Construction method, device and application of bipolar disorder experimental animal model

Abstract

The invention relates to the field of medical biology, and discloses a construction method and device of a bipolar disorder experimental animal model and application, the construction method comprises the following steps: selecting a healthy experimental animal to carry out environmental adaptation feeding before modeling, then carrying out sleep deprivation stimulation for 3-5 days continuously, and carrying out sleep deprivation stimulation for 15-18 hours continuously and 6-9 hours for rest every day so as to freely eat and feed water; and after the sleep deprivation stimulation is finished, evaluating the surface validity of the disease model for the experimental animal by adopting behavioral test. The construction method provided by the invention is simple to operate and easy to repeat, can better simulate the disease of a clinical bipolar disorder patient, and provides a new model normal form for pathogenesis research and clinical drug screening of the bipolar disorder, and the disease model can be maintained for a long time.

Description

Technical Field

[0001] The present invention relates to the field of medical biology, and in particular to a method for constructing an animal model of mental illness, and in particular to a method, device and application of a bipolar disorder experimental animal model. Background Art

[0002] Bipolar disorder (BD), also known as manic-depressive illness, is a neuropsychiatric condition characterized by recurrent, intermittent, or cyclical episodes of mania (or hypomania) and depression. It often begins in adolescence or early adulthood and can severely impact patients' personal and social functioning. BD is characterized by both manic or hypomanic episodes (characterized by elevated mood, racing thoughts, and heightened volitional activity) and depressive episodes (characterized by depressed mood, slowed thinking, and decreased volitional activity). Its recurring and chronic nature makes its treatment challenging. Current research on BD primarily focuses on the depressive phase, while research on the manic phase is limited. One major reason for this is the lack of effective animal models for BD. Developing novel animal models for BD is crucial for understanding the pathogenesis of mania.

[0003] Rats and mice are commonly used as animal models for bipolar disorder, with mice being the most commonly used. The book "Rodent Models and Behavioral Experiments in Mental Disorders," edited by Cui Donghong, Xu Lin, et al. (2024), provides a detailed overview of various animal models for BD, primarily genetically engineered models (Clock gene mutation models, GSK3 overexpression models), drug-induced models (amphetamine-induced models, ouabain-induced models), and environmental stress models (sleep deprivation models). Genetically engineered models significantly impact the normal reproduction and growth of mice, and their breeding cycle is long, resulting in high time and economic costs. The modeling drug amphetamine is a controlled psychostimulant, and its purchase and use are subject to strict approval procedures and hazardous chemical management regulations, resulting in few domestic literature reports. The modeling drug ouabain, on the other hand, requires targeted administration, making the modeling procedure cumbersome. Deviations from the stereotactic injection site can easily lead to model instability. Furthermore, anesthesia and surgery can easily cause mouse mortality, increasing experimental costs. Reduced sleep need is one of the typical clinical manifestations of BD manic episodes. Sleep deprivation is also an environmental stressor for BD manic episodes, which can induce BD patients to switch from a depressive state to a manic state. The multi-platform water environment stress model is the most commonly used sleep deprivation model, but the behavioral changes in mice may not only be caused by sleep deprivation, but may also be affected by the combined influence of multiple factors such as water environment stress and restricted activity space. It is difficult to accurately attribute them to sleep deprivation itself. Animals stay in a multi-platform water environment for a long time, their bodies are in a state of stress, their immunity is reduced, and they are prone to various diseases. It may even cause mice to become infected with diseases or drown. This will not only increase the cost of the experiment, 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 (grant announcement date: November 14, 2017) discloses a device for establishing a sleep deprivation model in mice. The device comprises a box-shaped body filled with an appropriate amount of liquid that is chemically harmless to mice. The depth of the liquid does not exceed the body length of the mouse. Above the liquid surface, at least one support rod is positioned for the mouse to stand. A top cover is provided with a solid food storage area and a water supply area for the mouse to eat and drink normally. The top cover is located over the upper opening of the box-shaped body. This device exploits the fact that mice or rats are afraid of water and cannot fall asleep by standing on the horizontal support rods near the water surface for a long time. This device is suitable for establishing a sleep deprivation model for long-term experiments with strict time control. 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 turbid, degrading the water quality, and producing an unpleasant odor. Drinking the water can increase the risk of infection and discomfort in mice. Water environment stress will also increase confounding factors other than sleep deprivation stimulation. The above unfavorable factors will not only increase the experimental cost, but also affect the reliability of the experimental results, and are not suitable for long-term stable modeling.

[0005] Chinese utility model patent CN222193580U discloses a sleep deprivation device for small animals used in bioengineering. The device comprises a base, the upper end of which is provided with a bottom plate, the center of which is provided with a rotating shaft, the lower end of which is connected to a drive motor within the base; a disturbance rod is provided on the rotating shaft; the base is provided with a cylinder, the top of which is provided with a top cover, and the top cover is provided with air vents; a lighting lamp is provided on the bottom surface of the top cover or the side wall of the cylinder; a waterer and a feeder are provided on the outer wall of the cylinder; and the front of the base is provided with a touch screen. This device uses a disturbance rod instead of a water environment, which can avoid the adverse effects of the water environment on the animals. It is simple to operate, has few interference factors, and is suitable for long-term stable modeling. However, this device causes the animals to be constantly touched by the disturbance rod, thus maintaining a disturbed sleep state and preventing them from resting. Therefore, this disturbance device is often used to construct animal models of depression and is not suitable for constructing mouse models of bipolar disorder.

[0006] Furthermore, there is currently a lack of a unified paradigm for constructing bipolar disorder mouse models using sleep deprivation in a multi-platform aquatic environment. The book "Rodent Models and Behavioral Experiments in Mental Disorders," edited by Cui Donghong, Xu Lin, et al. (2024), provides a detailed overview of bipolar disorder model construction methods. The sleep deprivation modeling duration is 72 hours, while some published literature uses modeling durations ranging from 36 to 72 hours. These modeling paradigms are all acute, and once the BD model is successfully constructed, the model is maintained for only about 30 minutes, making it impossible to observe the behavioral and physiological indicators of the model animals for more than an hour, and thus unable to meet the needs of longer-term physiological testing. Summary of the Invention

[0007] In order to solve the problems in the prior art of bipolar disorder mouse models constructed using existing paradigms, such as short maintenance time, multiple confounding factors, high experimental costs, and poor accuracy of experimental results, the present invention provides a method, device, and application for constructing a bipolar disorder experimental animal model.

[0008] In order to achieve the above technical objectives, the technical solution adopted by the present invention is:

[0009] In the first aspect, the present application provides a method for constructing an experimental animal model of bipolar disorder, wherein healthy experimental animals are selected for environmental adaptation before modeling, and then subjected to sleep deprivation stimulation for 3 to 5 days, during 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, behavioral tests are performed on the experimental animals 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 in the morning of the next day.

[0011] As a preferred technical solution, sleep deprivation stimulation is carried out in a sleep deprivation device, which includes a rotating platform and a box. The box cover is arranged above the rotating platform. A touch rod and a rest rod are radially arranged between the inner walls of the box. The touch rod is adjacent to the upper surface of the rotating platform, and the rest rod is arranged above the touch rod at intervals. The sleep of the experimental animal is disturbed by the continuous rotation of the rotating platform and the obstruction of the touch rod.

[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 open field test and elevated plus maze test.

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

[0015] As a preferred technical solution, the method of the elevated plus maze experiment is as follows: the elevated plus maze experimental device consists of a white plexiglass box with two open arms and two closed arms, the two closed arms and the two open arms are arranged in a cross shape, 40 to 60 cm above the ground; the experimental animal is placed individually in the center of the elevated plus 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 moved by the experimental animal, the number of times it enters the open arm, and the time it stays in the open arm.

[0016] As a preferred technical solution, the experimental animals are rodents.

[0017] In the second aspect, the present application provides a sleep deprivation device, including a rotating platform and a box body, the box body cover is arranged above the rotating platform, a switch is provided on the rotating platform, a touch rod and a rest rod are radially provided between the inner walls of the box body, the touch rod is adjacent to the upper surface of the rotating platform, and the rest rod is arranged above the touch rod at intervals. The continuous rotation of the rotating platform and the obstruction of the touch rod interfere with the sleep of experimental animals; a hole net that can be opened and closed is provided on the top of the box body, a water bottle is provided on the side of the box body, the rotating platform is connected to an intelligent circulation controller, and the intelligent controller is provided with a display screen and buttons.

[0018] In a second aspect, the present application provides an application, namely, the application of the animal model obtained by the above-mentioned method of the present application in evaluating drugs that can alleviate or treat bipolar disorder.

[0019] The beneficial effects of the present invention are:

[0020] The method for constructing an experimental animal model of bipolar disorder proposed in the present 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 clinical drug screening.

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

[0022] The sleep deprivation device designed by the present invention utilizes a rotating platform instead of a water environment and a disturbance rod, and disturbs the sleep of animals through the rotating platform and the touch rod. At the same time, a rest rod, food and drinking water are provided above the rotating platform. This device can satisfy the experimental animals' need for a short rest while preventing them from entering rapid eye movement sleep, which would otherwise cause them to fall due to muscle relaxation. This device can also greatly reduce the death 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 a short modeling time. It can better simulate the slow progression of bipolar disorder and is particularly suitable for large-scale, high-throughput preclinical animal testing of new drugs, accelerating the drug development process.

[0023] The device designed by the present invention has a simple structure and low manufacturing cost, is suitable for large-scale use, and greatly improves scientific research efficiency and saves research costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without creative work.

[0025] Figure 1 It is a structural schematic diagram of the sleep deprivation device in the present invention.

[0026] Figure 2 It is an analysis of the weight changes, open field test and elevated plus maze test of model mice after sleep deprivation stimulation.

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

[0028] Figure 4 This is an analysis of the open field test and elevated plus maze test of mice in the control group, treatment group, model group and treatment / model group after the end of sleep deprivation stimulation.

[0029] Figure numerals: 1-rotating platform, 2-switch, 3-touch rod, 4-rest rod A, 5-rest rod B, 6-box, 7-hole net, 8-kettle, 9-controller, 10-display screen, 11-button, 12-jack. DETAILED DESCRIPTION

[0030] In order to make the key technical points, technical solutions and beneficial effects of the present invention more clearly understood, the present invention is described in detail with reference to the following drawings and embodiments. It should be understood that the embodiments are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0031] The experimental methods used in the following examples are conventional methods unless otherwise specified. Other materials and reagents used, unless otherwise specified, can be purchased from commercial sources. The animal experiments implemented have been approved by the Experimental Animal Ethics Committee of West China Hospital, Sichuan University (Ethics Registration Number: 20230504005).

[0032] A method for constructing an experimental animal model of bipolar disorder comprises the following steps:

[0033] (1) Experimental Animal Selection: Select a number of experimental animals that match the animal strain, sex, age, weight, etc. according to the experimental needs and randomly group them. The experimental animals are rodents, such as mice and rats.

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

[0035] (3) Sleep deprivation stimulation: At the beginning of modeling, the parameters of the sleep deprivation device are first set, including the device's power-on time, power-off time, number of cycles, etc. The parameters of one embodiment provided by the present invention are that the rotating platform of the sleep deprivation device runs continuously for 3 to 5 days, 15 to 18 hours a day, the running time section is from 16:00 in the afternoon to 10:00 the next morning, and rests for 6 to 9 hours, and the rotating platform speed is 2 to 30 seconds / circle; then, the experimental animals are placed in the box of the sleep deprivation device in advance, covered with a mesh, and food and drinking water are added; the experimenter observes the state of the experimental animals, replenishes food, and replenishes water every other day; preferably, the experimental animals are sleep deprived for 16 hours and rest for 8 hours a day, and the modeling duration is 4 days.

[0036] (4) Behavioral evaluation of experimental animals in bipolar disorder models: Behavioral tests were used to evaluate the face validity of the disease model, including open field test and elevated plus maze test.

[0037] (5) Verification of the experimental animal model of bipolar disorder: Valproate (the first-line clinical drug for bipolar disorder) was used to evaluate the predictive validity of the disease model. The dose of valproate was 200 mg / kg, and the administration method was intraperitoneal injection once a day for a period of 7 days. After the completion of valproate treatment, 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: it includes a rotating platform 1, which is provided with a switch 2, which is used to control the power on of the device and the rotation direction of the rotating platform; it also includes a cylindrical box 6, which is covered above the rotating platform 1, and a touch rod 3 is radially provided on the lower part of the inner wall of the box 6. The touch rod 3 is adjacent to the upper surface of the rotating platform 1, and the touch rod 3 is 5 to 10 mm away from the upper surface of the rotating platform 1. The inner wall of the box 6 is also radially staggered with a rest rod A4 and a rest rod B5. The rest rod A4 is located 3 to 5 cm above the rotating platform 1, and the rest rod B5 is located 7 to 10 cm above the rotating platform 1. A hole mesh 7 that can be opened and closed is provided on the top of the box 6, and the hole mesh 7 is spaced 1 cm apart. The hole mesh 7 has both ventilation and feeding trough functions. A drinking bottle 8 is provided on the side of the box 6. The rotating platform 1 is connected to an intelligent circulation controller 9, which consists of a display screen 10, a button 11 and a jack 12. Preferably, the rotating platform 1 has a diameter of 14 cm, the box 6 is made of transparent material, the inner diameter of the box 6 is 15 cm, and the height is 15 cm; the size of the box and the rotating platform can also be customized according to the body size of mice and rats, so as to be used for sleep deprivation stimulation modeling of experimental animals of different body sizes.

[0039] Example 1

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

[0041] (1) Mouse selection: SPF-grade, untreated mice 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 g. The purchased mice were kept in a clean environment with a light cycle of 7:00-19:00.

[0042] (2) Pre-modeling adaptation: The mice were first fed and conditioned for 7 days. The mice were allowed to eat and drink freely. The experimenters observed the mice's condition, supplemented their food, and replenished their water every other day. The mice's weight was recorded before modeling. The mice were randomly divided into two groups: a control group (9 mice) and a model group (9 mice).

[0043] (3) Sleep deprivation stimulation: At the beginning of modeling, first set up the sleep deprivation device (such as Figure 1) parameters, the device was turned on from 18:00 every evening to 10:00 the next morning (continuous operation for 16 hours), that is, the rotating platform of the sleep deprivation device continued to operate for 16 hours, and the rotating platform speed was 30 seconds / circle; then the mice were placed in the sleep deprivation device box in advance (before 18:00), covered with a perforated mesh, and food and drinking water were added; the experimenters observed the status of the mice, supplemented food, and replenished water every other day; the mice were sleep deprived for 16 hours and rested for 8 hours every day, and the modeling lasted for 4 days.

[0044] (4) Behavioral evaluation of bipolar disorder model mice: After modeling, an open field test was conducted. Specifically, the open field test device was a box made of white plexiglass (40 cm × 40 cm × 35 cm). Each mouse was placed in the corner of the field and allowed to freely explore the device for 5 minutes. After each test, the open field test device was cleaned with 75% alcohol solution to remove any traces of odor. During this 5-minute test, the total movement distance, frequency of entering the central area, and time spent in the central area of ​​the mouse were analyzed by a video tracking system (Noldus, the Netherlands). Then, an elevated plus maze test was conducted. Specifically, the elevated plus maze test device 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 the two open arms were arranged in a cross shape and 60 cm above the ground. A camera was fixed above the elevated plus maze to record the activities of the mice. Each mouse was individually placed in the center of the elevated plus maze, facing one of the closed arms, and then allowed to freely explore the apparatus for 5 minutes. After each trial, the apparatus was cleaned with a 75% alcohol solution to remove any traces of odor. During the 5-minute test, a video tracking system was used to quantify the total distance traveled by the mouse, the number of arm entries, and the time spent in the open arms.

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

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

[0047] The experimental results are as follows Figure 2 As shown in the results, sleep deprivation stimulation can significantly reduce the weight of mice (P<0.01), and significantly increase the movement distance, central area residence time and frequency of mice in the open field (P<0.01), as well as increase the movement distance, open arm residence time and frequency in the elevated plus 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 Example 2 provides an analysis of the maintenance time of a bipolar disorder mouse model.

[0050] (1) Mouse selection: SPF-grade, untreated mice 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 g. The purchased mice were kept in a clean environment with a light cycle of 7:00-19:00.

[0051] (2) Pre-modeling adaptation: The mice were first fed and conditioned for 7 days. The mice were allowed to eat and drink freely. The experimenters observed the mice's condition, supplemented their food, and replenished their water every other day. The mice's weight was recorded before modeling. 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 modeling, first set up the sleep deprivation device (such as Figure 1 ) parameters, the device was turned on from 18:00 every evening to 10:00 the next morning (continuous operation for 16 hours), that is, the rotating platform of the sleep deprivation device continued to operate for 16 hours, and the rotating platform speed was 30 seconds per circle; then the mice were placed in the sleep deprivation device box in advance (before 18:00), covered with a mesh, and food and drinking water were added; the experimenters observed the status of the mice, supplemented food, and replenished water every other day; the mice were sleep deprived for 16 hours and rested for 8 hours every day, and the modeling lasted for 4 days.

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

[0054] Control group: Mice were placed in cages without any treatment and directly underwent open field test.

[0055] Model group - 1h: Mice were treated according to the above step (3), and an open field test was performed 1 hour after the model was established.

[0056] Model group - 4h: Mice were treated according to the above step (3), and an open field test was performed 4 hours after the model was established.

[0057] Model group - 12h: Mice were treated according to the above step (3), and an open field test was performed 12 hours after the model was established.

[0058] Model group - 24h: Mice were treated according to the above step (3), and an open field test was performed 24 hours after the model was established.

[0059] The experimental results are as follows Figure 3 As shown, the results showed that sleep deprivation stimulation could significantly increase the activity of mice (P<0.01), showing manic-like behavior. The model could be maintained for more than 4 hours, which could meet the requirements for long-term observation of the behavioral and physiological indicators of model animals.

[0060] Example 3

[0061] This Example 3 provides a method for constructing a bipolar disorder mouse model and evaluating the predictive validity of the model.

[0062] (1) Mouse selection: SPF-grade, untreated mice 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 g. The purchased mice were kept in a clean environment with a light cycle of 7:00-19:00.

[0063] (2) Pre-modeling adaptation: The mice were first fed and acclimated to the modeling environment for 7 days. The mice were allowed to eat and drink freely. The experimenters observed the mice's condition, supplemented food and water every other day, and randomly divided the mice 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 modeling, first set up the sleep deprivation device (such as Figure 1 ) parameters, the device was turned on from 18:00 every evening to 10:00 the next morning (continuous operation for 16 hours), that is, the rotating platform of the sleep deprivation device continued to operate for 16 hours, and the rotating platform speed was 30 seconds / circle; then the mice were placed in the sleep deprivation device box in advance (before 18:00), covered with a mesh, and food and drinking water were added; the experimenters observed the status of the mice, supplemented food, and replenished water every other day; the mice were sleep deprived for 16 hours and rested for 8 hours every day, and the modeling lasted for 4 days.

[0065] (4) Behavioral evaluation of bipolar disorder model mice: After modeling, an open field test was conducted. Specifically, the open field test device was a box made of white plexiglass (40 cm × 40 cm × 35 cm). Each mouse was placed in the corner of the field and allowed to freely explore the device for 5 minutes. After each test, the open field test device was cleaned with 75% alcohol solution to remove any traces of odor. During this 5-minute test, the total movement distance, frequency of entering the central area, and time spent in the central area of ​​the mouse were analyzed by a video tracking system (Noldus, the Netherlands). Then, an elevated plus maze test was conducted. Specifically, the elevated plus maze test device 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 the two open arms were opposite each other and arranged in a cross shape, 60 cm above the ground. A camera was fixed above the elevated plus maze to record the activities of the mice. Each mouse was individually placed in the center of the elevated plus maze, facing one of the closed arms, and then allowed to freely explore the apparatus for 5 minutes. After each trial, the apparatus was cleaned with a 75% alcohol solution to remove any traces of odor. During the 5-minute test, a video tracking system was used to quantify the total distance traveled by the mouse, the number of arm entries, and the time spent in the open arms.

[0066] Control group: Mice were placed in a cage and treated in parallel with the control group, and then subjected to open field test and elevated plus maze test.

[0067] Treatment group: Mice were placed in cages, and valproate (a first-line clinical drug for bipolar disorder) was used to evaluate the predictive validity of the disease model. The valproate dosage was 200 mg / kg, and the administration method was intraperitoneal injection, once a day, for a dosing cycle of 7 days. After the completion of valproate treatment, open field test and elevated plus maze test were performed.

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

[0069] Treatment / model group: Mice were treated according to the above step (3) and valproate treatment at the same time. After treatment, open field test and elevated plus maze test were performed.

[0070] The experimental results are as follows Figure 4As shown, the results showed that sleep deprivation stimulation can significantly increase the movement distance, central area residence time and frequency of mice in the open field (P<0.01), and increase the movement distance, open arm residence time and frequency in the elevated plus maze (P<0.05), while valproate (the first-line clinical drug for bipolar disorder) treatment can significantly reverse the above changes in model mice (P<0.001). The experimental results have confirmed that the new bipolar disorder mouse model proposed in the present invention has passed the predictive validity evaluation.

[0071] The above results indicate that the method for constructing a bipolar disorder mouse model provided by the embodiments of the present invention can quickly obtain an ideal mouse model.

[0072] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent replacement or improvement made within the principles and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for constructing an experimental animal model of bipolar disorder, characterized by: Healthy experimental animals were selected for environmental adaptation before modeling, and then 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. After the sleep deprivation stimulation, behavioral tests were performed on the experimental animals to evaluate the face validity of the disease model.

2. The method for constructing an experimental animal model of bipolar disorder according to claim 1, characterized in that: The sleep deprivation period is 15 to 18 consecutive hours from 16:00 in the afternoon to 10:00 in the morning of the next day.

3. The method for constructing an experimental animal model of bipolar disorder according to claim 2, characterized in that: Sleep deprivation stimulation is carried out in a sleep deprivation device, which includes a rotating platform and a box. The box cover is located above the rotating platform. Touch rods and rest rods are radially arranged between the inner walls of the box. The touch rods are adjacent to the upper surface of the rotating platform, and the rest rods are spaced above the touch rods. The sleep of experimental animals is disturbed by the continuous rotation of the rotating platform and the obstruction of the touch rods.

4. The method for constructing an experimental animal model of bipolar disorder according to claim 3, characterized in that: The rotation speed of the rotating platform is 2 to 30 seconds per revolution.

5. The method for constructing an experimental animal model of bipolar disorder according to claim 1, characterized in that: Behavioral tests included the open field test and the elevated plus maze test.

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

7. The method for constructing an experimental animal model of bipolar disorder according to claim 5, characterized in that: The method of the elevated plus maze experiment is as follows: the elevated plus maze experimental apparatus consists of a white plexiglass box with two open arms and two closed arms. The two closed arms and two open arms are arranged in a cross shape and are 40 to 60 cm above the ground; the experimental animal is placed individually in the center of the elevated plus maze apparatus, facing one of the closed arms, and allowed to freely explore the apparatus for 5 minutes; during the test, a video tracking system is used to quantify the total distance moved by the experimental animal, the number of times it enters the open arm, and the time it stays in the open arm.

8. The method for constructing an experimental animal model of bipolar disorder according to claim 1, characterized in that: The experimental animals are rodents.

9. A device for the method for constructing a bipolar disorder experimental animal model according to any one of claims 1 to 8, characterized in that: It includes a rotating platform and a box body. The box body cover is arranged above the rotating platform. A switch is provided on the rotating platform. A touch rod and a rest rod are radially provided between the inner walls of the box body. The touch rod is adjacent to the upper surface of the rotating platform. The rest rod is arranged above the touch rod at intervals. The continuous rotation of the rotating platform and the obstruction of the touch rod interfere with the sleep of the experimental animals. A hole net that can be opened and closed is provided on the top of the box body. A water bottle is provided on the side of the box body. The rotating platform is connected to an intelligent circulation controller. The intelligent controller is provided with a display screen and buttons.

10. Use of the animal model obtained by the method according to any one of claims 1 to 8 in evaluating drugs that can alleviate or treat bipolar disorder.

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