Method for analyzing influence of head-mounted focused ultrasound on mouse epileptic discharge

Abstract

The invention provides a mouse epilepsy research method based on head-mounted focused ultrasound equipment, which is used for evaluating the influence of focused ultrasound on neuron activity and epileptic seizure frequency in an epilepsy mouse model, and combines behavioristics, electrophysiology, calcium imaging and brain metabolism imaging technologies. The precise regulation and control effect of focused ultrasound on a target brain area is systematically studied, the head-mounted equipment comprises a micro transducer, a cooling system and an adjustable fixing device, flexible adjustment of frequency, power density and pulse parameters can be achieved, and experiments show that the frequency and duration of epileptic seizure are remarkably reduced through focused ultrasound stimulation, and the accuracy of the epileptic seizure is improved. The invention has important application value in the aspects of studying epilepsy pathology mechanism and developing novel non-invasive treatment technology, and provides theoretical basis and experimental support for clinical transformation of focused ultrasound.

Description

Technical Field

[0001] The present invention relates to a head-mounted focused ultrasound device and an application method thereof, and in particular to a method for analyzing the effects of focused ultrasound on neuronal activity and epileptic seizure frequency in an epileptic mouse model, and belongs to the fields of biomedical engineering and neuroscience. Background Art

[0002] Epilepsy is a chronic neurological disease that affects approximately 1% of the world's population. Its main feature is the abnormal discharge of brain neurons, which causes transient sensory, motor, consciousness or behavioral disorders. Currently, drug therapy is mainly used clinically to control epileptic seizures, but about 30% of patients are ineffective against drug therapy and are called refractory epilepsy. For these patients, surgery or neuromodulation technology becomes possible alternative therapies, but surgery is invasive and risky, and existing neuromodulation technologies (such as vagus nerve stimulation and deep brain stimulation) often require implanted devices and are costly.

[0003] As a non-invasive technology, focused ultrasound has shown great potential in the field of neuromodulation in recent years. It can regulate neural activity non-contactly through mechanical force or thermal effect, thereby achieving precise intervention in specific brain areas. Compared with traditional methods, focused ultrasound has the advantages of high controllability, non-invasiveness and precise positioning, and has achieved initial results in the research of Parkinson's disease, depression and other fields. However, there is a lack of systematic research on its mechanism of action and effect evaluation in the treatment of epilepsy.

[0004] Mouse epilepsy models are important tools for studying the pathogenesis and treatment of epilepsy, but due to the small size and complex brain structure of mice, existing focused ultrasound equipment and methods are difficult to achieve precise regulation of mouse brain regions. In addition, systematic methods for multimodal research in mouse models that combine behavioral analysis, electrophysiological recording, and imaging techniques are relatively limited.

[0005] Therefore, the development of a head-mounted focused ultrasound device suitable for mice and its application method are of great significance for exploring the mechanism of action of focused ultrasound in the treatment of epilepsy. Summary of the invention

[0006] The present invention provides a method for studying mouse epilepsy based on a head-mounted focused ultrasound device, which is used to evaluate the effects of focused ultrasound on epileptic seizure patterns, neuronal electrical activity and brain tissue metabolism. The present invention aims to provide an accurate, repeatable and easy-to-operate method to support the basic research and clinical application transformation of focused ultrasound in the treatment of epilepsy.

[0007] The technical solution of the present invention:

[0008] (1) Equipment design:

[0009] The head-mounted focused ultrasound device includes a miniature focused ultrasound transducer (frequency range is 0.5MHz to 3MHz, and focusing depth is adjustable), a positioning and fixing component (used to stably mount the transducer on the mouse's head), a control system (including a signal generator and a power regulation module), and a cooling system (to prevent local overheating of the mouse's head caused by long-term stimulation).

[0010] (2) Preparation of mouse model:

[0011] C57BL / 6J mice were selected and epileptic seizures were induced by pentylenetetrazol (PTZ) injection. Epileptic seizure behaviors and neural electrical activities were recorded synchronously by video and electroencephalography (EEG), and immunohistochemistry was used to verify the pathological changes in brain tissue after epilepsy induction.

[0012] (3) Experimental groups:

[0013] The mice were randomly divided into an experimental group (receiving focused ultrasound stimulation) and a control group (receiving sham stimulation).

[0014] (4) Stimulation parameters:

[0015] Focused ultrasound frequency range is 0.5MHz to 3MHz, power range is 0.1W / cm 2 Up to 1W / cm 2 , with pulse repetition frequencies ranging from 1 Hz to 100 Hz. Single stimulation lasted 5 minutes, once a day for 7 consecutive days, and focused ultrasound was applied to the stimulation area at different depths to evaluate depth-related effects.

[0016] (5) Data collection:

[0017] The frequency and amplitude of epileptiform discharges are recorded by EEG, calcium imaging technology is used to evaluate changes in neuronal calcium signals, PET-CT or fMRI is used to detect metabolic levels in epilepsy-related brain areas, and video is used to analyze the frequency, duration, and intensity of epileptic seizure behavior.

[0018] (6) Data analysis:

[0019] It includes behavioral analysis, neural activity analysis, brain metabolism analysis and long-term effect evaluation, statistics of epileptic seizure frequency and duration, analysis of spectral changes in EEG signals and differences in calcium signals and metabolic levels.

[0020] (7) Multimodal Verification:

[0021] Ultrasonic stimulation was applied to different brain regions (such as the hippocampus and cortex) to verify the effects of stimulation intensity and frequency on epileptic activity and screen the optimal parameter combination. Ultrasonic stimulation was also applied at different stages of the epilepsy model (acute stage and chronic stage) to analyze the differences in time effectiveness.

[0022] (8) Experimental results show that:

[0023] Focused ultrasound stimulation significantly reduces the frequency and duration of epileptic seizures, and significantly reduces abnormal neuronal discharge activity, as evidenced by a reduction in epileptiform waveforms in EEG signals. Calcium imaging shows that the spatial and temporal consistency of neuronal activity is enhanced, and abnormally active areas are significantly reduced. Brain metabolic levels tend to normalize in epilepsy-related brain areas, as evidenced by a significant recovery of glucose metabolism and blood flow. Long-term observations have shown that the effect of ultrasound stimulation is persistent and can reduce the recurrence rate of epilepsy. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a schematic diagram of the structure of a head-mounted focused ultrasound device;

[0021] Figure 2 Schematic diagram of experimental results and analysis. DETAILED DESCRIPTION

[0022] The specific implementation of the present invention is as follows: a micro focused ultrasonic transducer is used to precisely adjust the frequency (0.5 MHz to 3 MHz), power (0.1 W / cm 2 Up to 1W / cm 2 ) and pulse repetition frequency (1Hz to 100Hz) to achieve non-invasive stimulation of specific brain regions in mice. A mouse epilepsy model was established using pentylenetetrazol injection, and the target brain region was stimulated daily for 7 days using a head-mounted device. EEG signals, calcium imaging data, and brain metabolic imaging results were collected simultaneously to comprehensively analyze changes in epileptic seizure frequency, neuronal activity, and brain metabolic levels. The experimental results show that this method is highly efficient, repeatable, and has a significant effect on alleviating epilepsy, providing a new direction for non-invasive epilepsy treatment technology.

Claims

1. A method for analyzing the effect of head-mounted focused ultrasound on epileptic discharges in mice, characterized in that: It includes the design of a head-mounted focused ultrasound device and the analysis of its impact on mouse epilepsy, specifically including the design of a head-mounted focused ultrasound device, the preparation of an epileptic mouse model, the selection of focused ultrasound stimulation parameters, the synchronous experimental recording process, and the mechanism of action of focused ultrasound.

2. The design of the head-mounted focused ultrasound device according to claim 1 is characterized in that: It adopts a modular design and consists of a miniature focused ultrasound transducer, a lightweight mechanical bracket, an adjustable fixture, a cooling system and a control module. The cooling system and transducer are integrated into one to ensure the stability of long-term stimulation. The fixture can be fine-tuned according to the size of the mouse's head, with accurate positioning to avoid damage. The user interface can be used to adjust parameters in real time and monitor the status of the device. It has the function of storing and exporting experimental data to meet the needs of multiple repeated experiments.

3. The preparation of the epilepsy mouse model according to claim 1, characterized in that: Epileptic seizures were induced by pentylenetetrazol injection, and epileptic behavioral characteristics were monitored synchronously by video. Epileptic discharge activities were recorded and analyzed by electroencephalography to provide baseline data support for the intervention effect of focused ultrasound.

4. The focused ultrasound stimulation parameter selection according to claim 1, characterized in that: Focused ultrasound stimulation parameters include frequency range 0.5 MHz to 3 MHz and power density range 0.1 W / cm 2 Up to 1W / cm 2 The pulse repetition frequency range is 1Hz to 100Hz. By adjusting the fixed components of the head-mounted device, precise stimulation of the target brain area can be achieved. A head-mounted focused ultrasound device was used for a 7-day intervention experiment, with a single stimulation lasting 5 minutes per day. Different stimulation parameter combinations were combined to verify the control effect of the optimal parameters on epileptic activity, provide a basis for intervention at different stages of the disease, and optimize the intervention effect.

5. The synchronous experiment recording process according to claim 1, characterized in that: Changes in epilepsy-related brain regions were assessed using a comprehensive multimodal approach, including behavioral analysis, calcium imaging, brain metabolic imaging, and spectral analysis of EEG signals, to comprehensively and synchronously record the effects of focused ultrasound.

6. The mechanism of action of focused ultrasound according to claim 1, characterized in that: By analyzing behavioral and electrophysiological data, the frequency, duration and intensity of epileptic seizures were analyzed, and combined with brain tissue pathology analysis, the mechanism of action and efficacy of focused ultrasound were comprehensively evaluated.

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