Terahertz regulation and control method for improving mitochondrial activity of nerve cells and application

Abstract

The invention discloses a terahertz regulation and control method for improving mitochondrial activity of nerve cells and application, and belongs to the technical field of terahertz. The method disclosed by the invention comprises the step of performing scanning type irradiation on the nerve cells by using terahertz waves with the irradiation frequency of 27.27-59.41 THz as a light source. The method comprises the following step: scanning and irradiating nerve cells by using terahertz waves with the light frequency of 34.9 THz, the light power density of 3.98 W / cm < 2 > and the irradiation time of 43 minutes. According to the present invention, the ATP content of the irradiated cells is detected, such that the ATP content of the nerve cells is increased, the mitochondrial metabolism is promoted, and the mitochondrial activity is improved under the irradiation condition, and the terahertz wave can improve the mitochondrial activity of the cells by enhancing the ATP synthesis ability of the nerve cells.

Description

Technical Field

[0001] The present invention belongs to the field of terahertz technology, and specifically relates to a terahertz regulation method for increasing the activity of nerve cell mitochondria, especially energy metabolism to produce ATP. Background Art

[0002] Mitochondria are the metabolic center of cellular energy and important organelles of eukaryotic cells. They generate energy through oxidative phosphorylation and regulate cell metabolic homeostasis, proliferation / differentiation, and cell death. The normal activity of mitochondria is closely related to the normal functioning of cell functions. And its key role is that the disorder of energy metabolism will cause abnormalities in cell structure and function, leading to cell death. Nerve cells are extremely sensitive to energy changes, and their energy metabolism is extremely vigorous. Therefore, mitochondria play an important role in major diseases such as aging and neurodegenerative diseases.

[0003] Terahertz waves are high-frequency electromagnetic waves that have great scientific exploration value and potential application value due to their spectrum resolution, penetration, safety and unique biological effects. Since the 1980s, terahertz technology has begun to develop rapidly and has been applied in many fields. Since the terahertz spectrum is close to the energy level of many molecular vibrations and rotations and weak interactions between molecules, it can interact with many cells, tissues and biomacromolecules and cause a series of biological effects, so it is believed to have the function of regulating biological organisms.

[0004] In neurological diseases, changes in mitochondrial structure and function are inevitable. Existing studies mostly use drugs for treatment and regulation. In a complex cellular environment, the targeting and efficacy of drugs will be greatly reduced. The use of light regulation technology can target the target location, which is simpler and more effective for trauma patients. However, due to the late development of terahertz technology, existing electromagnetic radiation research on nerve cells is mostly concentrated on visible light, near-infrared light and microwaves. In these existing related studies, most of them rely on the thermal effect of electromagnetic radiation for activity regulation. The low-energy characteristics of terahertz waves themselves rely on non-thermal effects for regulation, thereby avoiding adverse reactions caused by cellular thermal effects, especially sensitive nerve cells. However, there is currently a lack of relevant neurobiological research in the terahertz band, especially the regulation of mitochondrial activity by terahertz waves. Summary of the invention

[0005] In view of the problems existing in the above-mentioned prior art, the purpose of the present invention is to design and provide a terahertz regulation method for improving mitochondrial activity, targeting the ATP synthase located in the inner membrane of mitochondria responsible for generating ATP in the cell, and performing specific regulation by means of terahertz technology, thereby increasing ATP generation, promoting mitochondrial metabolism, and improving mitochondrial activity.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] On the one hand, the present invention provides a terahertz regulation method for improving the mitochondrial activity of nerve cells, comprising using a terahertz wave with an irradiation frequency of 27.27 to 59.41 THz as a light source to scan and irradiate nerve cells. After irradiation, the change in ATP content in the nerve cells after irradiation is measured.

[0008] In the terahertz control method for improving the mitochondrial activity of nerve cells, the irradiation frequency is 34.9 THz. By detecting the change in the ATP content of the cells after light irradiation at the light frequency, it is convenient to determine whether the mitochondrial activity in the nerve cells is increased.

[0009] In the terahertz control method for improving the activity of nerve cell mitochondria, the improvement of nerve cell mitochondrial activity is to increase the ATP content. The increase in ATP content is an indication of the change in the activity of nerve cell mitochondria.

[0010] The terahertz control method for improving the mitochondrial activity of nerve cells is characterized in that the power density of the terahertz wave is 3.6 to 4.2 W / cm 2 .

[0011] The terahertz control method for improving the activity of nerve cell mitochondria, the power density of the terahertz wave is 3.98W / cm 2 .

[0012] In the terahertz control method for improving the mitochondrial activity of nerve cells, the irradiation time of the terahertz wave is 40 to 45 minutes.

[0013] In a second aspect, the present invention provides the use of any of the methods described above in the preparation of a health product for improving the mitochondrial activity of nerve cells.

[0014] In a third aspect, the present invention provides use of any of the methods described in any one of the above in the preparation of a drug for treating aging or neurodegenerative diseases.

[0015] Among them, the terahertz wave with a frequency of 34.9THz is obtained by molecular dynamics simulation to calculate the infrared absorption spectrum of ATP synthase located in the inner membrane of mitochondria. ATP synthase in the inner membrane of mitochondria is a type of adenosine triphosphatase (ATPase), specifically F-type FoF1-ATP synthase (F Type FoF1 ATP Synthase), which is located on the inner membrane of mitochondria and uses the proton motive force generated by the oxidative respiratory chain to synthesize adenosine triphosphate (ATP) by causing conformational changes in proteins. Most of the ATP in cells comes from ATP synthase.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The terahertz regulation method for improving the activity of nerve cell mitochondria of the present invention can enhance the increase of ATP content in nerve cells under the light energy of 34.9THz, thereby improving the activity of nerve cell mitochondria and enhancing energy metabolism, which has certain significance for treating diseases related to mitochondrial energy metabolism. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 is the result of molecular dynamics simulation, where: A is a simplified schematic diagram of the proton transport region of ATP synthase constructed by molecular dynamics simulation, B is the infrared absorption spectrum of the amino acid side chain at the entrance of the proton channel under different protonation states calculated by molecular dynamics simulation, the horizontal axis is the frequency in terahertz frequency, and the vertical axis is the relative intensity of the absorbance;

[0019] Figure 2 It is a simplified diagram of the light irradiation system, where A is a simplified diagram of the light irradiation system and B is the scanning process;

[0020] Figure 3 The detection results of ATP content, wherein A is the fluorescence imaging of the control group, B is the fluorescence imaging of the irradiated group, C is the statistical result of red fluorescence in Figures A and B, the ordinate is the relative fluorescence intensity, the average value of the control group is set to 1, and ** indicates p < 0.01 compared with the control group. DETAILED DESCRIPTION

[0021] The specific implementation of the present invention is described below in conjunction with the accompanying drawings so that those skilled in the art can better understand the present invention. It should be noted that in the following description, when the detailed description of known functions and designs may dilute the main content of the present invention, these descriptions will be omitted here.

[0022] The terahertz regulation method of the present invention for improving the mitochondrial activity of nerve cells is to use terahertz waves with a light frequency of 34.9THz to scan and irradiate the cells, realize terahertz regulation of the cells in the irradiated area, increase the synthesis of cell ATP, and thus improve mitochondrial activity. The present invention first finds through molecular dynamics simulation modeling calculation that the carboxyl group of the key amino acid side chain at the entrance of the proton transport channel of the key protein ATP synthase for ATP generation shows an obvious infrared absorption characteristic peak at 34.9THz after binding to protons, which means that the ATP synthase in the transport proton has the potential to absorb light with a light frequency of 34.9THz, thereby affecting its function. Using specific ATP fluorescent dyes, staining and detection of nerve cell lines irradiated with light with a light frequency of 34.9THz are carried out, and it is found that the ATP generation of cells after irradiation is significantly increased, that is, the activity of mitochondria is significantly enhanced.

[0023] Embodiment 1:

[0024] Through molecular dynamics simulation calculation, according to the infrared absorption spectrum results, the light frequency absorbed by ATP synthase is obtained. The specific process is as follows:

[0025] The infrared absorption spectrum is calculated for ATP synthase. ATP synthase, located in the inner membrane of mitochondria, is generated by the existence of the proton gradient difference between the inner and outer membranes and is the main source of cellular energy. Its structure is complex, and the Fo part is the proton transport region. The currently believed proton transport process is to transport protons through the c8 ring and a subunit of the Fo part (human ATP synthase). When the proton concentration in the mitochondrial intermembrane cavity is higher than that in the mitochondrial matrix, protons are transported from the mitochondrial intermembrane cavity to the mitochondrial matrix, driving the rotation of ATP synthase to synthesize ATP. This process of transport is mainly carried out through a series of glutamate residues (Glutamate, Glu) on the c8 ring and the a subunit for proton transport, and the positively charged arginine residues on the a subunit assist in transport. The proton binds to the carboxyl residue of glutamate at the entrance of the a subunit, enters the hemichannel of the mitochondrial intermembrane cavity, and then transfers to the carboxyl residue of glutamate in the c8 ring. After rotating through the positions of about 5 glutamate residues, it reaches and transfers to the carboxyl residue of glutamate at the exit of the a subunit, enters the mitochondrial matrix hemichannel, and finally enters the mitochondrial matrix to complete the transport process. ATP is produced during the transport process. When the substrate is sufficient, the faster the proton transport speed, the more ATP is synthesized, and the higher the mitochondrial activity. The key site in the transport process is the carboxyl side chain of the amino acid residue at the entrance of proton transport. Therefore, two models were constructed based on the different protonation states of the carboxyl side chain of the amino acid at the entrance.

[0026] The model was constructed with the help of Charmm-GUI online website. The model was optimized and deleted to a certain extent. According to the potential difference between the inside and outside of the mitochondrial inner membrane and the direction of proton transport, the protonation state of the side chain carboxyl groups of other amino acids involved in the proton transport process was changed to make the model closer to the actual situation. The schematic diagrams of the two models finally constructed are shown in the figure. Figure 1 As shown in A, except for the changes in the sites listed below, the other sites of the two models remain in the initial state. Here, the side chain that first accepts the proton in the c8 ring is defined as chain 1, and the chains passed by in clockwise rotation are 2 to 8, a total of eight chains, among which the 58-position glutamic acid residues of chains 1 to 6 are protonated, and the glutamic acid residue at the entrance of the a subunit is protonated and non-protonated in the two models respectively. The above is the relevant information of the constructed model. Figure 1In A, the gray area is the mitochondrial inner membrane, the black double circle area is the c8 ring of the ATP synthase Fo part, the blue linear range is the subunit a of the ATP synthase Fo part, the red circle indicates the protonated amino acid side chain, and the blue circle indicates the unprotonated amino acid side chain.

[0027] Then, the two models were simulated with the help of molecular dynamics simulation software. After the model was stabilized, the infrared absorption spectrum of the amino acid side chain carboxyl group at the entrance was calculated. The calculation results are as follows Figure 1 B. By analyzing the changes in the absorption peak position before and after the protonation state, it is shown that 34.9 THz is a special absorption peak that affects the structure and function of ATP synthase. The values ​​between different models cannot be compared.

[0028] Embodiment 2:

[0029] Using terahertz quantum cascade laser (DayLight) as the light source, the frequency coverage range can cover 27.27~59.41THz. The scanning irradiation process is: automatic scanning of the xy plane, the scanning system is as follows Figure 2 As shown in the schematic diagram, the entire scanning process lasts for about 43 minutes. The irradiated cells are HT22 (mouse hippocampal neuronal cell line).

[0030] Detection of ATP content in the cells after the above scanning irradiation:

[0031] Fluorescence imaging detection was performed with the help of ATP Red-Live (MCE) fluorescent probe and Hoechst 33342 (Biyuntian) to observe the changes in ATP production after cells were irradiated by terahertz. Among them, the ATP-Red Live probe specifically stains ATP, and the fluorescence imaging result is the red channel. The stronger the fluorescence intensity, the higher the ATP production; Hoechst 33342 stains the cell nucleus and is used for cell positioning. The specific detection steps are as follows:

[0032] The cells were stained in a T25 cell culture flask, and staining solution was added to make the concentration of the staining solution 5 μM (ATP-Red Live) and 1× (Hoechst 33342), respectively, and incubated in a 37°C cell culture incubator for 30 minutes; after the incubation, all cells were collected and planted in a cell culture dish (iBIDI), and terahertz wave irradiation was performed after the cells attached to the wall and grew in good morphology. After the irradiation, fluorescence imaging was performed using a fluorescence microscope (Leica). The imaging results are shown in FIG. Figure 3As shown in A and B, the red color in A and B represents the result of ATP Red Live fluorescent dye staining, indicating the ATP content; the blue color represents the result of Hoechst33342 live cell dye staining, indicating the position of the cell nucleus. After staining, the red fluorescence intensity at the corresponding position of the image was counted with the help of Image J. The average fluorescence intensity of the cells was counted, and the statistical results are shown in Figure 3 As shown in C, it can be seen that the relative fluorescence intensity of the irradiated group is significantly improved compared with the control group. After the present invention is adopted, the ATP of the cells, that is, the mitochondrial activity of the nerve cells, is significantly increased after the cells are irradiated with 34.9THz terahertz waves.

Claims

1. A terahertz control method for improving the mitochondrial activity of nerve cells, characterized in that: It involves using terahertz waves with a frequency of 34.9THz as a light source to scan and irradiate nerve cells.

2. A terahertz control method for improving mitochondrial activity of nerve cells according to claim 1, characterized in that: The irradiation frequency is 27.27-59.41 THz.

3. A terahertz control method for improving mitochondrial activity of nerve cells according to claim 1, characterized in that: The improving of nerve cell mitochondrial activity is to increase ATP content.

4. A terahertz control method for improving mitochondrial activity of nerve cells according to claim 1, characterized in that: The power density of the terahertz wave is 3.6-4.2 W / cm 2 .

5. The terahertz control method for improving the mitochondrial activity of nerve cells according to claim 1, characterized in that: The power density of the terahertz wave is 3.98 W / cm 2 .

6. A terahertz control method for improving mitochondrial activity of nerve cells according to claim 1, characterized in that: The terahertz wave irradiation time is 40 to 45 minutes.

7. Use of the method according to any one of claims 1 to 6 in preparing a health product for improving the mitochondrial activity of nerve cells.

8. Use of the method according to any one of claims 1 to 6 in the preparation of a drug for treating aging or neurodegenerative diseases.

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