A method, device and application for constructing a bipolar disorder experimental animal model

Abstract

The application relates to the field of medical biology, and discloses a construction method, device and application of a bipolar disorder experimental animal model, the construction method being as follows: healthy experimental animals are selected for pre-molding environmental adaptability feeding, then sleep deprivation stimulation lasting for 3-5 days is carried out, wherein the experimental animals experience continuous 15-18 hours of sleep deprivation and 6-9 hours of rest every day, and the experimental animals are free to eat and drink; after the sleep deprivation stimulation is finished, the experimental animals are subjected to behavior test evaluation to evaluate the surface validity of the disease model. The construction method is simple to operate and easy to repeat, can better simulate the symptoms of clinical bipolar disorder patients, and has a longer disease model maintenance time, so that the application provides a new model paradigm for the research on the pathogenesis of bipolar disorder and the clinical drug screening of bipolar disorder.

Description

Technical Field

[0001] This invention relates to the field of medical biology, specifically to a method for constructing animal models of mental illness, and more specifically to a method, apparatus, and application for constructing an experimental animal model of bipolar disorder. Background Technology

[0002] Bipolar disorder (BD), also known as manic-depressive illness, is a neuropsychiatric disorder characterized by recurrent, alternating, or cyclical episodes of mania (or hypomania) and depression. It typically begins in adolescence or early adulthood and can severely impact a patient's life and social functioning. BD involves both manic or hypomanic episodes (characterized by elevated mood, flight of ideas, and increased volition) and depressive episodes (characterized by depressed mood, slowed thinking, and reduced volition). Its recurrent and chronic nature presents significant challenges to treatment. Current research on BD primarily focuses on the depressive phase, with insufficient research on the manic phase, largely due to the lack of effective animal models. Therefore, developing novel animal models of BD is crucial for understanding the pathogenesis of mania.

[0003] Animal models of bipolar disorder commonly use rats and mice, with mice being the most frequently used. The book *Rodent Models and Behavioral Experiments of Mental Illness*, edited by Cui Donghong and Xu Lin et al. (2024), provides a detailed overview of various BD animal models, primarily including genetic engineering models (Clock gene mutation model, GSK3 overexpression model), drug-induced models (amphetamine-induced model, ouabain-induced model), and environmental stress models (sleep deprivation model). Genetic engineering models significantly impact the normal reproduction and growth of mice, and have a long breeding cycle, resulting in high time and economic costs. Amphetamine, a controlled psychoactive substance, is subject to strict approval procedures and hazardous chemical management regulations for its purchase and use, hence there are few reports in domestic literature. Oouabain, another modeling drug, requires targeted administration, making the modeling process cumbersome. Deviations in stereotactic injection sites can easily lead to model instability, and anesthesia and surgery can also cause mouse mortality, increasing experimental costs. Reduced sleep demand is one of the typical clinical manifestations of manic episodes in BD. Sleep deprivation is also an environmental stressor in BD manic episodes, which can induce BD patients to switch from a depressive state to a manic state. The multi-platform aquatic environment stress model is the most commonly used sleep deprivation model. However, the behavioral changes in mice may not be solely due to sleep deprivation, but may also be influenced by a combination of factors such as aquatic environment stress and limited activity space. It is difficult to accurately attribute them to sleep deprivation itself. When animals are in a multi-platform aquatic environment for a long time, their bodies are in a state of stress, their immunity is reduced, and they are prone to various diseases. This may even lead to infection or drowning in mice. This will not only increase the experimental cost, but also affect the reliability of the experimental results and cannot truly reflect the impact of sleep deprivation on normal physiological functions.

[0004] Chinese utility model patent CN206629696U (authorization announcement date: November 14, 2017) discloses a device for establishing a sleep deprivation model in rodents. The device includes a box-shaped body containing a suitable amount of liquid that does not cause chemical harm to rodents. The depth of the liquid does not exceed the body length of the rodent. At least one support rod is provided above the liquid surface for the rodent to stand on. A top cover has a solid food storage area and a water feeding area for the rodent to eat and drink normally. The top cover is positioned over the opening of the box-shaped body. This device utilizes the rodent's aversion to water, preventing mice or rats from falling asleep by making them stand on the horizontal support rod close to the water surface for an extended period. It is designed for long-term experimental use and allows for strict time control, making it suitable for establishing a sleep deprivation model. However, the device is cumbersome to operate (the water must be changed daily), otherwise mouse excrement and food residue will fall into the water, making it cloudy, deteriorating its quality, and producing an unpleasant odor. Drinking this water can increase the risk of infection and discomfort in the mice. Aquatic stress can also increase confounding factors other than sleep deprivation. These adverse factors not only increase experimental costs but also affect the reliability of experimental results, making them unsuitable for long-term stable modeling.

[0005] Chinese utility model patent CN222193580U discloses a sleep deprivation device for small animals used in bioengineering, including a base with a bottom plate at the top and a rotating shaft at the center of the bottom plate. The lower end of the rotating shaft is connected to a drive motor inside the base. A disturbance rod is mounted on the rotating shaft. A cylinder is mounted on the base, with a top cover at the top and ventilation holes on the top cover. A light is mounted on the bottom surface of the top cover or the side wall of the cylinder. A water feeder and a food feeder are mounted on the outer wall of the cylinder. A touch screen is located on the front of the base. This device uses a disturbance rod instead of a water environment, which can avoid the adverse effects of the water environment on animals. It is relatively simple to operate, has fewer interfering factors, and is suitable for long-term stable modeling. However, this device keeps the animal in a state of sleep disorder due to constant contact with the disturbance rod, preventing the animal from getting rest. Therefore, this type of interference device is often used to construct animal models of depression, but it is not suitable for constructing a mouse model of bipolar disorder.

[0006] Furthermore, there is currently a lack of a unified paradigm for constructing bipolar disorder mouse models through multi-platform aquatic environment sleep deprivation. The book "Rodent Models of Mental Illness and Behavioral Experiments" edited by Cui Donghong and Xu Lin et al. (2024) provides a detailed overview of methods for constructing bipolar disorder models, with a sleep deprivation duration of 72 hours. However, some literature discloses modeling times ranging from 36 to 72 hours. These modeling paradigms are all acute, and after successful BD model construction, the model is only maintained for about 30 minutes, making it impossible to conduct observation experiments on the behavior and physiological indicators of model animals for more than one hour, thus failing to meet the needs for longer-term physiological testing. Summary of the Invention

[0007] To address the problems of short maintenance time, numerous confounding factors, high experimental costs, and poor accuracy of experimental results in existing bipolar disorder mouse models constructed using current paradigms, this invention provides a method, apparatus, and application for constructing a bipolar disorder experimental animal model.

[0008] To achieve the above-mentioned technical objectives, the technical solution adopted by the present invention is as follows:

[0009] In the first aspect, this application provides a method for constructing an experimental animal model of bipolar disorder. Healthy experimental animals are selected for environmental adaptation before modeling, and then subjected to sleep deprivation stimulation for 3 to 5 days, in which they experience 15 to 18 hours of continuous sleep deprivation and 6 to 9 hours of rest each day, with free access to food and water. After the sleep deprivation stimulation ends, behavioral tests are used to evaluate the face validity of the disease model.

[0010] As a preferred technical solution, the sleep deprivation period is 15 to 18 consecutive hours from 16:00 in the afternoon to 10:00 the next morning.

[0011] As a preferred technical solution, sleep deprivation stimulation is performed in a sleep deprivation device, which includes a rotating platform and a chamber. The chamber is placed above the rotating platform, and touch rods and rest rods are arranged radially between the inner walls of the chamber. The touch rods are adjacent to the upper surface of the rotating platform, and the rest rods are spaced above the touch rods. The continuous rotation of the rotating platform and the blocking of the touch rods disturb the sleep of the experimental animals.

[0012] As a preferred technical solution, the rotation speed of the rotating platform is 2 to 30 seconds per revolution.

[0013] As a preferred technical solution, behavioral tests include the open field test and the elevated cross maze test.

[0014] As a preferred technical solution, the open field experiment method is as follows: the experimental animal is placed in a square box made of white plexiglass and allowed to explore the box freely for 5 minutes. During the test, the total distance the experimental animal moves, the frequency of entering the central area, and the time spent in the central area are analyzed by a video tracking system.

[0015] As a preferred technical solution, the elevated cross maze experiment method is as follows: The elevated cross maze experimental device consists of a white plexiglass box with two open arms and two closed arms, arranged in a cross shape, 40-60 cm above the ground; the experimental animal is placed alone in the center of the elevated cross maze device, facing one of the closed arms, and allowed to freely explore the device for 5 minutes; during the test, a video tracking system is used to quantify the total distance the experimental animal moves, the number of times it enters the open arms, and the time it spends on the open arms.

[0016] As a preferred technical solution, the experimental animal is a rodent.

[0017] Secondly, this application provides a sleep deprivation device, including a rotating platform and a housing. The housing is placed above the rotating platform, and a switch is provided on the rotating platform. A touch bar and a rest bar are arranged radially between the inner walls of the housing. The touch bar is adjacent to the upper surface of the rotating platform, and the rest bar is spaced above the touch bar. The continuous rotation of the rotating platform and the blocking of the touch bar disturb the sleep of the experimental animal. The top of the housing is provided with an openable mesh, and a water bottle is provided on the side of the housing. The rotating platform is connected to an intelligent circulation controller, which is provided with a display screen and buttons.

[0018] Secondly, this application provides an application in which an animal model obtained by the method described above is used to evaluate drugs that can alleviate or treat bipolar disorder.

[0019] The beneficial effects of this invention are:

[0020] The method for constructing an experimental animal model of bipolar disorder proposed in this invention is simple to operate and easy to repeat. It can better simulate the symptoms of clinical bipolar disorder patients, and the disease model can be maintained for a long time. It will provide a new model paradigm for the study of the pathogenesis of bipolar disorder and the screening of clinical drugs.

[0021] The method for constructing a bipolar disorder experimental animal model of the present invention provides a novel environmental stress model, enriching the field of bipolar disorder animal models. The inventors have repeatedly tested and optimized parameters such as the ratio of daily modeling time to rest time and the number of days of continuous modeling to ensure the stability and reproducibility of the bipolar disorder mouse model.

[0022] The sleep deprivation device designed in this invention uses a rotating platform instead of a water environment and a disturbance rod. The rotating platform and the touch rod disturb the animal's sleep. At the same time, resting rods, food and water are set above the rotating platform. This can satisfy the experimental animals' need for short rest without preventing them from entering REM sleep, otherwise they would fall due to muscle relaxation. It can also greatly reduce the mortality of mice caused by infection or drowning due to long-term chronic sleep deprivation. The model has a high success rate, small individual differences, good repeatability, and short modeling time. It can better simulate the slow progression of bipolar disorder and is especially suitable for large-scale, high-throughput preclinical animal testing of new drugs, thus accelerating the drug development process.

[0023] The device designed in this invention has a simple structure and low cost, making it suitable for large-scale use and greatly improving scientific research efficiency and saving research costs. Attached Figure Description

[0024] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0025] Figure 1 This is a schematic diagram of the sleep deprivation device in this invention.

[0026] Figure 2 This study analyzed the changes in body weight, open field test, and elevated cross maze test in model mice after sleep deprivation stimulation.

[0027] Figure 3 This is a behavioral analysis of model mice at different time points after the end of sleep deprivation stimulation.

[0028] Figure 4 This study analyzed the open field and elevated cruciate maze experiments in mice after the sleep deprivation stimulus was completed, comparing the control group, treatment group, model group, and treatment / model group.

[0029] Reference numerals: 1-Rotating platform, 2-Switch, 3-Touch lever, 4-Rest lever A, 5-Rest lever B, 6-Box body, 7-Mesh screen, 8-Kettle, 9-Controller, 10-Display screen, 11-Button, 12-Socket. Detailed Implementation

[0030] To make the key technical points, technical solutions, and beneficial effects of this invention clearer, the invention will be described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the embodiments are only for explaining the invention and are not intended to limit the scope of the invention.

[0031] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Other materials and reagents used, unless otherwise specified, are commercially available. All animal experiments conducted have been approved by the Laboratory Animal Ethics Committee of West China Hospital, Sichuan University (Ethics Filing No.: 20230504005).

[0032] A method for constructing a bipolar disorder experimental animal model includes the following steps:

[0033] (1) Selection of experimental animals: Select several experimental animals that match the experimental needs in terms of strain, sex, age, weight, etc., and randomly group them. The experimental animals are rodents, such as mice and rats.

[0034] (2) Adaptation before modeling: The experimental animals were first fed to adapt to the modeling environment for 7 days. The experimental animals were allowed to eat and drink freely. The experimental personnel observed the condition of the experimental animals every other day, supplemented their food and water, and recorded the weight information of the experimental animals before the modeling began.

[0035] (3) Sleep deprivation stimulation: At the beginning of the modeling, the parameters of the sleep deprivation device are first set, including the device's start time, shutdown time, number of cycles, etc. In one embodiment of the present invention, the parameters are that the rotating platform of the sleep deprivation device runs continuously for 3 to 5 days, 15 to 18 hours a day, with the running time period from 16:00 in the afternoon to 10:00 the next morning, followed by a rest period of 6 to 9 hours, and the rotation speed of the platform is 2 to 30 seconds / revolution. Then, the experimental animals are placed in the box of the sleep deprivation device in advance, covered with a mesh, and given food and drinking water. The experimental personnel observe the condition of the experimental animals every other day, and replenish food and water. Preferably, the experimental animals are deprived of sleep for 16 hours and rest for 8 hours a day, and the modeling lasts for 4 days.

[0036] (4) Behavioral evaluation of experimental animals in bipolar disorder model: The face validity of the disease model was evaluated using behavioral tests, including the open field test and the elevated cross maze test.

[0037] (5) Validation of the experimental animal model of bipolar disorder: Valproate (a first-line clinical drug for bipolar disorder) was used to evaluate the predictive validity of the disease model. The dosage of valproate was 200 mg / kg, administered by intraperitoneal injection once daily for 7 days. After the valproate treatment was completed, behavioral tests were used to evaluate the therapeutic effect of the drug.

[0038] The above-mentioned method for constructing an experimental animal model of bipolar disorder uses a sleep deprivation device, such as... Figure 1 As shown: The device includes a rotating platform 1, which is equipped with a switch 2 for controlling the power-on of the device and the rotation direction of the rotating platform; it also includes a cylindrical housing 6, which covers the rotating platform 1. A touch rod 3 is radially arranged on the lower part of the inner wall of the housing 6, adjacent to the upper surface of the rotating platform 1, with a distance of 5-10 mm between the touch rod 3 and the upper surface of the rotating platform 1. Resting rods A4 and B5 are also radially staggered on the inner wall of the housing 6, with resting rod A4 located 3-5 cm above the rotating platform 1 and resting rod B5 located 7-10 cm above the rotating platform 1. The top of the housing 6 has an openable perforated mesh 7 with a spacing of 1 cm, serving both ventilation and feeding trough functions. A water bottle 8 is located on the side of the housing 6. The rotating platform 1 is connected to an intelligent circulation controller 9, which consists of a display screen 10, buttons 11, and sockets 12. Preferably, the rotating platform 1 has a diameter of 14 cm, the box 6 is made of transparent material, and the inner diameter and height of the box 6 are 15 cm. The dimensions of the box and the rotating platform can also be customized according to the body size of mice and rats to be used for sleep deprivation stimulation modeling in experimental animals of different body sizes.

[0039] Example 1

[0040] This embodiment 1 provides a method for constructing a bipolar disorder mouse model and behavioral analysis.

[0041] (1) Selection of mice: SPF-grade mice without any treatment were provided by Chengdu Yaokang Biotechnology Co., Ltd. The strain was C57BL / 6J, the sex was male, the age was 7 weeks, and the weight was 21-23 grams. The mice were kept in a clean environment with the lights on from 7:00 to 19:00.

[0042] (2) Adaptation before modeling: The mice were first fed to adapt to the modeling environment for 7 days. The mice had free access to food and water. The researchers observed the mice's condition every other day and supplemented their food and water. Before the modeling began, the mice's weight information was recorded. The mice were randomly divided into two groups: a control group (n=9) and a model group (n=9).

[0043] (3) Sleep deprivation stimulation: At the beginning of the modeling process, the sleep deprivation device (e.g., sleep deprivation device) should be set up first. Figure 1The parameters of the device were set, and the device was turned on from 18:00 to 10:00 the next morning every day (running continuously for 16 hours). That is, the rotating platform of the sleep deprivation device ran continuously for 16 hours, and the rotation speed of the platform was 30 seconds / revolution. Then, the mice were placed in the sleep deprivation device box in advance (before 18:00), the mesh was covered, and food and drinking water were added. The researchers observed the mice's condition every other day, and replenished food and water. The mice were deprived of sleep for 16 hours and rested for 8 hours every day. The model lasted for 4 days.

[0044] (4) Behavioral evaluation of the bipolar disorder model mice: After modeling, an open field test was conducted. The open field test setup consisted of a box made of white plexiglass (40 cm × 40 cm × 35 cm). Each mouse was placed in a corner of the test area and allowed to freely explore the setup for 5 minutes. After each test, the open field test setup was cleaned with a 75% alcohol solution to remove any odor traces. During the 5-minute test, the total distance the mice moved, the frequency of entering the central area, and the time spent in the central area were analyzed using a video tracking system (Nordas, Netherlands). Then, an elevated cross maze test was conducted. The elevated cross maze setup consisted of a white plexiglass box with two open arms (30 cm long × 7 cm wide) and two closed arms (30 cm long × 7 cm wide × 16 cm high). The two closed arms and two open arms were arranged in a cross shape, 60 cm above the ground. A camera was fixed above the elevated cross maze to record the mice's activities. Each mouse was placed individually in the center of an elevated cross maze, facing one of the closed arms, and allowed to freely explore the device for 5 minutes. After each experiment, the device was cleaned with a 75% alcohol solution to remove any odor traces. During the 5-minute test, a video tracking system was used to quantify the total distance the mouse moved, the number of times it entered an arm, and the time it spent on the open arm.

[0045] Control group: Mice were placed in cages without any treatment and directly subjected to open field test and elevated cross maze test.

[0046] Model group: Mice were treated with the above sleep deprivation stimulus (3), and after the model was completed, open field test and elevated cross maze test were performed.

[0047] Experimental results are as follows Figure 2 As shown, the results indicated that sleep deprivation stimulation significantly reduced the body weight of mice (P<0.01) and significantly increased the distance of movement, the time and frequency of dwell in the central area in the open field (P<0.01), as well as the distance of movement, the time and frequency of dwell in the open arm in the elevated cross maze (P<0.01). The experimental results confirmed that the model mice exhibited manic-like behavior and could simulate the symptoms of bipolar disorder. The model has passed the face validity assessment.

[0048] Example 2

[0049] This embodiment 2 provides an analysis of the maintenance time of a bipolar disorder mouse model.

[0050] (1) Selection of mice: SPF-grade mice without any treatment were provided by Chengdu Yaokang Biotechnology Co., Ltd. The strain was C57BL / 6J, the sex was male, the age was 7 weeks, and the weight was 21-23 grams. The mice were kept in a clean environment with the lights on from 7:00 to 19:00.

[0051] (2) Adaptation before modeling: The mice were first fed to adapt to the modeling environment for 7 days. The mice had free access to food and water. The researchers observed the mice's condition every other day and supplemented their food and water. The mice's weight information was recorded before the modeling began. The mice were randomly divided into 5 groups: control group (11 mice), model group-1h (9 mice), model group-4h (7 mice), model group-12h (7 mice) and model group-24h (10 mice).

[0052] (3) Sleep deprivation stimulation: At the beginning of the modeling process, the sleep deprivation device (e.g., sleep deprivation device) should be set up first. Figure 1 The parameters of the device were set, and the device was turned on from 18:00 to 10:00 the next morning every day (running continuously for 16 hours). That is, the rotating platform of the sleep deprivation device ran continuously for 16 hours, and the rotation speed of the platform was 30 seconds / revolution. Then, the mice were placed in the box of the sleep deprivation device in advance (before 18:00), the mesh was covered, and food and drinking water were added. The researchers observed the mice's condition every other day, and replenished food and water. The mice were deprived of sleep for 16 hours and rested for 8 hours every day. The model lasted for 4 days.

[0053] (4) Behavioral evaluation of the bipolar disorder model mice: After modeling, an open field test was conducted. The open field test apparatus consisted of a box made of white plexiglass (40 cm × 40 cm × 35 cm). Each mouse was placed in a corner of the test area and allowed to freely explore the apparatus for 5 minutes. After each test, the open field test apparatus was cleaned with a 75% alcohol solution to remove any odor traces. During the 5-minute test, parameters such as the total distance the mice moved were analyzed using a video tracking system (Nordas, Netherlands).

[0054] Control group: Mice were placed in cages without any treatment and directly subjected to open field experiments.

[0055] Model group -1h: Mice were treated according to the above steps (3). One hour after the modeling was completed, an open field experiment was conducted.

[0056] Model group -4h: Mice were treated according to the above steps (3). Four hours after the modeling was completed, an open field experiment was conducted.

[0057] Model group -12h: Mice were treated according to the above steps (3). Twelve hours after the modeling was completed, an open field experiment was conducted.

[0058] Model group -24h: Mice were treated according to the above steps (3). 24 hours after the modeling was completed, an open field experiment was conducted.

[0059] Experimental results are as follows Figure 3 As shown, the results indicated that sleep deprivation stimulation significantly increased the activity level of mice (P<0.01), resulting in manic-like behavior. The model could be maintained for more than 4 hours, which met the requirements for long-term observation of the behavior and physiological indicators of the model animals.

[0060] Example 3

[0061] This embodiment 3 provides a method for constructing a bipolar disorder mouse model and an evaluation of the model's predictive validity.

[0062] (1) Selection of mice: SPF-grade mice without any treatment were provided by Chengdu Yaokang Biotechnology Co., Ltd. The strain was C57BL / 6J, the sex was male, the age was 7 weeks, and the weight was 21-23 grams. The mice were kept in a clean environment with the lights on from 7:00 to 19:00.

[0063] (2) Pre-modeling adaptation: The mice were first fed to adapt to the modeling environment for 7 days. The mice had free access to food and water. The researchers observed the mice's condition every other day and supplemented their food and water. The mice were randomly divided into 4 groups: control group (11 mice), treatment group (10 mice), model group (12 mice) and treatment / model group (12 mice).

[0064] (3) Sleep deprivation stimulation: At the beginning of the modeling process, the sleep deprivation device (e.g., sleep deprivation device) should be set up first. Figure 1 The parameters of the device were set, and the device was turned on from 18:00 to 10:00 the next morning every day (running continuously for 16 hours). That is, the rotating platform of the sleep deprivation device ran continuously for 16 hours, and the rotation speed of the platform was 30 seconds / revolution. Then, the mice were placed in the sleep deprivation device box in advance (before 18:00), the mesh was covered, and food and drinking water were added. The researchers observed the mice's condition every other day, and replenished food and water. The mice were deprived of sleep for 16 hours and rested for 8 hours every day. The model lasted for 4 days.

[0065] (4) Behavioral evaluation of the bipolar disorder model mice: After modeling, an open field test was conducted. The open field test setup consisted of a box made of white plexiglass (40 cm × 40 cm × 35 cm). Each mouse was placed in a corner of the test area and allowed to freely explore the setup for 5 minutes. After each test, the open field test setup was cleaned with a 75% alcohol solution to remove any odor traces. During the 5-minute test, the total distance the mice moved, the frequency of entering the central area, and the time spent in the central area were analyzed using a video tracking system (Nordas, Netherlands). Then, an elevated cross maze test was conducted. The elevated cross maze setup consisted of a white plexiglass box with two open arms (30 cm long × 7 cm wide) and two closed arms (30 cm long × 7 cm wide × 16 cm high). The two closed arms and two open arms were arranged in a cross shape, 60 cm above the ground. A camera was fixed above the elevated cross maze to record the mice's activities. Each mouse was placed individually in the center of an elevated cross maze, facing one of the closed arms, and allowed to freely explore the device for 5 minutes. After each experiment, the device was cleaned with a 75% alcohol solution to remove any odor traces. During the 5-minute test, a video tracking system was used to quantify the total distance the mouse moved, the number of times it entered an arm, and the time it spent on the open arm.

[0066] Control group: Mice were placed in cages and treated in parallel. Then, they underwent open field and elevated cross maze tests.

[0067] Treatment group: Mice were placed in cages and the predictive validity of the disease model was evaluated using valproate (a first-line clinical drug for bipolar disorder). The dosage of valproate was 200 mg / kg, administered via intraperitoneal injection once daily for 7 days. After the valproate treatment was completed, the open field test and the elevated cruciate maze test were performed.

[0068] Model group: Mice were treated according to the above steps (3), and after the modeling was completed, open field test and elevated cross maze test were performed.

[0069] Treatment / Model Group: Mice were treated according to step (3) above, and simultaneously treated with valproate. After the treatment, open field test and elevated cross maze test were performed.

[0070] Experimental results are as follows Figure 4As shown, the results indicated that sleep deprivation stimulation significantly increased the distance of movement, the time and frequency of dwell in the central area in the open field of mice (P<0.01), and the distance of movement, the time and frequency of dwell in the open arm in the elevated cruciate maze (P<0.05). Valproate (a first-line clinical drug for bipolar disorder) treatment significantly reversed the above changes in the model mice (P<0.001). The experimental results have confirmed that the novel bipolar disorder mouse model proposed in this invention passed the predictive validity assessment.

[0071] The above results demonstrate that the method for constructing a bipolar disorder mouse model provided in this embodiment of the invention can quickly obtain an ideal mouse model.

[0072] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, or improvements made within the principles and guidelines of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for constructing an experimental animal model of bipolar disorder, characterized in that: Healthy laboratory animals were selected and subjected to environmental adaptation before modeling. Then, they were subjected to sleep deprivation stimulation for 3 to 5 days, during which they experienced 15 to 18 hours of continuous sleep deprivation and 6 to 9 hours of rest each day, with free access to food and water. The sleep deprivation period was 15 to 18 hours from 16:00 in the afternoon to 10:00 the next morning. After the sleep deprivation stimulation ended, the face validity of the disease model was evaluated by behavioral tests on the laboratory animals. Sleep deprivation stimulation is conducted in a sleep deprivation device, which includes a rotating platform and a chamber. The chamber is positioned above the rotating platform. Touch rods are radially arranged on the lower part of the inner wall of the chamber, adjacent to the upper surface of the rotating platform, at a distance of 5-10 mm. Resting rods A and B are also radially staggered on the inner wall of the chamber. Resting rod A is located 3-5 cm above the rotating platform, and resting rod B is located 7-10 cm above the rotating platform. The continuous rotation of the platform and the blocking effect of the touch rods disrupt the sleep of the experimental animals. The rotation speed of the platform is 2-30 seconds per revolution. The resting rods above the rotating platform provide brief rest for the experimental animals without preventing them from entering REM sleep, which would otherwise cause them to fall due to muscle relaxation.

2. The method for constructing a bipolar disorder experimental animal model according to claim 1, characterized in that: Behavioral tests include the open field test and the elevated cross maze test.

3. The method for constructing a bipolar disorder experimental animal model according to claim 2, characterized in that: The open field experiment method is as follows: the experimental animal is placed in a square box made of white plexiglass and allowed to explore the box freely for 5 minutes. During the test, the total distance the experimental animal moves, the frequency of entering the central area, and the time spent in the central area are analyzed by a video tracking system.

4. The method for constructing a bipolar disorder experimental animal model according to claim 2, characterized in that: The method for the elevated cross maze experiment is as follows: The elevated cross maze experimental device consists of a white plexiglass box with two open arms and two closed arms, arranged in a cross shape, 40-60 cm above the ground; the experimental animal is placed alone in the center of the elevated cross maze experimental device, facing one of the closed arms, and is allowed to freely explore the device for 5 minutes; during the test, a video tracking system is used to quantify the total distance the experimental animal moves, the number of times it enters the open arms, and the time it spends on the open arms.

5. The method for constructing a bipolar disorder experimental animal model according to claim 1, characterized in that: The experimental animals were rodents.

6. An apparatus for constructing a biphasic disorder experimental animal model according to any one of claims 1 to 5, characterized in that: The device includes a rotating platform and a housing. The housing is positioned above the rotating platform, which has a switch. A touch bar is radially positioned on the lower part of the inner wall of the housing, adjacent to the upper surface of the rotating platform, at a distance of 5-10 mm. Resting bars A and B are also radially staggered on the inner wall of the housing, with resting bar A located 3-5 cm above the rotating platform and resting bar B located 7-10 cm above the platform. The continuous rotation of the platform and the obstruction of the touch bars disrupt the sleep of the experimental animals. The resting bars above the rotating platform allow for brief rests without preventing the animals from entering REM sleep, which would otherwise cause them to fall due to muscle relaxation. The top of the housing has an openable mesh, and a water bottle is located on the side. The rotating platform is connected to an intelligent circulation controller, which has a display screen and buttons.

7. The use of the animal model obtained by the method of any one of claims 1 to 5 in evaluating drugs that can alleviate or treat bipolar disorder.

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