Method for improving excitability of neurons and application thereof

Abstract

The invention provides a method for improving excitability of neurons by using terahertz waves and application thereof, and the method comprises the following steps: the neurons are radiated by using terahertz waves, the radiation frequency of the terahertz waves is 0.15-0.16 THz, and the power density of the terahertz waves is 6-13 milliwatts per square centimeter. The method disclosed by the invention can be used for improving the excitability of the neuronal cells, and can be used for preparing animal in-vitro brain slices with excited neurons or culturing to obtain the neuronal cells with improved excitability, so that the neuronal cells can be further used for scientific research and drug development.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to a method for improving neuron excitability by utilizing terahertz waves and an application thereof. Background Art

[0002] Terahertz (THz) waves are a band of electromagnetic waves with frequencies between infrared and microwaves, and their frequency range is approximately between 0.1-10 THz. Terahertz has many unique characteristics, such as non-ionizing radiation, high penetration, and specific absorption of many substances. In the body, terahertz waves can affect their structure and function by resonating with the vibration modes of biomacromolecules. In addition, terahertz waves can also change the ion concentration in cells, regulate cell membrane permeability, and affect cell metabolic activity and signal transduction. Terahertz waves under certain conditions can affect multiple organs and systems of organisms, and the brain is an important target organ of terahertz waves. However, there is a lack of research on the regulation of neural function by terahertz waves, and there are no reports on the effect on the excitability of hippocampal neurons.

[0003] Therefore, it is urgent to further explore the effects of terahertz waves on the excitability of hippocampal neurons and to develop terahertz wave exposure conditions and methods that affect the excitability of hippocampal neurons. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention provides a method for increasing neuronal excitability by using terahertz waves and its application, which can increase the excitability of neuronal cells. The method of the present invention can be used to prepare isolated brain slices of neuron-excitable animals or culture neuronal cells with increased excitability, so as to be further used in scientific research and drug development.

[0005] Therefore, in the first aspect of the present invention, the present invention provides a method for improving neuronal excitability. The method comprises:

[0006] The neurons are irradiated with terahertz waves, wherein the radiation frequency of the terahertz waves is 0.15 to 0.16 THz and the power density is 6 to 13 milliwatts per square centimeter.

[0007] After a lot of experiments and explorations, the inventors were pleasantly surprised to find that within this range of terahertz wave exposure dose, neuronal excitability is high.

[0008] According to an embodiment of the present invention, the radiation frequency of the terahertz wave is 0.152 THz.

[0009] According to an embodiment of the present invention, the power density is 12.6 milliwatts per square centimeter. At this terahertz wave exposure dose, the excitability of neurons is high.

[0010] According to an embodiment of the present invention, the power density is 6.8 milliwatts per square centimeter. Under this terahertz wave exposure dose, the excitability of neurons is high.

[0011] According to an embodiment of the present invention, the average power density of the terahertz wave is 9 to 10 milliwatts per square centimeter. Under this terahertz wave exposure dose, the excitability of neurons can be stably increased.

[0012] According to an embodiment of the present invention, the average power density of the terahertz wave is 9.7 milliwatts per square centimeter. At this terahertz wave exposure dose, neuronal excitability is significantly increased.

[0013] According to an embodiment of the present invention, the duration of the radiation treatment is 40 to 80 minutes. The inventors have obtained this optimal radiation treatment duration through a large number of experiments and explorations.

[0014] According to an embodiment of the present invention, the duration of the radiation treatment is 60 minutes. The inventor has determined this optimal radiation treatment duration through a large number of experiments.

[0015] According to an embodiment of the present invention, the neurons are hippocampal neurons. Thus, hippocampal neurons with significantly improved excitability are obtained.

[0016] According to an embodiment of the present invention, the neuron excitability includes at least one of the following: spontaneous discharge frequency, action potential, synchronous discharge frequency and neuron synaptic plasticity. The increase in spontaneous discharge frequency, action potential, synchronous discharge frequency and neuron synaptic plasticity is a sign of increased neuron excitability. The neuron treated by the method according to an embodiment of the present invention has increased spontaneous discharge frequency, action potential, synchronous discharge frequency and neuron synaptic plasticity.

[0017] In a second aspect of the present invention, the present invention provides a method for preparing an isolated brain slice of an animal. The method comprises:

[0018] Terahertz waves are used to irradiate neurons of non-human mammals, wherein the radiation frequency of the terahertz waves is 0.15-0.16 THz and the power density is 6-13 milliwatts per square centimeter.

[0019] Thus, the non-human mammal in vitro brain slices with improved neuronal excitability can be further used for scientific research and drug development. It should be noted that, in this article, the term "scientific research" can be, for example, electrophysiological research, pathophysiological research, biochemical research, pharmacological research, molecular biology research and clinical medicine research, etc. In this article, the term "drug development" can be, for example, drug discovery, drug screening, pharmacology and toxicology research, drug preparation research, drug quality research, drug safety evaluation, etc.

[0020] According to an embodiment of the present invention, the radiation frequency of the terahertz wave is 0.152 THz, the average power density is 9.7 milliwatts per square centimeter, the duration of the radiation treatment is 60 minutes, and the neurons are hippocampal neurons. As a result, the excitability of hippocampal neurons in animal brain slices in vitro is significantly improved.

[0021] According to an embodiment of the present invention, the animal in vitro brain slice has at least one of the following characteristics: enhanced spontaneous discharge frequency, enhanced action potential, enhanced synchronous discharge frequency and enhanced neuronal synaptic plasticity.

[0022] According to an embodiment of the present invention, during the process of irradiating an animal in vitro brain slice containing non-human mammalian neurons with terahertz waves, oxygenated cerebrospinal fluid is continuously perfused.

[0023] According to an embodiment of the present invention, the temperature rise of the animal's in vitro brain slices does not exceed 0.5 degrees Celsius before and after the radiation treatment.

[0024] In some embodiments of the present invention, the non-human mammal is a rodent or a primate; preferably, the non-human mammal is a mouse, a rat, a rabbit or a monkey.

[0025] In the third aspect of the present invention, the present invention proposes a use of an isolated brain slice of a non-human mammal prepared according to the aforementioned method, wherein the isolated brain slice is used for research related to neuronal excitability or for screening or identifying substances that can reduce neuronal excitability.

[0026] It should be noted that the above-mentioned uses of screening or identifying substances that can reduce neuronal excitability are non-diagnostic and non-therapeutic.

[0027] In this article, the term "neuronal excitability related research" can be, for example, related electrophysiological research, pathophysiological research, biochemical research, pharmacological research, molecular biological research and clinical medicine research.

[0028] Those skilled in the art will appreciate that the features and advantages described above for the method of increasing neuronal excitability and the method of preparing isolated animal brain slices are also applicable to this purpose and will not be described in detail here.

[0029] In a fourth aspect of the present invention, the present invention provides a method for culturing neurons. The method comprises:

[0030] The neurons are irradiated with terahertz waves, wherein the terahertz waves have a radiation frequency of 0.15 to 0.16 THz and a power density of 6 to 13 milliwatts per square centimeter, thereby cultivating neurons with significantly improved excitability.

[0031] According to an embodiment of the present invention, the radiation frequency of the terahertz wave is 0.152 THz, the average power density is 9.7 milliwatts per square centimeter, the duration of the radiation treatment is 60 minutes, and the neurons are hippocampal neurons. Thus, hippocampal neurons with significantly improved excitability are cultured.

[0032] Those skilled in the art will appreciate that the features and advantages described above for the method for increasing neuronal excitability and the method for preparing isolated brain slices of neuron-excited animals are also applicable to this method and will not be described in detail here.

[0033] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0035] Figure 1 The results of the 0.152 THz exposure dose detection and temperature monitoring of Group T in Example 1 of the present invention are shown in FIG. A is a thermal map of the dose distribution at 0.152 THz; B is a temperature change after 60 minutes of exposure at 0.152 THz;

[0036] Figure 2 The results of the investigation of the effect of terahertz waves on the spontaneous discharge of neurons in the CA1 subregion of the ventral hippocampus in Example 2 of the present invention are shown in Figure 1, wherein: A is a representative trajectory diagram of the spontaneous discharge of neurons in the CA1 subregion of the ventral hippocampus, with a scale of 500 milliseconds; B is a statistical analysis result diagram of the spontaneous discharge of neurons in the CA1 subregion of the ventral hippocampus, with * indicating p < 0.05;

[0037] FIG3 is a graph showing the effect of terahertz waves on the action potential of neurons in the CA1 subregion of the ventral hippocampus in Example 2 of the present invention, wherein: Figure 3-A Representative traces of action potentials of neurons in the CA1 subfield of the ventral hippocampus, scale bar = 25 mV or 100 ms; Figure 3-B This is the statistical result diagram of the relationship between the number of action potentials and the step pulse current, * indicates P < 0.05, ** indicates P < 0.01; Figure 3-C Representative traces of single action potentials evoked by base-strength current, scale bar = 10 mV or 1 ms; Figure 3-D is a line graph of membrane potential and dV / dt; Figure 3-E Statistical analysis of the base strength current of neurons, * indicates P < 0.05; Figure 3-F This is the statistical analysis result of action potential threshold of neurons, * indicates P < 0.05; Figure 3-GThis is the result of statistical analysis of action potential amplitude of neurons; Figure 3-H This is the statistical analysis result of the action potential half-width of neurons; Figure 3-I This is the statistical analysis result of action potential latency of neurons;

[0038] Figure 4 shows the results of the investigation of the effect of terahertz waves on the synchronous discharge activity of the CA1 subregion of the ventral hippocampus, where: Figure 4-A This is the heat map of the discharge amplitude distribution of the brain slices in group C, scale bar = 0.5 mm; Figure 4-B This is the raster image of the discharge activity of the brain slice in group C; Figure 4-C This is the heat map of the discharge amplitude distribution of the brain slice in group T, scale bar = 0.5 mm; Figure 4-D This is the raster image of the discharge activity of the brain slice in group T; Figure 4-E For statistical analysis of burst discharge frequency, * indicates P < 0.05; Figure 4-F Statistical analysis of the proportion of action potentials in burst discharges, * indicates P < 0.05;

[0039] Figure 5 shows the effect of terahertz waves on long-term potentiation of CA3-CA1 in the ventral hippocampus of mice, where: Figure 5-A Representative traces of fEPSPs after basal stimulation (dashed line) and high-frequency stimulation (solid line), scale bar = 0.5 mV or 5 ms; Figure 5-B 5-A is a schematic diagram of the change of fEPSP slope; 5-C is the statistical analysis of fEPSP slope, **** indicates P < 0.0001. DETAILED DESCRIPTION

[0040] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0041] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

[0042] Terms and Definitions

[0043] In this article, the term "hertz" is abbreviated as Hz, which is a physical unit of measurement for frequency, referring to the number of periodic fluctuations that occur per second. The main units of measurement for high frequencies are kHz (i.e. 1000Hz), MHz (i.e. 1000kHz), GHz (i.e. 1000MHz), and THz (i.e. 1000GHz). In this article, the term "terahertz" is equivalent to "THz".

[0044] As used herein, the term "brain slice in vitro" is a type of biopsy, and refers to a biopsy with a thickness of several hundred micrometers prepared from animal brain tissue.

[0045] In this article, the term "neuronal excitability" is equivalent to "neuronal excitation." The generation of action potential is an important sign of neuronal excitation.

[0046] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be purchased commercially.

[0047] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

[0048] Example 1: Terahertz wave exposure of mouse brain slices

[0049] Acute brain slices of the ventral hippocampus of mice were prepared and randomly divided into group C and group T. The slices were incubated in artificial cerebrospinal fluid at 30 degrees Celsius for 30 minutes and then incubated at room temperature for 30 minutes before use in the experiment.

[0050] Main equipment and operating instructions for terahertz exposure: The QS2-180 THz wave emission source was stabilized for 30 minutes after being turned on, and the frequency was set to 0.152 THz. The power meter measured the average power density of this frequency to be 9.7 mW / cm2, and the spot diameter was 36 mm.

[0051] Treatment method for brain slices in group T: 60 minutes of terahertz wave exposure, continuous perfusion of oxygenated cerebrospinal fluid during the process; the temperature rise of the brain slices before and after terahertz wave exposure was 0.484 degrees Celsius. The terahertz wave exposure frequency was 0.152THz, and the average power density was 9.7 mW / cm2 (single detection THz exposure dose: 6.8-12.6 mW / cm2).

[0052] In this embodiment, the temperature rise of the brain slice before and after the terahertz wave exposure is: the temperature of the brain slice after the terahertz wave exposure - the temperature of the brain slice before the terahertz wave exposure.

[0053] Treatment of brain slices in group C was the same as that in group T, but without terahertz wave exposure.

[0054] The results are as follows Figure 1 The thermal map of the terahertz wave exposure dose distribution with a radiation frequency of 0.152 THz is shown in Figure 1 A, 0.152THz exposure for 60 minutes, temperature change Figure 1 B.

[0055] Example 2: 60-minute exposure to 0.152 THz / 9.7 mW / cm2 can increase the spontaneous discharge frequency and action potential of hippocampal neurons

[0056] The brain slices of group T of Example 1 were exposed to terahertz waves according to the method of Example 1, then transferred to the recording tank and fixed with a cover net, and cells with good morphology were selected. The liquid junction potential was zeroed after the electrode was inserted. When the tip of the electrode approached the cell membrane, it could be seen that the tissue fragments on the cell surface were blown away by the spray solution wave flowing out from the tip of the pipette. Continue to move forward or lower the electrode until a small pit is formed on the cell membrane. Immediately open the valve to release the positive pressure. At this time, the square wave current pulse of "Seal Test" on the computer screen is reduced to 1 / 3-1 / 2, and the resistance is increased to GΩ, forming a good seal. Use the Gap-free recording mode, and start recording extracellular spontaneous action potentials after the cells are stable.

[0057] After the extracellular spontaneous discharge recording was completed, a short and strong negative pressure was applied to the cell using a syringe to break the cell membrane and form a whole-cell recording for whole-cell capacitance and series resistance compensation. The stimulation current was varied from 0 to 200 pA in 10 pA steps with a duration of 500 ms to record action potentials.

[0058] The results of the investigation can be found in Figure 2 , Figure 3. Among them, the representative trajectory of spontaneous discharge of neurons in the ventral hippocampal CA1 subregion is shown in Figure 2 A, Statistical analysis results of spontaneous discharges of neurons in the CA1 subregion of the ventral hippocampus are shown in Figure 2B. The firing of action potentials induced by step pulse current in neurons in the CA1 subregion of the ventral hippocampus is shown in Figure 3A; the number of action potentials fired in groups C and T under current stimulation of 150 pA and 200 pA is shown in Figure 3B; the analysis results of single action potentials induced by base current are shown in Figure 3C; the line graph of membrane potential and dV / dt is shown in Figure 3D; the change of base current after terahertz wave exposure is shown in Figure 3E; the action potential threshold of neurons is shown in Figure 3F; the action potential amplitude of neurons is shown in Figure 3G; the half-width of action potential of neurons is shown in Figure 3F; the action potential latency of neurons is shown in Figure 3H and Figure 3I.

[0059] like Figure 2 As shown, after terahertz wave exposure, the frequency of spontaneous discharges of neurons in the CA1 subregion of the ventral hippocampus recorded by attached patch clamp increased.

[0060] As shown in Figure 3, under current stimulation of 150 pA and 200 pA, the number of action potentials in group T increased; after terahertz wave exposure, the base current decreased (P < 0.05), the action potential threshold decreased (P < 0.05), and there was no significant change in the action potential amplitude, half-width and latency.

[0061] The above results show that terahertz wave radiation exposure frequency of 0.152 THz, average power density of 9.7 mW / cm2, and duration of 60 minutes can increase the spontaneous discharge frequency and action potential of hippocampal neurons, indicating increased neuronal excitability.

[0062] Example 3: 60 minutes of exposure to 0.152THz / 9.7mW / cm2 terahertz waves can increase the frequency of synchronous discharges of ventral hippocampal neurons

[0063] Use dissecting forceps to free the target brain tissue, transfer the brain tissue to the center of the Maxone chip, and gently press the brain tissue with a cover net to ensure that the position of the brain slice is fixed during the recording process. The radiation frequency is 0.152THz, the average power density is 9.7 mW / cm2, the terahertz wave is exposed for 60 minutes, and the perfusion is continued for 10 minutes during the exposure process. After the brain slice is stable, the electrical signal is recorded by MaxLabLive Scope software with a sampling rate of 20,000 Hz. The spontaneous activity of neurons is measured using Activity Scan Assay. The entire chip area is scanned by Sparse 7× sparsely (a total of 7 configurations are scanned, each configuration is 30s). The system automatically identifies active neurons based on the discharge frequency and amplitude of the scanned activity. Select the active neurons in the target area, and use Network Assay to record the neural network activity for 5 minutes. Activity Scan Assay sets the detection threshold: 0.1 Hz (discharge frequency); 20 microvolts (amplitude); 200 milliseconds (action potential interval). The threshold parameters for neural network and action potential recording were as follows: 0.3 s (smoothing window size); 1.2 (burst discharge detection threshold); 1 s (minimum peak distance); 0.3 (start and end thresholds).

[0064] The results are shown in Figure 4.

[0065] As shown in Figure 4A-D, the results of the investigation of the synchronous discharge activity of vCA1 neurons showed that the number of actively discharging electrodes increased after terahertz exposure; quantitative analysis of the synchronous discharge activity indicators of brain slices showed that compared with group C, the frequency of burst discharges in group T increased (P < 0.05) (Figure 4E), and the proportion of action potentials in burst discharges increased (P < 0.05) (Figure 4F).

[0066] The above results show that terahertz wave radiation exposure frequency of 0.152THz, average power density of 9.7mW / cm2, and duration of 60 minutes can lead to an increase in the synchronous discharge frequency of ventral hippocampal neurons.

[0067] Example 4: 60 minutes of exposure to 0.152 THz / 9.7 mW / cm2 terahertz waves can enhance synaptic plasticity in ventral hippocampal neurons

[0068] Pull a glass electrode with a tip resistance of 1-2MΩ and fill the electrode with artificial cerebrospinal fluid. Transfer the brain slice to the recording tank. Use a brush to adjust the position of the brain slice and fix it with a cover mesh. Perfuse oxygenated artificial cerebrospinal fluid at a rate of 2-4 ml / min for at least 20 minutes to stabilize the brain slice before recording. Insert the recording electrode into the stratum radiatum of the CA1 subregion of the hippocampus on the ventral side of the brain slice, and insert the stimulating electrode into the Schaeffer collateral branch of the stratum radiatum of the hippocampus, and insert it a little lower after contacting the brain slice. Adjust the electrode depth, stimulator output intensity, and the position of the two electrodes to obtain a large amplitude of field excitatory postsynaptic potentials (fEPSP), reduce the stimulation intensity to half of the maximum fEPSP, and record a 20-minute baseline. Give high-frequency stimulation to induce long-term potentiation. Record fEPSP for 30 minutes after high-frequency stimulation. The criteria for successful induction of long-term potentiation: fEPSP amplitude or slope increases by ≥30% and is maintained for ≥30 minutes.

[0069] The results are shown in Figure 5. The fEPSP trajectories of each group after basal stimulation and high-frequency stimulation are shown in Figure 5A. After high-frequency stimulation, the fEPSP slope of group C slowly decreased, and the fEPSP slope of group T did not decrease significantly within 30 minutes, as shown in Figure 5B. The quantitative analysis results showed that compared with group C, the average fEPSP slope of group T at 40-50 minutes was significantly increased, as shown in Figure 5C.

[0070] The above results show that terahertz wave radiation exposure frequency of 0.152THz, average power density of 9.7mW / cm2 and duration of 60 minutes can enhance the synaptic plasticity of ventral hippocampal neurons.

[0071] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, unless they are contradictory.

[0072] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for increasing neuronal excitability, characterized in that: include: The neurons are irradiated with terahertz waves, wherein the radiation frequency of the terahertz waves is 0.15 to 0.16 THz and the power density is 6 to 13 milliwatts per square centimeter.

2. The method according to claim 1, characterized in that The radiation frequency of the terahertz wave is 0.152 THz; Optionally, the power density is 12.6 milliwatts per square centimeter; Optionally, the power density is 6.8 milliwatts per square centimeter; Optionally, the average power density of the terahertz wave is 9 to 10 milliwatts per square centimeter; Optionally, the average power density of the terahertz wave is 9.7 milliwatts per square centimeter.

3. The method according to claim 1, characterized in that The duration of the radiation treatment is 40 to 80 minutes.

4. The method according to claim 3, characterized in that: The duration of the radiation treatment was 60 minutes.

5. The method according to claim 1, characterized in that The neurons are hippocampal neurons.

6. The method according to any one of claims 1 to 5, characterized in that: The neuronal excitability includes at least one of the following: spontaneous discharge frequency, action potential, synchronous discharge frequency and neuronal synaptic plasticity.

7. A method for preparing animal brain slices in vitro, characterized in that: include: Terahertz waves are used to radiate animal in vitro brain slices containing non-human mammal neurons. The radiation frequency of the terahertz waves is 0.15-0.16 THz and the power density is 6-13 milliwatts per square centimeter.

8. The method according to claim 7, characterized in that The radiation frequency of the terahertz wave is 0.152 THz, the average power density is 9.7 milliwatts per square centimeter, and the duration of the radiation treatment is 60 minutes; the neurons are hippocampal neurons; Optionally, the animal ex vivo brain slice has at least one of the following characteristics: enhanced spontaneous discharge frequency, enhanced action potential, enhanced synchronous discharge frequency and enhanced neuronal synaptic plasticity; Optionally, the temperature of the animal's ex vivo brain slices rises by no more than 0.5 degrees Celsius before and after the radiation treatment; Optionally, the non-human mammal is a rodent or a primate; preferably, the non-human mammal is a mouse, a rat, a rabbit or a monkey.

9. Use of non-human mammal in vitro brain slices prepared according to the method of claim 7 or 8, characterized in that: The isolated brain slice is used for research related to neuronal excitability or for screening or identifying substances that can reduce neuronal excitability.

10. A method for culturing neurons, characterized in that: include: The neurons are irradiated with terahertz waves, wherein the terahertz waves have a radiation frequency of 0.15 to 0.16 THz and a power density of 6 to 13 milliwatts per square centimeter; Optionally, the radiation frequency of the terahertz wave is 0.152 THz, the average power density is 9.7 milliwatts per square centimeter, the duration of the radiation treatment is 60 minutes; and the neurons are hippocampal neurons.

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