System and product for regulating blood sugar and / or treating diabetes mellitus based on acoustic genetics and application

Abstract

The invention belongs to the technical field of ultrasonic therapy, and particularly discloses a system and a product for regulating blood sugar and / or treating diabetes mellitus based on acoustic genetics and application. The system comprises: an expression vector implantation module, which is used for implanting a vector capable of expressing a high-conductivity mechanical sensitive channel (MscL) mutant into a subject for expression; and the pulsed ultrasound instrument is used for outputting low intensity pulsed ultrasound (LIPUS) and performing LIPUS stimulation on a target area of a subject implanted with the carrier capable of expressing the MscL mutant. According to the application, acoustic genetics and an ultrasonic treatment technology are combined, and the activity of an MscL channel is regulated and controlled in a dose-dependent manner through LIPUS, so that ion balance and pressure inside and outside cells are influenced, the release amount of insulin is increased or the sensitivity of insulin is improved, accurate regulation and control of blood sugar through an acoustic dose effect are realized, symptoms of diabetes mellitus are relieved, and the risk of complications is reduced.

Description

Technical Field

[0001] The present application relates to the technical field of ultrasonic therapy, and particularly to a system, product and application for regulating blood glucose and / or treating diabetes based on optogenetics. Background Art

[0002] Type 2 Diabetes Mellitus (T2DM), as one of the most common chronic metabolic diseases globally, its incidence is closely related to multiple factors such as obesity, poor lifestyle and genetic background. Poor blood glucose control in T2DM patients can induce various complications, such as cardiovascular diseases, kidney diseases, retinopathy, neuropathy, etc., endangering the life safety of patients. Traditional treatment methods mainly include drug treatment, diet control and exercise, however, these methods have poor effects in some patients, or there are side effects and compliance problems caused by long-term drug use.

[0003] The pathogenesis of T2DM involves multiple factors, including insulin resistance, physiological processes of blood glucose regulation, and aspects such as dyslipidemia. Its pathophysiological characteristics are mainly manifested as insulin resistance and initial hyperinsulinemia. The ability of pancreatic β-cells to produce insulin gradually decreases, and the variable combination of β-cell dysfunction and insulin resistance leads to the complexity of type 2 diabetes. Low-intensity ultrasound, as a potential treatment means, has shown its role in the treatment of T2DM. Some studies have shown that insulin release induced by low-frequency, low-intensity therapeutic ultrasound can significantly increase insulin concentration in mouse models. In addition, chronic inflammation is also a common co-existing disease of T2DM. Ultrasound treatment can reduce insulin resistance and improve insulin sensitivity by inhibiting chronic inflammatory responses. Existing studies have explored the potential role of low-intensity ultrasound in inflammation regulation. Low-intensity ultrasound mainly produces mechanical (compression) effects, such as sonoporation, which can cause transient changes in cell membranes, improve cell membrane permeability, and thus affect the secretion and regulation of inflammation-related factors.

[0004] Low Intensity Pulsed Ultrasound (LIPUS), as an emerging physical therapy, is a safe and effective non-invasive treatment means that mainly relies on mechanical effects and has gradually been applied in clinical practice. In the previous experimental studies of the research group where the inventors of the present application are located, it has been proved that LIPUS can significantly improve the blood glucose level and insulin resistance of T2DM rats, which provides technical ideas and research support for the treatment of diabetes; after irradiating the liver, pancreas and other parts with LIPUS ultrasound, the metabolites in the rats generally show a significant upward trend, among which the expression of DHA increases, having a good anti-inflammatory effect, and the increase of citrate in the tricarboxylic acid model promotes energy metabolism and improves metabolic function.

[0005] However, the current research methods for treating type 2 diabetes based on LIPUS are single, and the ultrasonic parameters used cannot achieve precise regulation of blood glucose, nor is there a systematic study on ultrasonic dose. Therefore, it is of great significance to explore the combination of ultrasonic treatment technology to achieve precise regulation of ultrasonic dose-blood glucose effect and dynamically and precisely regulate blood glucose for the prevention and treatment of diabetes. Summary of the Invention

[0006] In view of the above-mentioned disadvantages of the prior art, the purpose of this application is to provide a system, product and application for regulating blood glucose and / or treating diabetes based on optogenetics, which can regulate the blood glucose level of type 2 diabetes patients by combining optogenetics and ultrasonic treatment technology, achieve precise regulation of the acoustic dose-blood glucose effect, dynamically and precisely regulate blood glucose, and provide new ideas for the prevention and treatment of diabetes.

[0007] To achieve the above object and other related objects, the first aspect of this application provides a system for regulating blood glucose and / or treating diabetes based on optogenetics, including:

[0008] An expression vector implantation module for implanting a vector capable of expressing a mutant of the Mechanosensitive Channel of Large conductance (MscL) into a subject for expression;

[0009] A pulsed ultrasound instrument for outputting LIPUS and performing LIPUS stimulation on the target area of a subject implanted with the vector capable of expressing the MscL mutant.

[0010] In some embodiments, the MscL mutant is the MscL-G22S mutant. The MscL-G22S mutant is obtained by introducing a serine (S) mutation at the 22nd amino acid of the MscL channel protein. This mutation makes the MscL channel more sensitive to low-intensity ultrasonic waves, thus playing an important role in optogenetic technology.

[0011] In some embodiments, the vector is the viral vector rAAV / pan:EGFP-pA. The viral vector rAAV / pan:EGFP-pA is a recombinant adeno-associated virus (rAAV) vector, and its core characteristics include:

[0012] rAAV: Recombinant adeno-associated virus, which is one of the most commonly used viral vectors in the field of gene therapy, and has the characteristics of high safety, low immunogenicity, wide host range, etc.;

[0013] pan: represents "pan - serotype", indicating that the vector may adopt a capsid design capable of spanning multiple AAV serotypes, or use an engineered pan - serotype capsid protein;

[0014] EGFP: Enhanced Green Fluorescent Protein, used as a reporter gene to label and track transfected cells;

[0015] pA: polyadenylation signal, ensuring the correct processing and stable expression of post - transcriptional mRNA.

[0016] In some embodiments, the vector is a pReceiver - M56 Expression Clone inserted with a fluorescent tag protein, and the fluorescent tag protein includes but is not limited to red fluorescent protein mCherry, etc. The pReceiver - M56 Expression Clone is a mammalian expression vector developed by GeneCopoeia and has the ability to highly express foreign genes.

[0017] In some embodiments, the ultrasonic dose of the LIPUS is 5 - 30 J, preferably 5 - 15 J, more preferably 11.76 J, and the calculation formula for the ultrasonic dose is: ultrasonic dose = sound intensity × area × time × duty cycle.

[0018] In some embodiments, the ultrasonic output frequency of the LIPUS is 1 MHz.

[0019] In some embodiments, the duty cycle of the LIPUS is 10% - 30%, preferably 20%.

[0020] In some embodiments, the LIPUS stimulation lasts for at least 10 days with at least one irradiation per day, and each irradiation lasts for 10 - 30 min.

[0021] In some embodiments, the LIPUS stimulation lasts for at least 10 days with one irradiation per day and each irradiation lasts for 10 min. In some embodiments, the target area is the pancreas and / or the liver.

[0022] The second aspect of the present application provides a product for regulating blood glucose and / or treating diabetes based on optogenetics, including: a vector capable of expressing an MscL mutant, and a pulsed ultrasound instrument for outputting LIPUS.

[0023] In some embodiments, the MscL mutant is the MscL - G22S mutant.

[0024] In some embodiments, the vector is the viral vector rAAV / pan:EGFP-pA.

[0025] In some embodiments, the vector is the pReceiver-M56 Expression Clone inserted with a fluorescent tag protein, and the fluorescent tag protein includes but is not limited to the mCherry tag, etc.

[0026] In some embodiments, the operating parameters of the pulsed ultrasound device include: an ultrasound dose of 5 - 30 J; and / or, an ultrasound output frequency of 1 MHz; and / or, a duty cycle of 10% - 30%; and / or, a continuous irradiation time: at least 10 days, at least once a day, and each irradiation for 10 - 30 min.

[0027] The third aspect of the present application provides the application of LIPUS and a vector capable of expressing the MscL mutant in the preparation of a product for regulating blood glucose and / or treating diabetes.

[0028] As described above, the system, product, and application for regulating blood glucose and / or treating diabetes based on optogenetics of the present application have the following beneficial effects:

[0029] The present application combines optogenetics and ultrasound therapy technologies, and uses LIPUS to dose-dependently regulate the activity of the MscL channel, thereby affecting the ion balance and pressure inside and outside the cell, increasing the insulin release amount or improving insulin sensitivity, achieving precise regulation of blood glucose by the acoustic dose effect, alleviating diabetes symptoms, and reducing the risk of complications, and has important clinical application prospects. Description of the Drawings

[0030] Figure 1 Shows the expression of mCheery in 293T cells and (A) min 6 cells (B) in the examples of the present application.

[0031] Figure 2 Shows the viability of 293T cells (A) and min6 cells in each group in the examples of the present application.

[0032] Figure 3 Shows the cell viability of min 6 cells under different ultrasound doses in the examples of the present application.

[0033] Figure 4 Shows the OGTT data (A) and AUC values (B) of each group of mice in the examples of the present application.

[0034] Figure 5 Shows the random blood glucose test results of each group of mice in the examples of the present application.

[0035] Figure 6Shown are the fasting blood glucose test results of each group of mice before and after ultrasound treatment in the embodiments of the present application.

[0036] Figure 7 Shown are the q-PCR test results of the pancreatic tissues of each group of mice in the embodiments of the present application.

[0037] Figure 8 Shown is the H&E staining result diagram of the pancreas of each group of mice in the embodiments of the present application.

[0038] Figure 9 Shown is the Oil Red O staining result diagram of the liver of each group of mice in the embodiments of the present application. Detailed implementation manners

[0039] The following uses specific specific examples to illustrate the implementation manners of the present application. Those skilled in the art can easily understand other advantages and effects of the present application from the content disclosed in this specification. The present application can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present application.

[0040] In the present application, unless otherwise specified, the term "a plurality of" means two or more.

[0041] The character " / " means that the objects before and after are in an "or" relationship. For example, A / B means: A or B.

[0042] The term "and / or" is an associative relationship describing an object, indicating that there can be three relationships. For example, A and / or B means: A or B, or, the three relationships of A and B.

[0043] As an emerging regulation technology, optogenetics has attracted much attention in recent years, and its potential in the treatment of nervous system diseases has been preliminarily verified. In this context, combining optogenetics and ultrasound treatment technology to regulate the blood glucose level of type 2 diabetes patients has important clinical application prospects.

[0044] Optogenetics combines acoustics and genetics technologies and is an emerging synthetic biology technology that precisely regulates cells using sound waves. It mainly relies on sound waves to regulate mechanosensitive ion channels (MSCs) to regulate cells to achieve precise manipulation and regulation of cells. Through molecular biology techniques, genes that can express MSCs proteins are transferred into the cells of specific tissues of the host, inducing the target cells to specifically express MSCs proteins. Then, the mechanical force generated by ultrasonic waves activates MSCs to the open state, enabling the corresponding ions inside and outside the target cells to enter the opened ion channels to form ion currents, thereby changing the membrane potential, activating or inhibiting the cells, and further regulating the cell behavior and function.

[0045] Among MS proteins with diverse structures, MscL from Escherichia coli is one of the most well-studied channels. MscL belongs to a homotetramer. The amino acid sequences of the single subunits encoded by genes of different species contain 120 - 159 amino acid sites. There are 2 transmembrane helical regions (TM1 and TM2) on each subunit. The TM1 of 5 subunits together form a pore that allows ions to pass through. MscL does not have ion selectivity. When in the open state, the pore diameter is about 2.8 nm. At this time, organic substances such as ions and small molecules with a diameter less than 2.8 nm can pass through. The main physiological function of MscL is to sense the osmotic pressure change in the surrounding environment. When the osmotic pressure suddenly decreases, the cell membrane tension changes, activating the MscL channel to release Ca 2+ 、K + and other solutes, reducing the osmotic pressure of the internal environment and preventing the cell from swelling and bursting.

[0046] This application discovers that dose-dependently regulating the activity of the MscL channel through LIPUS can affect the ion balance and pressure inside and outside the cell, increase the release amount of insulin or improve insulin sensitivity, thereby achieving precise regulation of blood glucose by acoustic dose effect, alleviating diabetic symptoms, and reducing the risk of complications.

[0047] The following is verified through cell and animal experiments. The specific implementation process is as follows:

[0048] (I) Construction of CS-GS3778-M56 plasmid

[0049] Using pReceiver-M56 Expression Clone as the vector, insert the MscL-G22S sequence and the mCherry tag. The control plasmid only inserts the mCherry tag. Entrust a biological company to construct the CS-GS3778-M56 plasmid and the control group EX-NEG-M56 plasmid.

[0050] (II) Confirm that the MscL-G22S channel can be heterologously expressed in different cells

[0051] Take HEK 293T (human embryonic kidney cells) and Min 6 (mouse pancreatic islet β cells) cells in the logarithmic growth phase, inoculate them in a 6-well plate at a density of 5×10 5 cells / mL, and culture them at 37 °C for 24 h; after the cells adhere, perform plasmid transfection, then put the cells back into the incubator and continue to culture. After 6 h, change the medium to complete medium and continue to culture. Take pictures under the microscope after 48 h as Figure 1 shown.

[0052] (III) Effect of CS-GS3778-M56 plasmid on the viability of 293T and Min 6 cells

[0053] Take 293T and Min 6 cells in the logarithmic growth phase and inoculate them into a 96-well plate at a density of 5×10 3 cells / mL and culture them at 37°C for 24 h;

[0054] After the cells adhere to the wall, perform plasmid transfection and then group the treatments. Specifically, divide them into a Blank group (i.e., the blank group without plasmid transfection), an NC group (i.e., the control group with control plasmid transfection), and an MscL group. Subsequently, replace the medium in the 96-well plate with fresh medium containing 10% CCK-8 solution, being careful to avoid generating bubbles, and place it back in the incubator for 2 h. Measure the optical density (OD) value of each well at 450 nm using an enzyme-linked immunosorbent assay reader. The higher the OD value, the stronger the cell viability. The results are as Figure 2 shown.

[0055] As Figure 2 shown, there was no difference in cell viability among the groups in 293T cells; in Min 6 cells, the cell viability in the MscL group increased (P<0.05).

[0056] (IV) Effects of different doses of LIPUS on the viability of transfected Min 6 cells

[0057] Take Min 6 cells in the logarithmic growth phase and inoculate them into a 96-well plate at a density of 5×10 3 cells / mL and culture them at 37°C for 24 h; after the cells adhere to the wall, perform plasmid transfection, and then place the cells back in the incubator for continued culture. After 6 h, replace the medium with complete medium and continue culturing. After 48 h, collect the cells. After 24 h, group them according to different ultrasound doses (ultrasound dose = intensity × area × time × duty cycle), specifically divided into an NC group (i.e., the control group transfected with plasmid but without ultrasound treatment), a 5.88J LIPUS group, an 11.76J LIPUS group, a 17.64J LIPUS group, and a 23.52J LIPUS group. Among them, the ultrasound output frequency is 1 MHz for all groups, and the duty cycle is 20% for all groups.

[0058] Subsequently, replace the medium in the 96-well plate with fresh medium containing 10% CCK-8 solution, being careful to avoid generating bubbles, and place it back in the incubator for 2 h. Measure the optical density (OD) value of each well at 450 nm using an enzyme-linked immunosorbent assay reader. The higher the OD value, the stronger the cell viability. The results are as Figure 3 shown.

[0059] As Figure 3 shown, compared with the NC group, the cell viability in the 11.76J LIPUS group increased (P<0.05).

[0060] (V) Establishment of a mouse model

[0061] Twenty male C57BKS-db / m mice and sixty C57BKS-db / db mice (n = 80) were provided by the Laboratory Animal Center of Chongqing Medical University. They were divided into a normal group, a T2DM-only group, a T2DM + virus control group (rAAV / pan:EGFP-pA, commercially available), and a T2DM + virus group (rAAV / pan:MscL-G22S-EGFP-pA, prepared by a commercial company with GMP-level production capacity, and the final product titer was 5.05×10 13 vg / mL). The normal group was fed a basal diet for routine breeding. Blood samples were collected from the tail vein of db / db mice to measure random blood glucose. When the random blood glucose was ≥16.7 mmol / L for three consecutive days, it was proved that the model was successfully established. Four weeks later, ultrasonic irradiation treatment at different doses was started.

[0062] (VI) LIPUS treatment

[0063] The hair in the target area and the surrounding area of the mice was shaved off, and the hair was completely removed with depilatory cream (Veet). The target area was determined according to the physiological anatomy of the mice, and the LIPUS detector (Chongqing Rong Hai Engineering Research Center of Ultrasound Medicine Co,Ltd,Chongqing,China) was placed on the target area, and then ultrasonic stimulation or sham stimulation could be started. When LIPUS was working, the signal was transmitted between the LIPUS transducer and the target area through an ultrasonic coupling agent (Tianjin Chengxin Medical Auxiliary Materials Factory, Tianjin, China). The sham control included all steps of LIPUS treatment, except for activating the LIPUS transducer during stimulation (the transducer power was turned off). During the treatment process, the depth of the detector relative to the animal should be kept constant at all times (the low-intensity ultrasonic transducer was kept at a fixed height above the laboratory workbench) to ensure that no pressure was exerted on the internal organs through the transducer. After the LIPUS stimulation or sham stimulation was completed, the mice were returned to their respective cages. In this experiment, a pulsed ultrasound instrument was used to irradiate the pancreas and liver of the mice with an average acoustic dose of 11.76 J for 10 minutes each time, once a day for 10 days.

[0064] (VII) Oral glucose tolerance test (OGTT)

[0065] After the mice were fasted overnight for 12 hours, the fasting blood glucose was recorded, and then a single oral administration of 2 g / kg glucose solution was given by gavage with an oral syringe. After glucose administration, blood was collected from the tail vein at four time points: 30 min, 60 min, 90 min, and 120 min to record blood glucose. After the experiment, the rats were returned to their respective cages and fed with normal feed, and the AUC-OGTT was calculated using the AUC determined by the glucose levels at baseline and 120 min after glucose overload. As Figure 4 shown, the AUC of the db / db LIPUS+MscL group decreased significantly (P<0.05).

[0066] (VIII) Blood Glucose Measurement

[0067] Whole blood samples were obtained by tail vein sampling, and a commercially available handheld blood glucose meter was used to evaluate the glucose concentration. The blood glucose instrument uses a small blood volume (0.3 μL), so no additional fluid is required to restore the total volume after blood sampling. As Figure 5 shown, the blood glucose in the db / db LIPUS group was lower than that in the db / db control group, and the blood glucose in the db / db LIPUS+MscL group was significantly lower than that in the db / db control group and the db / db LIPUS group (P<0.05).

[0068] (IX) Fasting Blood Glucose Measurement

[0069] The mice were fasted overnight for 12 hours before and after LIPUS irradiation. Whole blood samples were obtained by tail vein sampling, and a commercially available handheld blood glucose meter was used to evaluate the glucose concentration every day. As Figure 6 shown, the blood glucose in the db / db LIPUS+MscL group was significantly lower than that in the db / db LIPUS group (P<0.05).

[0070] (IX) Real-Time Quantitative Polymerase Chain Reaction (qRT-PCR)

[0071] Trizol lysis solution was added to the pancreatic tissues of the mice, and total RNA was extracted according to the requirements of the Trizol kit. After determining its concentration with a Nano Drop2000 spectrophotometer, total RNA from the pancreatic tissues was extracted, and the RNA was reverse transcribed into cDNA using a reverse transcription kit. The PCR reaction solution was prepared, the template and primers were added, and PCR amplification was performed using a real-time quantitative polymerase chain reaction (qRT-PCR) instrument. The reaction conditions were: pre-denaturation at 95°C for 5 minutes, denaturation at 95°C for 5 seconds, annealing and extension at 60°C for 30 seconds, and repeated for 40 cycles. Using GAPDH as the internal reference gene, the relative expression level of the gene was calculated using the 2-ΔΔCT formula. The primer sequences are shown in Table 1. As Figure 7 shown, the q-PCR verification results of the pancreatic tissues indicated the successful expression of the MscL channel.

[0072] Table 1. qRT-PCR primer sequences

[0073]

[0074] (X) Hematoxylin-eosin (H&E) staining

[0075] The paraffin sections of the pancreas of mice in each group were dewaxed, stained with hematoxylin for 10 min, and then rinsed with tap water for 1 min; subsequently, rapid differentiation with 0.5% hydrochloric acid alcohol was performed for no more than 1 min, followed by rinsing with tap water; then dehydration with 95% alcohol was carried out for 1 min, and eosin staining was performed for 1 min; then the sections were successively immersed in 80% alcohol, 90% alcohol, 100% alcohol, xylene I solution, and xylene II solution for 5 min, sealed with transparent neutral gum, and finally, histological morphology was observed under a microscope to analyze its pathological changes.

[0076] The results were as Figure 8 shown. The islet area of mice in the db / db control group was significantly higher than that in the db / db LIPUS group. This result indicates that the islet area of mice in the db / db control group showed obvious hyperplasia, and the boundaries were irregular. Compared with mice in the db / db LIPUS group, the number of islets in mice in the db / db LIPUS+MscL group increased, and the hyperplasia of islets decreased significantly. This result indicates that LIPUS irradiation combined with the MscL channel further reduces islet hyperplasia.

[0077] (XI) Oil Red O staining

[0078] The frozen sections of the liver tissue to be tested were taken out and warmed at room temperature for 10 min. During this period, saturated Oil Red O stock solution and distilled water were mixed in a ratio of 3:2, and after standing for 10 min, the Oil Red O staining solution was obtained by filtration. The sections were fixed with formaldehyde-calcium for 10 min, then thoroughly washed with distilled water, and immersed in 60% isopropanol. Then, they were stained with the Oil Red O staining solution for 10 min, and again differentiated with 60% isopropanol until the stroma was clearly visible. After that, the excess staining solution was rinsed off with distilled water, and Mayer hematoxylin was used for counterstaining for 3 min, and washed again. Finally, the sections were sealed with neutral gum and air-dried naturally. Finally, histological morphology was observed under a microscope to analyze its pathological changes.

[0079] As Figure 9The results of Oil Red O staining showed that hepatocytes in the db / db control group were filled with fat vacuoles of different sizes and red or orange-red lipid droplets, and the cell nuclei were squeezed to the edge. Although there were still fat vacuoles in the db / db LIPUS group, the number of lipid droplets was significantly reduced, indicating that LIPUS treatment could reduce lipid accumulation in hepatocytes. Compared with the mice in the db / db LIPUS group, the liver tissue structure of the mice in the db / db LIPUS+MscL group was normal, with a small amount of lipid droplet accumulation, and the blue cell nuclei were evenly distributed. It was shown that the combination of LIPUS and optogenetic therapy had a better effect on reducing lipid accumulation in hepatocytes.

[0080] The above embodiments are only illustrative of the principles and effects of the present application, and are not used to limit the present application. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes completed by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed in the present application should still be covered by the claims of the present application.

Claims

1. A system for regulating blood glucose and / or treating diabetes based on optogenetics, characterized in that Comprising: An expression vector implantation module for implanting a vector capable of expressing a mutant of the large-conductance mechanosensitive channel into a subject for expression; A pulsed ultrasound device for outputting low-intensity pulsed ultrasound and performing low-intensity pulsed ultrasound stimulation on a target area of a subject implanted with the vector capable of expressing the mutant of the large-conductance mechanosensitive channel.

2. The system according to claim 1, wherein: The mutant of the large-conductance mechanosensitive channel is the MscL-G22S mutant.

3. The system according to claim 1 or 2, characterized in that: The vector is a viral vector rAAV / pan:EGFP-pA or a pReceiver-M56 Expression Clone inserted with a fluorescent tag protein.

4. The system according to claim 1, characterized in that: The ultrasound dose of the low-intensity pulsed ultrasound is 5 - 30 J, and the calculation formula of the ultrasound dose is: ultrasound dose = sound intensity × area × time × duty cycle; And / or, the ultrasound output frequency of the low-intensity pulsed ultrasound is 1 MHz; And / or, the duty cycle of the low-intensity pulsed ultrasound is 10% - 30%; And / or, the low-intensity pulsed ultrasound stimulation lasts for at least 10 days of irradiation, and is irradiated at least once a day, with each irradiation lasting for 10 - 30 min.

5. The system according to claim 1 or 4, characterized in that:

6. A product for regulating blood glucose and / or treating diabetes based on optogenetics, characterized in that, The target area is the pancreas and / or the liver. Comprising:

7. The product according to claim 6, wherein: A vector capable of expressing a mutant of the large-conductance mechanosensitive channel, and a pulsed ultrasound device for outputting low-intensity pulsed ultrasound.

8. The product according to claim 6, wherein: The mutant of the large-conductance mechanosensitive channel is the MscL-G22S mutant.

9. The product according to any one of claims 6 to 7, characterized in that: The vector is a viral vector rAAV / pan:EGFP-pA or a pReceiver-M56 Expression Clone inserted with a fluorescent tag protein. The operating parameters of the pulsed ultrasound device include: an ultrasound dose of 5 - 30 J, and the calculation formula of the ultrasound dose is: ultrasound dose = sound intensity × area × time × duty cycle; And / or, an ultrasound output frequency of 1 MHz; And / or, a duty cycle of 10% - 30%; And / or, a continuous irradiation time: at least 10 days, and at least once a day, with each irradiation lasting for 10 - 30 min.

10. Use of low-intensity pulsed ultrasound and a vector capable of expressing a mutant of the large-conductance mechanosensitive channel in the preparation of a product for regulating blood glucose and / or treating diabetes.

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