A method for establishing an animal model of alcoholic depression and its application

Abstract

The invention discloses a method for establishing an alcoholic depression animal model, comprising the following steps: selecting experimental animals that meet experimental requirements, and gavaging the experimental animals with a preset amount of acute alcohol for multiple times for pretreatment; after the acute alcohol pretreatment, LPS is used to induce the experimental animals until depression-like behavior occurs, and the constructed alcoholic depression model is verified; the preset gavage amount is calculated based on the experimental animal weight, with 5-10 g / kg for each experimental animal, preferably using the gavage of LPS for induction, which is more in line with the actual situation of alcoholic depression, and is calculated based on the experimental animal weight, with 10-15 mg / kg for each experimental animal; using the method, depressive behavior can be induced within 2-4 days, and compared with the chronic alcohol induction cycle, the method greatly shortens the experimental modeling time, and the mouse depressive behavior is significant, and the constructed alcoholic depression model has a good effect.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and more specifically, relates to a method for establishing an alcoholic depression animal model and an application thereof. Background Art

[0002] Depression is a common mental disorder, with typical symptoms including low mood, slow thinking, loss of interest, eating and sleeping disorders, cognitive impairment and other physical symptoms. In recent years, the incidence of depression has continued to rise, with at least 800,000 people with depression committing suicide each year. The incidence of depression in a person's lifetime is 12%-17%, and alcohol-induced depression can easily form a vicious cycle, which in turn aggravates the symptoms of depression.

[0003] Despite the grim situation, we still do not fully understand the etiology and pathogenesis of alcoholic depression. The main reason is that most of the existing depression models are different from the actual situation of alcoholic depression. In order to fully understand the pathogenesis of alcoholic depression or develop anti-alcoholic depression drugs, it is a key step to construct different alcoholic depression animal models in a targeted manner. However, the existing alcoholic depression models are relatively single. In the study of the relationship between alcohol and depression, it was found that chronic alcohol long-term treatment can induce depressive-like behavior, while acute alcohol treatment alone is difficult to induce depressive behavior, but the induction period of chronic alcohol is long, and there is currently no animal model that can fully cover all symptoms of depression.

[0004] Therefore, exploring a rapidly induced alcohol-induced depression model will help to fully understand the pathogenesis of alcohol-induced depression and promote research on the potential mechanisms of the relationship between alcohol and depression or anti-alcohol depression drugs. Summary of the invention

[0005] In view of the above defects or improvement needs of the prior art, the present invention provides a method for establishing an alcoholic depression animal model and its application, the purpose of which is to first use alcohol gavage combined with LPS to induce an alcoholic depression model, thereby solving the technical problems of the existing alcoholic depression model with a long induction period and a single model type.

[0006] To achieve the above object, according to one aspect of the present invention, a method for establishing an alcoholic depression animal model is provided, which comprises the following steps:

[0007] Select experimental animals that meet the experimental requirements;

[0008] The experimental animals are pretreated by multiple intragastric administration of a preset amount of acute alcohol; the preset intragastric administration amount is calculated according to the body weight of the experimental animals, with each experimental animal being administered 5-10 g / kg each time;

[0009] After acute alcohol pretreatment, LPS was used to induce the experimental animals until depressive-like behaviors appeared, and the constructed alcohol-induced depression model was verified.

[0010] Preferably, in the method for establishing the alcoholic depression animal model, the LPS induction is carried out by intragastric administration of LPS, and each experimental animal is administered 10-15 mg / kg each time according to the body weight of the experimental animals.

[0011] Preferably, in the method for establishing the alcoholic depression animal model, the multiple gavages are 3-5 times, with an interval of 12-18 hours between each gavage.

[0012] Preferably, in the method for establishing the alcoholic depression animal model, LPS is induced in the experimental animals 2-6 hours after the acute alcohol gavage pretreatment.

[0013] Preferably, in the method for establishing the alcoholic depression animal model, the acute alcohol is administered intragastrically three times, at a rate of 5 g / kg per animal according to the body weight of the experimental animals, with an interval of 12 hours between each administration.

[0014] Preferably, in the method for establishing the alcoholic depression animal model, the LPS induction is a single induction, and the induction is performed with an intragastric dose of 10 mg / kg per animal according to the body weight of the experimental animals.

[0015] According to another aspect of the present invention, a depression model constructed by the method for establishing an alcohol-induced depression model of the present invention is provided.

[0016] According to another aspect of the present invention, there is also provided a use of a depression model obtained by the method for establishing an alcoholic depression model of the present invention in screening anti-alcoholic depression drugs.

[0017] In general, compared with the prior art, the above technical solution conceived by the present invention can achieve the following beneficial effects due to the combined induction of acute alcohol and LPS:

[0018] According to the body weight of the experimental animals, each experimental animal is gavaged with 5-10g / kg of acute alcohol each time. The experimental animals are first treated with acute alcohol by gavage multiple times, and then induced with LPS. An alcoholic depression model can be induced within 2-4 days, and the mice have significant depressive behavior. The chronic alcohol-induced depression model requires more than 6 months. Compared with the existing chronic alcohol-induced alcoholic depression model, this method can greatly shorten the experimental modeling time, and LPS can be induced by gavage. It can be seen that the alcoholic depression model constructed by this method has a short modeling cycle, significant depressive-like behavior in mice, and can better simulate the depressive-like behavior of alcoholics. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is a schematic diagram of the process of building the alcohol-depression model;

[0020] Figure 2 It is the verification result of the tail suspension test;

[0021] Figure 3 It is the verification result of the distance traveled in the entire area of ​​the open field experiment;

[0022] Figure 4 It is the time verification result of the peripheral area of ​​the open field experiment;

[0023] Figure 5 This is the verification result of the forced swimming experiment. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0025] People with depression often show low mood, and drinking can help distract from persistent sadness. Although alcohol may temporarily relieve some symptoms of depression, it can easily lead to alcoholism and even develop into alcohol dependence. Depression can easily lead to alcoholism, and alcoholism can aggravate depression, thus forming a vicious circle and ultimately aggravating depression. Studies have also found that the neurophysiological and metabolic changes caused by alcoholism are related to the pathogenesis of depression, but some studies have shown that acute alcohol treatment is difficult to induce depressive behavior in mice. Although depression can put a person at greater risk of developing alcohol dependence, it is more common for alcohol dependence to develop into depression. Currently, patients with alcoholic depression are also attracting more attention.

[0026] Existing studies have found that chronic alcohol treatment can induce depressive behavior in mice, but the induction period is long, usually requiring more than 6 months, and the alcoholic depression model is of a single type, which is not conducive to a comprehensive understanding of the pathogenesis between alcohol and depression. Therefore, studying a rapid induction of alcoholic depression model will help promote the study of the potential mechanism of alcohol and depression or anti-alcoholic depression drugs.

[0027] The commonly used classic depression model is the LPS acute stress model, which uses LPS induction to induce depressive behavior in model animals. LPS induction generally uses LPS injection to induce depression models, such as intraperitoneal injection and intravenous injection, and its induction period is usually 4-7 days. Although the induction period is short, this LPS injection induction method does not match the actual situation of alcoholic depression. In addition, a single injection of LPS induces depressive-like behavior for a short period of time and milder depressive symptoms, while alcoholism often leads to severe depression. Therefore, the existing classic LPS induction model is difficult to simulate the depressive-like behavior of alcoholics in a targeted manner.

[0028] However, our experiments found that the combination of acute alcohol and LPS can stably induce depressive-like behavior in mice. By first administering acute alcohol pretreatment by gavage and then inducing it with LPS, the mice showed depressive behavior 2-4 days after induction, and the depressive behavior of the mice was significant. We further found that LPS can be induced by gavage, and this method of inducing depressive behavior by gavage of LPS is more in line with the actual situation of depressive-like behavior in alcoholics. This induction method can better simulate the depressive behavior of patients with alcoholic depression.

[0029] The present invention provides a method for establishing an alcoholic depression animal model, which comprises the following steps:

[0030] (1) Acute alcohol pretreatment: The experimental animals are pretreated with a preset amount of acute alcohol by gavage multiple times. The preset amount of acute alcohol is calculated based on the weight of the experimental animals, with each gavage being 5-10 g / kg. The multiple gavages are preferably 3-5 times, with an interval of 12-18 hours between each gavage, to simulate the drinking behavior of alcoholics.

[0031] (2) LPS-induced depression model: 2-6 h after acute alcohol pretreatment in step (1), LPS is used to induce depression-like behavior in experimental animals, and the constructed alcoholic depression model is verified; the constructed alcoholic depression model can be verified by behavioral tests, such as open field test, tail suspension test and forced swimming test.

[0032] Preferably, a single intragastric administration of LPS is used for induction, with each animal being administered 10-15 mg / kg according to the weight of the experimental animals.

[0033] Acute alcohol gavage pretreatment combined with LPS induction can induce depressive-like behavior in mice within 2-3 days. Compared with the induction period of chronic alcohol, this method can greatly shorten the experimental modeling time. In addition, in the classic depression modeling method, LPS is induced by intravenous or intraperitoneal injection, while the depression model can be induced by gavage of LPS after multiple acute alcohol gavage treatments. This method of induction by gavage of LPS is more in line with the actual situation of alcoholic depression, and can better simulate the depressive-like behavior caused by alcoholism in the human body, which is conducive to exploring the potential mechanism of the relationship between alcohol and depression. In addition, the depressive behavior of mice is obvious and the model effect is good.

[0034] In addition, the present invention also provides a depression model constructed by the method for establishing an alcohol-induced depression model according to the present invention.

[0035] In addition, the present invention also provides an application of a model obtained by the method for establishing an alcoholic depression model as described in the present invention in screening anti-alcoholic depression drugs.

[0036] The following are examples:

[0037] Example 1 Alcohol + LPS induced alcoholic depression model (mALPS model)

[0038] The flowchart of constructing the alcohol depression model using this method is as follows: Figure 1 As shown, the specific steps are as follows:

[0039] (1) Alcohol pretreatment: Each mouse was given three acute alcohol gavages. The dose was 5 g / kg per mouse according to the mouse body weight, and the interval between each gavage was 12 hours. Physiological saline was used as the blank control group.

[0040] (2) LPS gavage-induced depression model: 2 hours after the third acute alcohol gavage, mice were gavaged with LPS once, with the gavage dose of 10 mg / kg per mouse according to the weight of the mouse; behavioral tests were performed on the mice on days 3-4. The results are shown in the table. Figure 2-5 Middle mALPS group.

[0041] Comparative Example 1 Alcohol-induced depression model (EtOH model)

[0042] Experimental method: Each mouse was given three acute alcohol gavages (5g / kg / dose), and the control group was given the same amount of saline, with an interval of 12 hours between each gavage; 2 hours after the third acute alcohol gavage, the control group and the alcohol group were given the same amount of saline, with the saline gavage volume being 10mg / kg; the behavior of the mice was tested on the 3rd and 4th days, and the results are shown in Figure 2-5 EtOH group.

[0043] Comparative Example 2: Depression Model Induced by Oral Gavage with LPS

[0044] Experimental method: Each mouse was gavaged with saline three times, with a gavage volume of 5g / kg / dose. 2 hours after the third gavage with saline, a single gavage with LPS was performed, with a gavage volume of 10mg / kg for each mouse according to the mouse weight. The control group was gavaged with the same amount of saline. The behavior of the mice was tested on the 3rd and 4th day. The results are shown in Figure 2-5 LPS group.

[0045] Above Figure 2-5 ,in Figure 2 This is the verification result of the tail suspension test. Figure 3 The distance traveled in the entire area of ​​the open field experiment is verified. Figure 4 This is the time verification result of the peripheral area of ​​the open field experiment. Figure 5 Verification results of the forced swimming experiment.

[0046] Depend on Figure 2-5 It can be seen that compared with the control group, the mice in the acute alcohol gavage alone and the LPS gavage group did not show depressive-like behavior, while the mice in the mALS group (acute alcohol + LPS) showed obvious depressive-like behavior, indicating that the acute alcohol + LPS-induced alcohol depression model in mice was successfully established, while the depression models induced by acute alcohol gavage or LPS gavage alone failed.

[0047] In summary, the mining field experiment, tail suspension experiment, and forced swimming experiment all show that this method can be used to construct an alcoholic depression model, and the mice have significant depressive-like behaviors and the model effect is good.

[0048] It will be easily understood by those skilled in the art that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for establishing an alcohol-induced depression animal model, characterized in that: The following steps are involved: Select experimental animals that meet the experimental requirements; The experimental animals are pretreated by multiple intragastric administration of a preset amount of acute alcohol; the preset intragastric administration amount is calculated based on the weight of the experimental animals, with each experimental animal being administered 5-10 g / kg each time, and administered 3-5 times, with an interval of 12-18 hours between each administration; After 2-6 hours of acute alcohol pretreatment, LPS was administered intragastrically to induce the experimental animals until depression-like behaviors appeared, and the constructed alcohol-induced depression model was verified; The LPS induction by intragastric administration is calculated based on the body weight of the experimental animals, with each experimental animal being administered 10-15 mg / kg each time.

2. The method for establishing an alcohol-induced depression animal model according to claim 1, characterized in that: The acute alcohol was administered intragastrically 3 times at a rate of 5 g / kg / time according to the body weight of the experimental animals, with an interval of 12 hours between each administration.

3. The method for establishing an alcohol-induced depression animal model according to claim 2, characterized in that: The LPS induction was a single induction, and the induction was performed with an intragastric dose of 10 mg / kg per animal according to the body weight of the experimental animals.

4. A depression model constructed by the method for establishing an alcohol-induced depression animal model according to any one of claims 1 to 3.

5. Use of a depression model constructed by the method for establishing an alcohol-induced depression animal model according to any one of claims 1 to 3 in screening anti-alcohol depression drugs.

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