Method for establishing depression mouse model

Abstract

The invention relates to the technical field of animal model establishment, in particular to a method for establishing a depression mouse model. Compared with common animal depression modeling modes such as chronic non-stress animal depression modeling modes and constraint stress animal depression modeling modes, the depression model established by the method is short in depression model preparation time and weak in stimulation means, and depression caused by metabolic abnormality due to intestinal microbe abnormality is more specifically simulated. According to the method, intestinal microbial metabolite ethanolamine is externally supplemented for 14 days, so that the mice can have depression behaviors.

Description

Technical Field

[0001] The present invention relates to the technical field of establishing animal models, and particularly relates to a method for establishing a mouse model of depression. Background Art

[0002] As a common mental illness, depression has gradually become a focus of global health concern. Its typical symptoms include persistent low mood, decreased interest, anhedonia, etc. However, due to the high heterogeneity of depression, its pathogenesis covers multiple levels of factors such as biology, psychology, and sociology, making its actual clinical manifestations complex and diverse. Therefore, in order to further understand the mechanism of depression, improving and developing new treatment methods remains an important topic in current psychiatric research.

[0003] Animal models have always been important tools for studying the pathogenesis and drug treatment of depression. Currently, common methods for constructing mouse depression models include the chronic unpredictable stress model (CUMS), forced swimming test (FST), learned helplessness model (LH), etc. However, most traditional models can only simulate some symptoms and mechanisms of depression and cannot comprehensively and accurately reflect the complex clinical manifestations and pathological processes of depression. Therefore, considering the diverse causes and pathogenesis of depression, there is an urgent need to develop a new animal model to better simulate real depression symptoms. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for establishing a mouse model of depression.

[0005] The technical problems solved by the present invention are achieved by adopting the following technical solutions:

[0006] In the first aspect of the present invention, a method for establishing a mouse model of depression is provided, including: mice are administered ethanolamine at a dose of 100 - 200 mg / kg for 7 - 20 days, and during the administration period, the mice drink drinking water containing 2000 - 6000 μM of ethanolamine.

[0007] Based on the deficiencies of existing mouse depression models and the possible relationship between gut microbiota, ethanolamine, and depression, a new method for constructing a mouse depression model induced by ethanolamine is thus constructed, hoping to more accurately simulate the pathophysiological characteristics of human depression and provide a more effective tool for the research of depression.

[0008] Compared with the commonly used restraint stress modeling method, the mouse depression model established by the present invention shows more obvious performance in the novelty suppressed feeding and forced swimming tests after ethanolamine treatment.

[0009] In one or more embodiments of the present invention, the mice are male mice aged 6 - 8 weeks.

[0010] In one or more embodiments of the present invention, the dosage of ethanolamine is 140-180 mg / kg, and the content of ethanolamine in the drinking water is 3000-5000 μM.

[0011] In one or more embodiments of the present invention, the dosage of ethanolamine is 150-170 mg / kg, and the content of ethanolamine in the drinking water is 3500-4500 μM.

[0012] In one or more embodiments of the present invention, the dosage of ethanolamine is 150-170 mg / kg, and the content of ethanolamine in the drinking water is 3500-4500 μM.

[0013] In one or more embodiments of the present invention, the dosage of ethanolamine is 155-165 mg / kg, and the content of ethanolamine in the drinking water is 3800-4200 μM.

[0014] In one or more embodiments of the present invention, the intake of drinking water is 3-8 ml.

[0015] In one or more embodiments of the present invention, the intake of drinking water is 4-6 ml.

[0016] In one or more embodiments of the present invention, the administration time of ethanolamine is 10-18 days.

[0017] In one or more embodiments of the present invention, the administration time of ethanolamine is 12-16 days

[0018] The present invention includes the following beneficial effects: The depression model established in the present invention, compared with the commonly used animal depression modeling methods such as chronic unstressed and restraint stress, has a short time for preparing the depression model, weak stimulation means, and more specifically simulates the depression caused by abnormal intestinal microbiota leading to abnormal metabolism. By externally supplementing the intestinal microbiota metabolite ethanolamine for 14 days, depressive behavior can occur in mice by this method. Description of the Drawings

[0019] Figure 1 Is the feeding latency of animals G1 and G2 in the novelty suppressed feeding test.

[0020] Figure 2 Is the struggling time of animals G1 and G2 in the forced swimming test.

[0021] Figure 3 Is the weight change of animals G1 and G2 during the experiment.

[0022] Figure 4 Is the time in the center of the open field test (OFT) for the animals.

[0023] Figure 5 is the feeding latency of animals in the novelty suppressed feeding test.

[0024] Figure 6 is the self-care behavior of animals in the splash test (ST).

[0025] Figure 7 is the immobility time of animals in the forced swimming test (FST).

[0026] Figure 8 is the device diagram of the conventional restraint stress group.

[0027] Unless there is a contrary statement, the terms used in the specification and claims have the following meanings.

[0028] As used in the present invention, the term "prevention" refers to preventing the occurrence of a disease and / or preventing the recurrence of a disease. Detailed Description of the Invention

[0029] The specification of the present invention has described the specific implementation manners in detail. Those skilled in the art should recognize that the following implementation manners are exemplary and should not be construed as limitations on the present invention. For those skilled in the art, without departing from the principle of the present invention, by making several improvements and modifications to the present invention, the technical solutions obtained by these improvements and modifications also fall within the protection scope of the claims of the present invention. The beneficial effects of the present invention are specifically described below through examples.

[0030] Example 1

[0031] Establishing a mouse depression model with ethanolamine

[0032] 1. Mouse depression model

[0033] 1.1 Establishing a depression model with ethanolamine (G1)

[0034] Select male C57 mice aged 6 - 8 weeks. Continuously intragastrically administer ethanolamine to the mice for 14 days. According to the body weight of the experimental animals, calculate it as 160 mg / kg per gavage per mouse each time. At the same time, let the mice drink purified water containing 4000 μM ethanolamine in daily life (about 5 ml is ingested).

[0035] 1.2 Conventional restraint stress group (G2)

[0036] Each C57 mouse in this group receives restraint stress with different durations of 4 - 8 hours and indefinite restraint times every day. The specific process is to place the mouse in a fixator, making the animal's torso and head and neck unable to twist, but ensuring its normal breathing. During this period, it does not contact food and drinking water. Specifically, see the following Figure 8 .

[0037] 1.3 Normal control group (G3)

[0038] Each C57 mouse in this group was raised completely normally in the cage without any treatment or operation.

[0039] 2. Detection of depressive behavior

[0040] 2.1 Novelty suppressed feeding test (NSFT)

[0041] After the mice were fasted for 48 hours in advance, they were placed in a square open field (50×50 cm). A small amount of feed (placed on filter paper) was placed in the middle of the open field. During the experiment, the mouse was placed in the corner of the open field with its head facing the corner and the timing started, and the timing stopped when it started to eat. The entire test time did not exceed 5 min. After the test, the mouse was immediately returned to the original cage. After each mouse was tested, the feed was replaced and the open field was wiped with alcohol to avoid the smell information of the mouse. It is used to simulate anhedonia and reduced interest in depression.

[0042] Figure 1 is the feeding latency of animals in G1 and G2 in the novelty suppressed feeding test. Etn is the ethanolamine treatment group, and Model is the conventional restraint stress group. It can be seen that ethanolamine treatment can lead to a longer feeding latency. The novelty suppressed feeding test reflects the degree of depression and anxiety level of rodent feeding, because there is a contradictory competition between the animal's desire to eat and the fear of the novel space, which enables this experiment to effectively reflect the anhedonia of clinically depressed patients. The longer the feeding latency time, the lower the exploratory ability of the mouse in the novel space and the lower the desire for things.

[0043] Figure 2 is the feeding latency of animals in G1 and G3 in the novelty suppressed feeding test. Etn is the ethanolamine treatment group, and Control is the normal control group. It can be seen that ethanolamine treatment can lead to a longer feeding latency.

[0044] Figure 1 and Figure 2 refers to the comparison between the ethanolamine model and the conventional restraint stress model. The normal restraint model should be 3 weeks. It can be seen from this figure that the ethological results at the end of the 14-day modeling can show that the depressive-like behavior of the mice in the ethanolamine modeling group is more obvious; therefore, it shows that the time for preparing the depressive model by this method is short. 2.2 Splash test (ST)

[0045] After spraying a 10% sucrose solution on the dorsal hair of the animal and placing it in its cage, the frequency and duration of the animal's grooming were recorded within 5 minutes. The viscosity of the sucrose solution mimics the dirt on the fur of rodents and triggers grooming by licking or scratching. The time delay between spraying and the start of grooming and the decrease in grooming frequency are considered to be a reduction in motivation and self-care behavior associated with a depressive-like phenotype.

[0046] Figure 3 Self-care behavior of the animal in the splash test (ST). Etn is the ethanolamine treatment group, and Model is the conventional restraint stress group. It can be seen that ethanolamine treatment can lead to a decrease in self-care behavior (grooming time). Figure 3 It can be seen that compared with restraint stress, the body weight of the ethanolamine model animals decreased insignificantly, indicating that this method has a weak stimulating means.

[0047] 2.3 Forced swim test (FST)

[0048] The mouse was placed in a transparent glass cylinder filled with water (40 cm high and 20 cm in diameter), the water level was 30 cm high, the water temperature was maintained at 25 °C, and the time was recorded for 6 min. The cumulative immobility time, swimming time, and struggling time within the last 4 min were recorded and analyzed to evaluate the depressive "behavioral despair" symptoms.

[0049] Figure 4 Struggling time of the animal in the forced swim test. Etn is the ethanolamine treatment group, and Model is the conventional restraint stress group. It can be seen that ethanolamine treatment can lead to less struggling.

[0050] Figure 5 Immobility time of the animal in the forced swim (FST) test. Etn is the ethanolamine treatment group, and Control is the normal control group. It can be seen that ethanolamine treatment can lead to an increase in immobility time.

[0051] 2.4 Open field test (OFT)

[0052] The mouse was placed in a clean test box (50×50 cm), and the total movement distance, standing times, and the number of entries into the central area of the mouse were observed and recorded for 5 min (the central area refers to the area in the middle of the test box that accounts for 50% of the total area). Mice like to stick close to the wall because of thigmotaxis, that is, they move more around the perimeter and in the corners. Depressed mice tend to stay in the central area, i.e., the open area, of the open field for a shorter time. It is used to simulate the autonomous activity ability and exploratory behavior of depression.

[0053] Figure 6Time in center in the open field test (OFT) for animals. Etn is the ethanolamine treatment group, and Control is the normal control group. As can be seen from the figure, ethanolamine treatment can lead to a decrease in time in center.

[0054] The splash test (ST), novelty suppressed feeding (NSFT), forced swimming test (FST), and open field test (OFT) were used to evaluate states such as reduced self-care for depression, anhedonia, and despair behavior, respectively. It was found that compared with normal control mice (Control), mice treated with ethanolamine (Etn) showed depressive-like behaviors such as reduced grooming time in the splash test, increased feeding latency in the novelty suppressed feeding test, increased immobility time in the forced swimming test, and decreased time in center in the open field test. The establishment of this depression model was evaluated as successful.

[0055] The description of specific implementation manners in the specification of the present invention has been detailed. Those skilled in the art should recognize that the above implementation manners are exemplary and should not be construed as limitations on the present invention. For those skilled in the art, without departing from the principle of the present invention, by making several improvements and modifications to the present invention, the technical solutions obtained from these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A method for establishing a mouse model of depression, characterized in that, Including: The mice are administered ethanolamine at a dose of 100 - 200 mg / kg for 7 - 20 days. During the administration period, the mice drink drinking water containing 2000 - 6000 μM of ethanolamine.

2. The method according to claim 1, characterized in that, The mice are male mice at 6 - 8 weeks old.

3. The method according to claim 1, characterized in that, The dose of the ethanolamine is 140 - 180 mg / kg, and the content of ethanolamine in the drinking water is 3000 - 5000 μM.

4. The method according to claim 3, characterized in that, The dose of the ethanolamine is 150 - 170 mg / kg, and the content of ethanolamine in the drinking water is 3500 - 4500 μM.

5. The method according to claim 3, characterized in that, The dose of the ethanolamine is 150 - 170 mg / kg, and the content of ethanolamine in the drinking water is 3500 - 4500 μM.

6. The method according to claim 3, characterized in that The dose of the ethanolamine is 155 - 165 mg / kg, and the content of ethanolamine in the drinking water is 3800 - 4200 μM.

7. The method according to claim 1, characterized in that The intake of the drinking water is 3 - 8 ml.

8. The method according to claim 7, wherein The intake of the drinking water is 4 - 6 ml.

9. The method according to claim 1, characterized in that, The administration time of the ethanolamine is 10 - 18 days.

10. The method according to claim 9, characterized in that, The administration time of the ethanolamine is 12 - 16 days.

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