg, 0.05 mg / kg, 0.08 mg / kg, or 0.09 mg / kg, etc. The administration frequency is 1 to 2 times per day, and the administration period is 1 week to 2 weeks.

[0020] As a preferred technical solution, the method for constructing the depression model includes:

[0021] Three-dimensional imaging of the brain of the experimental animal is performed by brain imaging technology. Based on the three-dimensional imaging, the lateral habenula region of the brain is located and a drug administration catheter is implanted through brain stereotaxic technology. A drug capable of activating the neurons in the lateral habenula brain region is given to the animal through the drug administration catheter, or injection administration is performed in the lateral habenula region of the brain using a microsyringe. The dosage of the drug is 0.001 mg / kg to 0.1 mg / kg (the volume is generally controlled at 5 to 10 microliters (μL)). The administration frequency is 1 to 2 times per day, and the administration period is 1 week to 2 weeks, thereby obtaining a depression model.

[0022] In a second aspect, the present invention provides an application of a depression model obtained by using the method for constructing a depression model as described in the first aspect in determining a neural circuit suitable for relieving depression.

[0023] In a third aspect, the present invention provides an application of a neural circuit as a drug target in screening drugs for treating depression, wherein the neural circuit is determined by using a depression model obtained by the method for constructing a depression model as described in the first aspect.

[0024] Preferably, the screening includes determining whether a candidate drug can be used for preventing or treating depression based on the influence on the neural circuit before and after using the candidate drug.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The present invention develops a new method for establishing a depression animal model by drug induction. By locally administering drugs to the brain of the experimental animal, the abnormal discharge activity of the neurons in the lateral habenula (LHb) is activated, resulting in depressive-like behaviors, thereby providing a new and reliable depression animal model for studying the brain mechanism of depression, developing new drugs, and non-drug intervention methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1Schematic diagram of drug administration to the LHb brain region of rodents;

[0028] Figure 2 Schematic diagram of drug administration to the LHb brain region of non-human primates;

[0029] Figure 3 Figure showing the results of depression behavior detection. Detailed implementation manner

[0030] To further elaborate on the technical means and effects adopted by the present invention, the present invention will be further described below in conjunction with embodiments and drawings. It can be understood that the specific implementation manners described herein are only used to explain the present invention, rather than limiting the present invention.

[0031] For those not specifying specific techniques or conditions in the embodiments, follow the techniques or conditions described in the literature in this field or according to the product instructions. For reagents or instruments not specifying the manufacturer, they are all conventional products that can be obtained through regular channels.

[0032] Example 1

[0033] In this example, rats were used to construct a depression model. The specific experimental process included:

[0034] First, under standard anesthesia, the head skin of 8-week-old SD rats (purchased from Zhejiang Vital River Laboratory Animal Science Co., Ltd.) was incised, the skull was exposed and the surface of the skull was cleaned; then, three-dimensional imaging of the rat brain was performed. Referring to the rat brain atlas, a hole was drilled in the skull position directly above the LHb; the needle of a microinjector was slowly inserted into the LHb position of the brain using a stereotaxic apparatus, and the potassium channel agonist Luteolin (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany) was slowly injected into the LHb; the dosage of the drug was 0.001 mg / kg, the administration frequency was once a day, and the administration period was 1 week. The wound was sutured; an equal amount of physiological saline was injected for the experiment as a control group; after the rats recovered, behavioral tests were performed using the common classical forced swimming behavior paradigm, that is, the rats were placed in a circular acrylic bucket filled with water, and the swimming and floating immobile time of the rats was recorded as the behavioral index of depression.

[0035] The results were as Figure 3 shown. The rats induced by the potassium channel agonist Luteolin showed obvious depressive-like behaviors compared with the rats treated with physiological saline, manifested as a significant increase in the floating time, indicating that a depression model was successfully constructed.

[0036] In summary, this application has developed a new method for establishing an animal model of depression using drug induction. By locally administering drugs to the brains of experimental animals, abnormal discharge activities of neurons in the lateral habenula (LHb) are activated, resulting in depressive-like behaviors, thereby providing a new and reliable animal model of depression for studying the brain mechanisms of depression, developing new drugs, and non-drug intervention methods.

[0037] The applicant declares that the present invention uses the above embodiments to illustrate the detailed method of the present invention, but the present invention is not limited to the above detailed method, that is, it does not mean that the present invention must rely on the above detailed method to be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, and the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

Claims

1. A method for constructing a depression model, characterized in that, The construction method includes the following steps: Administering a drug that can activate the neuronal discharge in the lateral habenular nucleus region of the experimental animal's brain to obtain a depression model.

2. The method for constructing a depression model according to claim 1, characterized in that, The experimental animal includes any one of mice, rats, non-human primates, or dogs.

3. The method for constructing a depression model according to claim 1 or 2, characterized in that, The drug that can activate the neuronal discharge in the lateral habenular nucleus region of the brain includes any one or a combination of at least two of potassium channel agonists, allosteric activators, GABA(A) receptor agonists and antagonists, or glutamate receptor agonists.

4. The method for constructing a depression model according to any one of claims 1-3, characterized in that, The positioning method for the lateral habenular nucleus region of the brain includes: Performing three-dimensional imaging on the brain of the experimental animal through brain imaging technology, and based on the three-dimensional imaging, positioning the lateral habenular nucleus region of the brain through brain stereotactic technology.

5. The method for constructing a depression model according to any one of claims 1-4, characterized in that, The administration method includes: Implanting a drug delivery catheter in the lateral habenular nucleus region of the brain and administering the drug through the catheter; or Using a microsyringe to inject and administer the drug in the lateral habenular nucleus region of the brain.

6. The method for constructing a depression model according to any one of claims 1-5, characterized in that, The dosage of the drug that can activate the neuronal discharge in the lateral habenular nucleus region of the brain is 0.001 mg / kg to 0.1 mg / kg, the administration frequency is 1 to 2 times per day, and the administration period is 1 week to 2 weeks.

7. The method for constructing a depression model according to any one of claims 1-6, characterized in that, The construction method includes: Performing three-dimensional imaging on the brain of the experimental animal through brain imaging technology, and based on the three-dimensional imaging, positioning and implanting a drug delivery catheter in the lateral habenular nucleus region of the brain through brain stereotactic technology, and administering a drug that can activate the neurons in the lateral habenular nucleus region of the brain to the animal through the catheter, or using a microsyringe to inject and administer the drug in the lateral habenular nucleus region of the brain. The dosage of the drug is 0.001 mg / kg to 0.1 mg / kg, the administration frequency is 1 to 2 times per day, and the administration period is 1 week to 2 weeks to obtain a depression model.

8. Application of a depression model obtained by using the method for constructing a depression model according to any one of claims 1-7 in determining a neural circuit suitable for relieving depression.

9. Application of a neural circuit as a drug target in screening drugs for treating depression, wherein, The neural circuit is determined using the depression model obtained by the construction method of the depression model according to any one of claims 1-7.

10. The application according to claim 9, characterized in that, The screening includes determining whether a candidate drug can be used for preventing or treating depression based on the effects on the neural circuit before and after using the candidate drug.

Images