A method for intervening in the behavior of autistic mice based on terahertz waves and application thereof

Abstract

The application provides a method for treating autism spectrum disorder (ASD) based on terahertz wave intervention and application thereof. The method comprises the following steps: dividing mice into four groups according to groups, each group having two types of mice; testing baseline behaviors of anxiety, motor ability and social ability of each group of mice; after a set time of completion of baseline behavior testing, using a THz-VNS device to intervene in each group of mice by THz-VNS; after the mice wake up, behavior testing is performed after a set time of completion of THz-VNS intervention; and statistically analyzing the behavior testing results to obtain the intervention results of THz-VNS intervention on ASD. By comparing the behavior testing results of wild type and ASD model mice before and after intervention, it is proved that the intervention method can improve the anxiety abnormality, motor dysfunction and social dysfunction of ASD model mice, which indicates that the intervention method can be used for the treatment of ASD. Meanwhile, the intervention method has a wide influence on the nervous system function, so the intervention is not limited to the treatment of ASD, but can also be extended to various nervous system diseases.

Description

Technical Field

[0001] This invention belongs to the field of neurobiological experimental technology, specifically relating to a method and application of terahertz wave-based intervention on the behavior of autistic mice. Background Technology

[0002] Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by impairments in social interaction, restricted interests, and repetitive, stereotyped behaviors. In recent years, its global prevalence has been steadily increasing, seriously harming children's physical and mental health and imposing a heavy burden on families and society. In addition to the typical core symptoms, affected children often exhibit accompanying symptoms such as language delay, motor disorders, epilepsy, hyperacusis, sensory abnormalities, sleep disturbances, mood swings, feeding difficulties, and indigestion, which undoubtedly exacerbates the complexity and challenge of clinical intervention for ASD. Currently, while ASD cannot be completely cured, standardized medical intervention and systematic rehabilitation training can significantly improve patients' quality of life and social functioning. Besides traditional drug treatments (such as risperidone and aripiprazole) and behavioral training interventions, new intervention strategies such as targeted drug therapy, stem cell and exosome therapy, gene therapy, transcranial magnetic stimulation, and virtual reality (VR) / robot-assisted therapy have also shown potential clinical application value in recent years. However, these intervention strategies generally have limitations such as high treatment costs, limited application scope, and high risk of side effects. Therefore, most new intervention strategies are still in the experimental or clinical validation stage and are difficult to popularize.

[0003] Vagus nerve stimulation (VNS) is a neuromodulation technique that modulates brain function by electrically stimulating the vagus nerve. Currently, there are two main forms: implantable VNS, which involves surgically implanting electrodes and a pulse generator to stimulate the cervical vagus nerve, and percutaneous VNS (tVNS), which stimulates the auricular branch of the vagus nerve through the auricle. Clinical studies have shown that implantable VNS can improve social, language, and emotional behaviors in children with epilepsy and ASD, independent of epilepsy control, suggesting that VNS itself may have a positive effect on the core symptoms of ASD. On the other hand, animal studies have shown that repeated tVNS can significantly improve social and cognitive functions and reduce social anxiety and general anxiety-like behaviors in ASD model mice. These results suggest that VNS treatment for ASD has certain application value. However, implantable VNS devices are expensive, carry the risk of surgical complications, and are mainly suitable for ASD patients with refractory epilepsy. tVNS stimulation intensity is limited, penetration depth is limited, repeated stimulation is required, it can cause ear discomfort, and its clinical efficacy is mild and varies greatly from person to person. Therefore, VNS cannot yet replace traditional behavioral interventions and drug treatments.

[0004] Terahertz (THz) waves are electromagnetic waves with frequencies ranging from 0.1 to 10 THz (wavelengths approximately 3 mm to 30 μm), falling between microwaves and infrared light. In recent years, terahertz intervention technology, as an emerging neuromodulation technique, has demonstrated unique advantages in the biomedical field, particularly in neuromodulation and the treatment of brain diseases. Studies have shown that terahertz waves can non-invasively, non-thermally, and reversibly modulate neuronal activity, while also promoting nerve growth, enhancing synaptic transmission, improving mitochondrial function, and exhibiting anti-inflammatory and antioxidant effects, suggesting potential applications in the treatment of acute myeloid sclerosis (ASD). Although the mechanism demonstrates potential feasibility, stimulation parameters and safety require further verification, and there are currently no studies on its use in the treatment or intervention of ASD.

[0005] Combining the advantages and disadvantages of the two intervention methods mentioned above, using THz wave stimulation to replace the electrical stimulation in tVNS technology creates a terahertz-vagus nerve stimulation (THz-VNS) intervention, which theoretically may produce a better synergistic intervention effect. However, there are currently no relevant clinical studies on THz wave stimulation of the vagus nerve for the treatment of ASD, and the safety and effectiveness of this combined intervention method still need to be verified through rigorous animal experiments and phased clinical trials. Summary of the Invention

[0006] To address the problems existing in the prior art, this invention provides a method and application for intervening in the behavior of autistic mice based on terahertz waves. It constructs an intervention method for terahertz wave stimulation of the vagus nerve (THz-VNS) and tests it on representative ASD model animals. Shank3b We investigated various behaviors in knockout mice before and after intervention, evaluated the therapeutic effect of combined intervention on ASD, and explored the underlying neural mechanisms.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is: a method for intervening in the behavior of autistic mice based on terahertz waves, comprising the following steps: The mice were divided into four groups, with two types of mice in each group. Baseline behaviors of anxiety, motor ability, and social ability were tested in each group of mice; After the baseline behavioral test was completed within the set time, each group of mice underwent THz-VNS intervention using a THz-VNS device. After the THz-VNS intervention was completed and the mice were awakened, behavioral tests were conducted at set times. Statistical analysis of the behavioral test results yielded the intervention results of THz-VNS intervention on ASD.

[0008] Furthermore, the four groups are: wild-type and blank stimulation group, wild-type and THz stimulation group, Shank3b Knockout and blank stimulus group Shank3b Knockout and THz-VNS stimulation group.

[0009] Furthermore, the THz-VNS device includes a terahertz wave source, a beam transmission and control module, and a small animal anesthesia system. The terahertz wave source is a quantum cascade laser with a center frequency of 50.47-56.04 THz, a wavelength of 5.35-5.94 µm, a pulse width of 171 ns, a maximum average output power of approximately 176 mW, and a peak power of approximately 1.03 W. The beam transmission and control module includes a polycrystalline infrared fiber, a polarization filter, a phase matching device, and a precision translation stage. The small animal anesthesia system is an inhalation-type gas anesthesia machine.

[0010] Further, THz-VNS intervention includes: Anesthetize the mice; The control system of the terahertz wave source will be turned on, the center wavelength and center frequency of the terahertz wave will be set, the power of the output end of the polycrystalline infrared fiber will be detected using a mid-infrared terahertz wave detector, and the output power will be adjusted to the set power by fine-tuning the precision translation stage; then, the output end of the polycrystalline infrared fiber will be fixed to the concha of the mouse auricle using an ear clip, and the control system will output an on signal to perform THz-VNS intervention according to the set parameters.

[0011] Furthermore, anesthetizing the mice includes: turning on the small animal inhalation gas anesthesia machine, setting the anesthetic output concentration and oxygen flow rate, placing the mice in the anesthesia induction box, and after the mice enter an unconscious state, removing them and placing them on a constant temperature mat and putting on an inhalation anesthesia mask. Furthermore, the specific THz-VNS intervention parameters are: duration of 15 min, pulse width of 17 ns, repetition frequency of 1 MHz, and duty cycle of 40%.

[0012] Furthermore, the anxiety test in mice was conducted in an open field experiment for 5 minutes; the motor ability test in mice was conducted in an open field experiment for 30 minutes; and the social ability test in mice was conducted in a three-box social experiment for 10 minutes.

[0013] Furthermore, the behavioral testing methods were the same before and after the THz-VNS intervention.

[0014] Furthermore, THz-VNS intervention and behavioral training are carried out simultaneously, with behavioral training conducted before THz-VNS intervention, to achieve multimodal physical intervention combined with behavioral training.

[0015] The present invention also provides an application method for the terahertz wave-based vagus nerve stimulation intervention to improve the behavior of autistic mice as described above, for evaluating the intervention results of THz-VNS intervention on anxiety, motor ability and social ability in ASD mice, and based on the intervention results, providing data for elucidating the underlying central and peripheral neural circuit mechanisms, for the intervention and treatment of other accompanying symptoms of ASD.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: Taking advantage of the superficial nature of the vagus nerve branches under the auricle skin, terahertz waves are used to percutaneously stimulate the vagus nerve. Through the comprehensive biological effects of terahertz waves, including promoting nerve growth, enhancing synaptic transmission, and promoting the resonance of biomolecules, a complete micro-macro-regulatory chain is formed. THz waves regulate neuronal excitability and energy metabolism at the cellular and molecular level, thereby regulating brain network function at the nervous system level. Secondly, THz waves can promote nerve growth, enhance synaptic transmission, and improve mitochondrial function. THz-VNS can enhance brain network flexibility, regulate neurotransmitter release, and promote neural plasticity. Thirdly, although THz-VNS is applied locally, it widely affects multiple brain regions through the vagus nerve-brain axis, producing immediate and cumulative synergistic effects. It is safe and tolerable, and because it is non-invasive, the risks are controllable.

[0017] Given the aforementioned advantages, THz-VNS can be used to treat and improve various behaviors associated with ASD. This intervention can activate vagal afferent fibers by modulating ion channels, enhancing mitochondrial function, and / or strengthening synaptic function, thereby affecting the function of brain regions such as the prefrontal cortex, anterior cingulate cortex, motor cortex, amygdala, and thalamus. Improvements in the function of these brain regions can, to some extent, salvage behaviors associated with ASD, such as social impairments, motor disorders, and anxiety.

[0018] THz-VNS may also improve other accompanying symptoms of ASD, such as language delay, epilepsy, sleep disorders, eating difficulties, and indigestion, many of which are highly correlated with vagus nerve function. Furthermore, this intervention may also help improve neurological disorders such as epilepsy, depression, Alzheimer's disease, and post-traumatic stress disorder. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the THz-VNS device based on the present invention.

[0020] Figure 2 Wild type and Shank3b Changes in social behavior, motor behavior, and anxiety in knockout mice before and after THz-VNS intervention. Detailed Implementation

[0021] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] This invention provides a terahertz wave-based vagus nerve stimulation intervention and its application in improving the behavior of autistic mice, including the following: The THz-VNS-based device comprises four parts: a terahertz wave source, a beam transmission and control module, a small animal anesthesia system, and a control system. The terahertz wave source is a quantum cascade laser (QCL) capable of generating a center frequency of 50.47-56.04 THz, a wavelength of 5.35-5.94 µm, a pulse width of 171 ns, a maximum average output power of approximately 176 mW, and a peak power of approximately 1.03 W. The beam transmission and control module includes a beam control component built into the wave source (for controlling the polarization and directivity of the wave), a focusing lens (for focusing or shaping the terahertz wave), a precision translation stage (for accurately guiding the focused or shaped terahertz wave generated by the QCL to the target position), and a polycrystalline infrared fiber (PIR) with a diameter of 0.5 ± 0.05 mm. The precision translation stage is a three-axis (X, Y, and Z) moving platform, and the control system can control the switching of the QCL and adjust its output parameters. In summary, the terahertz waves emitted by the quantum cascade laser are linearly polarized by the beam control component built into the wave source, and then precisely transmitted to one end of the connected polycrystalline infrared fiber via a focusing lens and a precision translation stage. The other end of the polycrystalline infrared fiber acts on a predetermined site on the mouse's body. The small animal anesthesia system is an inhalation-type gas anesthesia machine, which allows for rapid induction and precise control of the depth of anesthesia in small animals. This ensures that the animal enters a stable, unconscious state during the terahertz wave combined with vagus nerve stimulation intervention, avoiding intervention deviations and equipment damage caused by the animal struggling and escaping while conscious.

[0023] Intervention and behavioral testing procedures: Before the experiment, 8-week-old mice were divided into 4 groups of 4 mice each, namely wild-type + blank stimulus, wild-type + THz-VNS stimulus, etc. Shank3b Knockout + blank stimulus Shank3b Knockout + THz-VNS stimulation. Shank3b Knockout mice are a typical application, but other different autism mouse models or sensory disorder mouse models can be used depending on the experimental purpose.

[0024] Before THz-VNS intervention in mice, baseline behaviors were assessed using behavioral tests, including anxiety (open field test, 5 min), motor skills (open field free exploration, 30 min), and social skills (three-box socialization test, 10 min). The interval between the two behavioral tests was 2 hours. Three days after the baseline behavioral tests, THz-VNS intervention began. First, the small animal inhalation gas anesthesia machine was turned on, and the isoflurane output concentration was set (4% during induction and 1.5% during maintenance) and oxygen flow rate was set to 0.5 L / min. Mice were placed in the anesthesia induction box, and after entering an unconscious state, they were quickly removed, placed on a temperature-controlled mat, and fitted with an inhalation anesthesia mask. Second, the PIR and QCL were connected, the control system was turned on, and the terahertz center wavelength was set to approximately 5.4063 µm, with a center frequency of [missing information]. At 55.46 THz, the PIR output power was detected using a mid-infrared terahertz wave detector, and the output power was adjusted to 5 mW using a fine-tuning precision translation stage. Next, the PIR output was fixed to the concha region of the mouse auricle using a self-made ear clip, and the intervention was initiated by outputting an on signal through the control system. The intervention lasted for 15 min, with a pulse width of 171 ns, a repetition frequency of 1 MHz, and a duty cycle of 40%. Finally, 30 min after the mice recovered from the intervention, the post-intervention behavioral test was started, and the experimental procedure was the same as the baseline behavioral test.

[0025] The behavioral test results were statistically analyzed using EthoVision XT 15 analysis software. The results showed that mice in each group before and after terahertz wave-based vagus nerve stimulation intervention had different levels of anxiety (percentage of time spent in the central region), motor ability (total distance traveled), and social ability (social preference score). This was used to evaluate the therapeutic effect of THz-VNS intervention on ASD.

[0026] By comparing the behavioral test results of wild-type and ASD model mice before and after the intervention, it was demonstrated that this intervention method can improve anxiety, motor disorders, and social impairments in ASD model mice, suggesting that this intervention method can be used for the treatment of ASD. Figure 2 Furthermore, this intervention method has a wide-ranging impact on nervous system function, so it is not limited to the treatment of ASD and can be extended to various neurological diseases. On the other hand, this intervention method does not require invasive surgery, is simple to perform, has a short intervention period, a non-overlapping spectrum of adverse reactions, and a high safety profile.

[0027] Example 1, The anxiety behavior testing device consisted of a 30×30×40cm open-field enclosure and an automated tracking and recording system. Before the test, mice were transferred to the behavioral laboratory to acclimatize to the environment for 1 hour at a temperature of 24±1℃ and a light intensity of 10 lux. At the start of the test, each mouse was gently placed in the enclosure and allowed to explore freely for 5 minutes, while its movement was automatically tracked and recorded. The total distance traveled and the percentage of time spent in the central area were analyzed using an EthoVision XT 15 to assess the mice's anxiety levels; a decrease in the percentage of time spent in the central area indicated abnormal anxiety. (Reference) Figure 2 Choose A and B, and the results are as follows: (1) After THz-VNS intervention Shank3b The knockout mice spent a significantly increased percentage of time in the central region, while wild-type mice showed no significant change, suggesting that THz-VNS intervention significantly improved anxiety in ASD mice, but had no significant effect on wild-type mice. (2) After THz-VNS intervention Shank3b The total distance traveled by both knockout and wild-type mice was not significantly different, suggesting that THz-VNS intervention had no significant effect on the physiological state of wild-type and ASD mice.

[0028] Example 2 The behavioral testing device for motor ability consisted of a 30×30×40cm familiarization box and an automated tracking and recording system. Before the test, mice were transferred to the behavioral laboratory to acclimatize for 1 hour at a temperature of 24±1℃ and a light intensity of 10 lux. At the start of the test, each mouse was gently placed in the testing box and allowed to explore freely for 30 minutes, with its movement trajectory automatically tracked and recorded. The total distance and cumulative duration of movement for each mouse were analyzed using an EthoVision XT 15 to assess its motor ability; a decrease in total distance and cumulative duration indicated a degree of motor impairment. (Reference) Figure 2 Choose C and D, and the results are as follows: (1) After THz-VNS intervention Shank3b The total distance of movement in knockout mice increased significantly, while there was no significant change in wild-type mice, suggesting that terahertz wave combined with vagus nerve stimulation intervention has a significant effect on improving motor disorders in ASD mice, but has no significant effect on wild-type mice. (2) After THz-VNS intervention Shank3b Knockout mice showed a significant increase in cumulative movement time, while wild-type mice showed no significant change, further suggesting that THz-VNS intervention has a significant ameliorative effect on motor dysfunction in ASD mice, but no significant effect on wild-type mice.

[0029] Example 3 The social competence behavioral testing device consisted of a three-box setup measuring 40×20×24cm and an automatic tracking and recording system. Each of the left and right boxes contained a transparent fence for placing a test mouse, while the middle box had barriers that allowed / restricted animal entry into the side boxes. Before the test, the mice were transferred to the behavioral laboratory to acclimatize to the environment for 1 hour at a temperature of 24±1℃ and a light intensity of 10 lux. At the start of the test, the test mouse was placed in the transparent fence of one of the boxes. Afterward, the experimental animal was placed in the middle box for 3-5 minutes to acclimatize. Simultaneously, the barriers on both sides were opened, allowing the mouse to explore freely for 10 minutes, and its movement was automatically tracked and recorded. The total distance each mouse traveled and the cumulative time spent exploring both the empty fence box and the tool mouse fence box were analyzed using an EthoVision XT 15. A social preference score was calculated as follows: (Cumulative time spent exploring the tool mouse fence box - Cumulative time spent exploring the empty fence box) / (Cumulative time spent exploring the tool mouse fence box + Cumulative time spent exploring the empty fence box). This score was used to assess the mice's social abilities; a decreased social preference score indicated a degree of social impairment. (Reference) Figure 2 For E and F, the results are as follows: (1) After THz-VNS intervention Shank3b The social preference scores of knockout mice were significantly increased, while those of wild-type mice showed no significant change, suggesting that THz-VNS intervention has a significant effect on improving social impairment in ASD mice, but has no significant effect on wild-type mice. (2) After THz-VNS intervention Shank3b The total distance traveled by both knockout and wild-type mice was not significantly different, suggesting that THz-VNS intervention had no significant effect on the physiological state of wild-type and ASD mice.

[0030] Therefore, from Figure 2 From A, we can see that the results of mice staying in the central region of an open field show that after THz-VNS intervention... Shank3b The knockout group mice showed a significantly increased percentage of time spent in the central region, while the other three groups showed no significant change before and after intervention; from Figure 2 As can be seen from B, there was no significant difference in the total distance moved by mice in the open field before and after the intervention in each group; Figure 2 The middle C represents the cumulative exercise duration in mice during the exercise experiment, showing the effect of THz-VNS intervention. Shank3b The knockout group mice showed a significant increase in cumulative exercise time, while the other three groups showed no significant changes before and after intervention; from Figure 2 The results of the THz-VNS intervention show that the total distance moved by mice in the exercise experiment was obtained from the D-D method. Shank3bThe total distance traveled by the knockout group mice was significantly increased, while there were no significant changes before and after intervention in the other three groups; E data showed that in the three-box social experiment, the social preference results of the mice were significantly improved after THz-VNS intervention. Shank3b Mice in the knockout group showed a significant increase in social preference scores, while the other three groups showed no significant changes before and after intervention; in group F, there were no significant differences in the total distance moved by mice in the three social apparatuses before and after intervention. Statistical graphs are presented as mean ± standard error, *: p <0.05, **: p <0.01, ns: no statistically significant difference.

[0031] In summary, this invention provides a method for intervening in the behavior of autistic mice based on terahertz waves. Mice are divided into four groups, with two types of mice in each group. Baseline behaviors of anxiety, motor skills, and social skills are tested in each group. After a set time following the completion of the baseline behavior test, each group of mice undergoes THz-VNS intervention using a THz-VNS device. After the mice recover from the THz-VNS intervention, behavioral tests are conducted again at a set time. Statistical analysis of the behavioral test results yields the intervention results of THz-VNS on ASD. Based on the method and results of THz-VNS intervention in improving the behavior of autistic mice described in this invention, researchers can evaluate the therapeutic effects of this intervention on anxiety, motor skills, and social skills in ASD mice and further elucidate the underlying central and peripheral neural circuit mechanisms, filling a gap in the field of neural modulation in ASD research and providing data support for future clinical applications. Furthermore, this method can be performed simultaneously with behavioral training, achieving multimodal physical intervention combined with behavioral training, which theoretically may produce better therapeutic effects. Considering the similar neuropathological mechanisms, this method can be further applied to the intervention and treatment of other accompanying symptoms of ASD. Furthermore, this method can be extended to the treatment of neurological diseases beyond ASD.

[0032] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A method for intervening in the behavior of autistic mice based on terahertz waves, characterized in that, Includes the following steps: The mice were divided into four groups, with two types of mice in each group. Baseline behaviors of anxiety, motor ability, and social ability were tested in each group of mice; After the baseline behavioral test was completed within the set time, each group of mice underwent THz-VNS intervention using a THz-VNS device. After the THz-VNS intervention was completed and the mice were awakened, behavioral tests were conducted at set times. Statistical analysis of the behavioral test results yielded the intervention results of THz-VNS intervention on ASD.

2. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, The four groups are: wild-type and blank stimulation group, wild-type and THz stimulation group, etc. Shank3b Knockout and blank stimulus group Shank3b Knockout and THz-VNS stimulation group.

3. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, The THz-VNS device includes a terahertz source, a beam transmission and control module, and a small animal anesthesia system. The terahertz source is a quantum cascade laser with a center frequency of 50.47-56.04 THz, a wavelength of 5.35-5.94 µm, a pulse width of 171 ns, a maximum average output power of approximately 176 mW, and a peak power of approximately 1.03 W. The beam transmission and control module includes a polycrystalline infrared fiber, a polarization filter, a phase matching device, and a precision translation stage. The small animal anesthesia system is an inhalation-type gas anesthesia machine.

4. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, THz-VNS intervention includes: Anesthetize the mice; The control system of the terahertz wave source will be turned on, the center wavelength and center frequency of the terahertz wave will be set, the power of the output end of the polycrystalline infrared fiber will be detected using a mid-infrared terahertz wave detector, and the output power will be adjusted to the set power by fine-tuning the precision translation stage; then, the output end of the polycrystalline infrared fiber will be fixed to the concha of the mouse auricle using an ear clip, and the control system will output an on signal to perform THz-VNS intervention according to the set parameters.

5. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 4, characterized in that, Anesthetizing mice involves: turning on the small animal inhalation gas anesthesia machine, setting the anesthetic output concentration and oxygen flow rate, placing the mouse in the anesthesia induction box, and after the mouse enters an unconscious state, removing it, placing it on a constant temperature mat, and putting an inhalation anesthesia mask on it.

6. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 4, characterized in that, The specific THz-VNS intervention parameters are: duration of 15 min, pulse width of 17 ns, repetition frequency of 1 MHz, and duty cycle of 40%.

7. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, The mouse anxiety test was conducted in an open field experiment with a set duration; the mouse motor ability test was conducted in an open field experiment with a set duration; and the mouse social ability test was conducted in a three-box social experiment with a set duration.

8. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, The behavioral testing methods were the same before and after the THz-VNS intervention.

9. The method for intervening in the behavior of autistic mice based on terahertz waves according to claim 1, characterized in that, THz-VNS intervention and behavioral training are carried out simultaneously. Behavioral training is conducted before THz-VNS intervention to achieve multimodal physical intervention combined with behavioral training.

10. The method of applying the terahertz wave-based vagus nerve stimulation intervention to improve the behavior of autistic mice as described in any one of claims 1-9, characterized in that, The purpose is to evaluate the effects of THz-VNS intervention on anxiety, motor function, and social function in ASD mice, and to provide data based on these results to elucidate the underlying central and peripheral neural circuit mechanisms for interventional treatment of other accompanying symptoms of ASD.

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