Use of ceramide hydrolase acer2 in the preparation of a drug for improving beta cell function, preventing and / or treating diabetes

Abstract

The application provides application of ceramide hydrolase ACER2 in preparation of a medicine for improving beta cell function, preventing and / or treating diabetes. The application proves, for the first time, through a pancreatic islet beta cell damage regeneration model and a diabetes mouse model, that ACER2 is indispensable for regulating ceramide level, improving beta cell function and diabetes pathological phenotype by specifically knocking out a metabolic enzyme ACER2 of vascular endothelial cells; and constructs extracellular vesicles overexpressing ACER2, and proves that the extracellular vesicles overexpressing ACER2 have certain application prospect in improvement of pancreatic islet beta cell function and treatment of diabetes.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of ceramide hydrolase ACER2 in the preparation of drugs for improving β-cell function and preventing and / or treating diabetes. Background Technology

[0002] Diabetes is currently the most common metabolic disease worldwide, and its incidence and prevalence continue to increase rapidly, placing enormous medical pressure on society. Diabetes is a metabolic disorder of carbohydrates, proteins, and fats caused by insufficient insulin secretion and / or impaired insulin utilization. It is a leading cause of complications such as blindness, amputation, chronic kidney disease, and cardiovascular disease. Type 2 diabetes mellitus (T2DM) accounts for nearly 90-95% of diabetes cases globally. Its main pathophysiological features are insulin resistance and initial hyperinsulinemia, followed by a gradual decline in the ability of pancreatic β-cells to produce insulin, leading to relative insulin deficiency and hyperglycemia. Sustained hyperglycemia leads to β-cell glucolipotoxicity, cell exhaustion, and dysfunction. Type 1 diabetes mellitus (T1DM) is common in children, accounting for 5-10% of cases, primarily due to autoimmune reactions that destroy pancreatic β-cells, resulting in complete or near-complete loss of insulin secretion.

[0003] There are various existing treatment options for diabetes, mainly including drug therapy, lifestyle interventions, and insulin replacement therapy. However, these options still face challenges in terms of long-term efficacy and patient adherence. For example, injecting exogenous insulin cannot improve pancreatic islet cell function; oral traditional hypoglycemic drugs cannot effectively maintain the content of endogenous β-cells, and some drugs have side effects; lifestyle interventions, mainly based on diet control, exercise management, and weight management, have limited efficacy because many patients find it difficult to adhere to them long-term; stem cell therapy has certain problems such as immune rejection and low tissue compatibility. These diabetes treatment options are insufficient to effectively control the progression of diabetes, mainly due to the inability to reverse the pathological process, poor control of complications, side effects and adherence issues, and insufficient individualized treatment. Therefore, there is an urgent need to find new targets and methods that can restore β-cells and improve β-cell function to treat diabetes.

[0004] Ceramide is an important biologically active sphingolipid messenger molecule that plays a crucial role in cell membrane stability, signal transduction, and stress response. Abnormal and dysregulated sphingolipid metabolism has been considered a significant driver of the progression of various metabolic diseases in recent years. Ceramide accumulation is closely related to insulin resistance, pancreatic β-cell apoptosis, and chronic inflammation, participating in the occurrence and development of insulin resistance and diabetic complications. The metabolic homeostasis of ceramide is maintained by both its synthesis and hydrolysis. The hydrolysis of ceramide is catalyzed by ceramidinases. Ceramidinases are classified into acidic, neutral, and alkaline ceramidinase subtypes based on their optimal pH for catalytic activity. Among them, alkaline ceramidinase 2 (ACER2), mainly expressed in pancreatic vascular endothelial cells, is an important member of the alkaline ceramidinase (ACER) family. It is primarily responsible for hydrolyzing specific types of ceramide into sphingosine and free fatty acids in an alkaline environment. Sphingosine can be further phosphorylated to sphingosine-1-phosphate (S1P). S1P is an important molecule promoting cell survival and inhibiting apoptosis. Ceramide hydrolysis is an important process in the pathological regulation of diabetes. Targeting ceramide and reducing ceramide levels can improve insulin sensitivity, reduce pancreatic β-cell damage, reduce inflammation, and reduce the risk of diabetic complications. Summary of the Invention

[0005] In view of this, the present invention aims to overcome the deficiencies in the prior art and proposes the application of ceramide hydrolase ACER2 in the preparation of drugs for improving β-cell function and preventing and / or treating diabetes.

[0006] To achieve the above objectives, the technical solution of the present invention is implemented as follows:

[0007] This invention provides the application of ceramide hydrolase ACER2 as a target in the preparation of drugs to improve β-cell function and prevent and / or treat diabetes.

[0008] In some specific embodiments of the present invention, the target is vascular endothelial cell ceramide hydrolase ACER2.

[0009] This invention provides the application of a ceramide hydrolase ACER2 overexpression reagent in the preparation of drugs for improving β-cell function and preventing and / or treating diabetes.

[0010] In some specific embodiments of the present invention, the ceramide hydrolase ACER2 overexpression reagent is used in any one of the following (A)-(G):

[0011] (A) Preparation of drugs that promote the regeneration of pancreatic β cells;

[0012] (B) To prepare drugs that reduce pancreatic β-cell damage and apoptosis;

[0013] (C) To prepare drugs that lower blood glucose levels in individuals with diabetes;

[0014] (D) To prepare drugs that enhance the blood glucose regulation capacity of individuals with diabetes;

[0015] (D) To prepare drugs that increase serum insulin levels in individuals with diabetes;

[0016] (E) To prepare drugs that increase the number of pancreatic β cells in diabetic individuals;

[0017] (F) Preparation of a drug that reduces serum ceramide levels in diabetic mice;

[0018] (G) Prepare a drug that increases the serum sphingosine-1-phosphate (S1P) level in individuals with diabetes.

[0019] In some specific embodiments of the present invention, the ceramide hydrolase ACER2 overexpression reagent includes functions of increasing ACER2 expression, increasing ACER2 secretion, overexpressing the ACER2 gene, and improving ACER2 activity.

[0020] In some specific embodiments of the present invention, the ceramide hydrolase ACER2 overexpression reagent comprises extracellular vesicles that overexpress ceramide hydrolase ACER2.

[0021] In some specific embodiments of the present invention, the ceramide hydrolase ACER2 overexpression reagent is an extracellular vesicle derived from human embryonic kidney cells that overexpress ACER2.

[0022] In some specific embodiments of the present invention, the diabetes is type 1 diabetes and / or type 2 diabetes.

[0023] Compared with existing technologies, the present invention has the following advantages:

[0024] This invention targets ACER2 in vascular endothelial cells to investigate its effects on β-cell function and diabetes treatment. For the first time, this invention utilizes a pancreatic β-cell damage and regeneration model and a diabetic mouse model to demonstrate the indispensable role of ACER2 in improving β-cell function and the pathological phenotype of diabetes by specifically knocking out the metabolic enzyme ACER2 in vascular endothelial cells. Extracellular vesicles overexpressing ACER2 were constructed, demonstrating their potential application in diabetes treatment. The metabolic enzyme ACER2 holds promise as a novel target for diabetes treatment and its clinical application. Attached Figure Description

[0025] Figure 1A schematic diagram illustrating the establishment of a β-cell injury regeneration model and the Acer2 gene knockout strategy in mouse vascular endothelial cells (using tamoxifen to induce Acer2 deletion in mouse vascular endothelial cells). WT As a control group, Acer2 i ΔEC The experimental group, namely the Acer2 knockout group of vascular endothelial cells, was used to induce pancreatic β-cell damage using streptozotocin (STZ). Cells were collected and analyzed after 2 weeks.

[0026] Figure 2 To detect the β-cell content in the control and experimental groups in a β-cell injury regeneration model (A. Detection of insulin and glucagon expression in pancreatic sections of mice in both groups by immunofluorescence staining; B. Statistical analysis of insulin and glucagon volume in mice in both groups, showing that the number of pancreatic β-cells was significantly reduced after knocking out Acer2 in vascular endothelial cells).

[0027] Figure 3 To detect the degree of cell damage and senescence in the control and experimental groups of a β-cell injury regeneration model (A. Immunofluorescence staining was used to detect the expression levels of insulin and the cell damage marker γH2A.X in pancreatic sections of mice in both groups; B. The number of damaged cells in pancreatic islets in both groups of mice was statistically analyzed, showing that knocking out Acer2 in vascular endothelial cells significantly increased the total number of damaged cells and the number of damaged β-cells in the islets; C. Immunofluorescence staining was used to detect the expression levels of insulin and the cell senescence marker p21 in pancreatic sections of mice in both groups of mice; D. The number of senescent cells in pancreatic islets in both groups of mice was statistically analyzed, showing that knocking out Acer2 in vascular endothelial cells significantly increased the total number of p21-positive senescent cells and the number of senescent β-cells in the islets).

[0028] Figure 4 To detect the degree of cell proliferation in the control and experimental groups in a β-cell injury regeneration model (A. Detection of insulin, vascular marker CD31 and cell proliferation marker EdU in pancreatic slices of mice in both groups by immunofluorescence staining; B. Statistical analysis of the number of proliferating cells in pancreatic islets of mice in both groups, showing that the number of EdU-positive proliferating β-cells in pancreatic islets was significantly reduced after knocking out Acer2 in vascular endothelial cells).

[0029] Figure 5 A schematic diagram illustrating the strategy for establishing a diabetic mouse model and knocking out Acer2 in mouse vascular endothelial cells (using tamoxifen to induce Acer2 deletion in mouse vascular endothelial cells). WT As a control group, Acer2 iΔEC The experimental group consisted of Acer2 knockout vascular endothelial cells. Diabetes was induced by five injections of streptozotocin (STZ) as indicated, and samples were collected after 12 weeks for analysis.

[0030] Figure 6 The changes in blood glucose in the control and experimental groups of diabetic mice were shown in the graph (A. Changes in blood glucose in the two groups of diabetic mice after modeling; B. Statistical graph of area under the blood glucose curve, showing that deletion of Acer2 in vascular endothelial cells significantly increased blood glucose in diabetic mice).

[0031] Figure 7 The dynamic changes in glucose tolerance in the control and experimental groups of diabetic mice were shown in the following figures: (A. Dynamic monitoring of blood glucose levels in the two groups of mice after intraperitoneal injection of glucose; B. Statistical plot of the area under the curve of blood glucose changes after glucose injection in the two groups of mice, showing that the glucose tolerance of diabetic mice decreased after deletion of Acer2 in vascular endothelial cells; C. Serum insulin levels in the two groups of mice after glucose injection).

[0032] Figure 8 To detect the β-cell content in the control and experimental groups of a diabetic mouse model (A. Detection of insulin and glucagon expression in pancreatic sections of the two groups of mice by immunofluorescence staining; B. Statistical analysis of insulin and glucagon volume in the two groups of mice, showing that the number of pancreatic β-cells in diabetic mice was significantly reduced after knocking out Acer2 in vascular endothelial cells).

[0033] Figure 9 To detect the levels of ceramide and its hydrolysis products in the serum of control and experimental mice in a diabetic mouse model (A. Comparison of ceramide levels in the serum of the two groups of mice showed that the ceramide level in the serum of diabetic mice was significantly increased after Acer2 knockout in vascular endothelial cells; B. Comparison of dihydroceramide levels in the serum of the two groups of mice showed that the dihydroceramide level in the serum of diabetic mice was significantly increased after Acer2 knockout in vascular endothelial cells; C. Comparison of sphingosine-1-phosphate (S1P) levels in the serum of the two groups of mice showed that the sphingosine-1-phosphate (S1P) level in the serum of diabetic mice was significantly increased after Acer2 knockout in vascular endothelial cells).

[0034] Figure 10 In a diabetic mouse model, the control group injected with physiological saline (Saline) and the treatment group treated with extracellular vesicles derived from human embryonic kidney cells (HEK293) (EV) were compared. 293 The treatment group consisted of extracellular vesicles derived from human embryonic kidney cells (HEK 293T) that overexpressed ACER2 (EV). 293AC Blood glucose changes in mice (A. Blood glucose changes in three groups of diabetic mice after modeling and treatment; B. Area under the blood glucose curve, showing EV) 293AC The blood glucose levels of diabetic mice in the treatment group were significantly reduced.

[0035] Figure 11In a diabetic mouse model, the control group injected with physiological saline (Saline) and the treatment group treated with extracellular vesicles derived from human embryonic kidney cells (HEK293) (EV) were compared. 293 The treatment group consisted of extracellular vesicles derived from human embryonic kidney cells (HEK 293T) that overexpressed ACER2 (EV). 293AC Dynamic changes in glucose tolerance in mice (A. Changes in blood glucose levels at different time points in three groups of mice after intraperitoneal injection of glucose; B. Statistical graph of area under the blood glucose curve, showing EV) 293AC The treatment group mice showed improved glucose tolerance.

[0036] Figure 12 In a diabetic mouse model, the control group injected with physiological saline (Saline) and the treatment group with extracellular vesicles derived from human embryonic kidney cells (HEK293T) (EV) were compared. 293 ) and the extracellular vesicle therapy group derived from human embryonic kidney cells (HEK293) that overexpress ACER2 (EV) 293AC Detection of pancreatic β-cell content in mice (A. Detection of insulin and glucagon expression in pancreatic sections of three groups of mice by immunofluorescence staining; B. Statistical analysis of insulin content in the three groups of mice, showing EV...) 293AC The number of pancreatic β cells was significantly increased in the treatment group of diabetic mice. Detailed Implementation

[0037] Unless otherwise defined, the technical terms used in the following embodiments have the same meanings as commonly understood by those skilled in the art to which this invention pertains. Unless otherwise specified, the experimental reagents used in the following embodiments are conventional biochemical reagents; and the experimental methods described are conventional methods.

[0038] Twelve-week-old SPF-grade male wild-type C57BL / 6J mice were obtained from Vital River Pharmaceuticals, Beijing. (Acer2) fl / fl The mice were from Shanghai Nanmo Biotechnology Co., Ltd., and the Cdh5-(PAC)-CreERT2 mice were from Jackson Biotechnology Co., Ltd. in the United States.

[0039] All mice were housed in the SPF-grade animal facility at Tianjin Medical University, with an indoor temperature of 25°C, 12-hour continuous light, and ample food and water. All research protocols involving animal use were approved by the Institutional Animal Care and Use Committee of Tianjin Medical University. All experimental procedures complied with the National Institutes of Health's Laboratory Animal Care and Use Guidelines.

[0040] Anti-Insulin antibody (ab181547) (Abcam); Anti-Glucagon antibody (MAB1249) (B&D); Anti-γH2A.X antibody (9718S) (CST); Anti-p21 antibody (Ab188224) (Abcam); Surgical microscope (Motic); Microinjector (Hamilton, USA); LSM900 laser confocal microscope (Zeiss); Insulin assay kit (EZ assay).

[0041] The specific embodiments provided by the present invention will be described in detail below with reference to the accompanying drawings.

[0042] 1. A method for specific knockout of Acer2 in mouse vascular endothelial cells:

[0043] Mice using the Cre-LoxP recombinase system will have Acer2 fl / fl Mice were bred with Cdh5-(PAC)-CreERT2 mice to obtain Acer2. fl / fl Cdh5-(PAC)-CreERT2 mice were induced to have Acer2 knockout in vascular endothelial cells by intraperitoneal injection of tamoxifen at 7, 5, 3, and 1 days before modeling, respectively. iΔEC Mice (e.g.) Figure 1 (As shown).

[0044] 2. Drug-induced β-cell injury and regeneration model method:

[0045] Six-week-old male C57BL / 6J mice were weighed and then injected intraperitoneally with streptozotocin (STZ) at a dose of 100 mg / kg. Samples were collected two weeks later for various tests.

[0046] 3. Immunofluorescence staining of pancreatic sections

[0047] (1) Experimental Methods: Mice were euthanized by CO2 anesthesia, and pancreatic samples were collected. After fixation with 4% paraformaldehyde for 1 hour, the samples were embedded in OCT and sectioned to a thickness of 10 μm. The sections were permeabilized with PBS containing 0.3% Triton for 20 minutes, and then incubated in blocking buffer (PBS containing 0.3% Triton and 2% BSA) for 1 hour. Subsequently, the samples were incubated overnight at 4°C with the first antibody (diluted in blocking buffer, 1:200). The next day, the samples were washed three times with PBS and incubated at room temperature for 2 hours with an appropriate concentration of secondary antibody (diluted in blocking buffer, 1:300). After washing three times with PBS, the samples were mounted with neutral resin. Images were taken using a Zeiss LSM 900 confocal microscope. Quantitative analysis was performed using ImageJ software.

[0048] (2) Experimental results:

[0049] like Figure 2 Insulin and glucagon staining results of the pancreas in the β-cell injury regeneration model mice shown in A and 2B indicate that knocking out Acer2 in the vascular endothelial cells of the experimental group significantly reduced the number of β-cells. Figure 3 The staining results of pancreatic insulin and the cell damage marker γH2A.X in the β-cell damage regeneration model mice shown in A and 3B indicate that the total number of damaged cells and the number of damaged β-cells were significantly reduced in the experimental group, suggesting that knocking out Acer2 in vascular endothelial cells significantly aggravated pancreatic β-cell damage; Figure 3 The staining results of pancreatic insulin and the cellular senescence marker p21 in the β-cell injury regeneration model mice shown in C and 3D indicate that knocking out Acer2 in the vascular endothelial cells of the experimental group significantly aggravated cellular senescence; as shown in Figures C and 3D. Figure 4 The results of pancreatic insulin, vascular marker CD31, and cell proliferation marker EdU staining in the β-cell injury regeneration model mice shown in A and 4B indicate that the number of EdU-positive proliferating β-cells in the islets of Langerhans was significantly reduced after Acer2 knockout in the vascular endothelial cells of the experimental group; Figure 8 The insulin and glucagon staining results of the pancreas in the diabetic mouse models shown in A and 8B indicate that knocking out Acer2 in vascular endothelial cells in the experimental group reduced the number of pancreatic β cells in diabetic mice; Figure 12 The results of insulin and glucagon staining of the pancreas in diabetic mouse models shown in A and 12B indicate EV 293AC The number of pancreatic β cells was significantly increased in the treatment group of diabetic mice.

[0050] 4. Methods for establishing a drug-induced diabetic mouse model:

[0051] Twelve-week-old male C57BL / 6J mice were weighed and then intraperitoneally injected with streptozotocin (STZ) at a dose of 50 mg / kg for 5 consecutive days. Blood glucose levels were measured two weeks later. Mice with a non-fasting blood glucose level ≥16.8 mM were considered to have successfully established the mouse model (e.g., Figure 5 (As shown).

[0052] 5. Blood glucose monitoring and glucose tolerance test:

[0053] (1) Detection method: Acer2 after successful diabetes modeling WT and Acer2 iΔEC Mice underwent weekly tail vein blood glucose measurements for 8 weeks, followed by a glucose tolerance test. After successful establishment of a diabetic model, saline and EV were administered. 293 and EV 293AC In the treated mice, blood glucose levels were measured twice weekly via tail vein sampling for 9 weeks, followed by a glucose tolerance test. In the glucose tolerance test, mice were starved for 8 hours and then injected intraperitoneally with a 10% glucose solution. Subsequently, blood glucose levels were measured via tail vein sampling at 0, 15, 30, 60, and 120 minutes, and statistical analysis was performed. Serum was then collected from the collected venous blood to determine the insulin content.

[0054] The culture method for human embryonic kidney cells (HEK 293T) was as follows: Human embryonic kidney cells (HEK 293T) were used in this example and cultured in DMEM medium containing 10% bovine serum, 100 U / mL penicillin, and 100 mg / mL streptomycin. Subsequently, a plasmid overexpressing the ACER2 gene was transfected into HEK 293T cells using a transfection reagent to overexpress the ACER2 protein.

[0055] The method for extracting extracellular vesicles is as follows: Collect cell supernatant, centrifuge at 1000g for 10 minutes, take the supernatant and centrifuge at 10,000g for 30 minutes, filter the supernatant after centrifugation using a 0.22μm filter, and then centrifuge at 100,000g for 1 hour using an ultracentrifuge. After centrifugation, discard the supernatant, resuspend in a large volume of PBS, centrifuge again at 100,000g for 90 minutes, discard the supernatant, and resuspend in an appropriate volume of PBS buffer for later use.

[0056] (2) Experimental results:

[0057] like Figure 6 The blood glucose changes and area under the blood glucose curve (AUC) plots shown in Figures A and 6B for the control and experimental groups of diabetic mice demonstrate that deletion of Acer2 in vascular endothelial cells significantly exacerbated hyperglycemia in diabetic mice. Figure 7The dynamic changes in glucose tolerance, statistical results of the area under the blood glucose curve, and the detection results of serum insulin levels shown in A-7C indicate that the glucose tolerance of diabetic mice after deletion of Acer2 in vascular endothelial cells is reduced, and the serum insulin level of the mice is also significantly reduced; Figure 10 The graphs A and 10B show the blood glucose changes and area under the blood glucose curve in diabetic mouse models under three treatments, illustrating EV (extracorporeal membrane oxygenation). 293AC The hyperglycemia in the treated diabetic mice was significantly improved; such as Figure 11 The dynamic changes in glucose tolerance and the statistical results of the area under the blood glucose curve shown in A and 11B indicate that EV 293AC The glucose tolerance of diabetic mice in the treatment group was significantly enhanced.

[0058] 6. Analyze serum ceramide and sphingosine-1-phosphate (S1P) levels using metabolomics:

[0059] (1) Detection Method: After blood glucose monitoring and glucose tolerance tests were completed in diabetic mice, serum samples from each group of mice were collected for targeted lipidomics analysis using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS). 50 μL of serum was mixed with 400 μL of pre-chilled 75% HPLC-grade methanol containing 17:0 ceramide (0.2 μL, 1 μg / μL), S1P d7 (1 μL, 1 μg / μL), or sphingosine d7 (0.1 μL, 0.1 μg / μL) and 1 mL of MTBE to extract the sample. The sample was vortexed for 20 minutes and then allowed to stand at room temperature for 10 minutes. Subsequently, it was centrifuged at 12,000 rpm for 10 minutes at 4 °C, and the upper lipid phase was transferred and dried under nitrogen. Then, 150 μL of methanol was added, and the mixture was vortexed at room temperature for 10 minutes. The supernatant was transferred to a filtered centrifuge tube (Corning, 8169) and centrifuged at 12,000 g for 10 minutes at 4 °C. Subsequently, 2 μL of the filtered sample was used for the detection of ceramide, dihydroceramide, and S1P, and injected into an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS / MS) system for analysis. The raw data generated by the UHPLC-MS system were analyzed using Analyst 1.6 software, and analyte quantification was performed using MultiQuant 3.0.2 software. Multivariate statistical analysis and data visualization were performed using MetaboAnalyst 6.0, including t-tests, cluster analysis, partial least squares discriminant analysis (PLS-DA), and heatmaps. Quantitative results are presented as mean ± standard error (SEM), and graphs were generated using GraphPad Prism.

[0060] (2) Experimental results:

[0061] like Figure 9 Targeted lipidomics analysis of serum samples from the control and experimental groups of diabetic mouse models shown in A-9C yielded a comparison of the levels of ceramide, dihydroceramide, and sphingosine-1-phosphate. The results showed that deletion of Acer2 cells in vascular endothelial cells significantly increased the levels of ceramide and dihydroceramide in the serum of diabetic mice and significantly reduced the content of sphingosine-1-phosphate.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Application of ceramide hydrolase ACER2 as a target in the preparation of drugs to improve β-cell function and prevent and / or treat diabetes.

2. The application according to claim 1, characterized in that: The target is vascular endothelial cell ceramide hydrolase ACER2.

3. Application of ceramide hydrolase ACER2 overexpression reagent in the preparation of drugs to improve β-cell function and prevent and / or treat diabetes.

4. The application according to claim 3, characterized in that: The use of the ceramide hydrolase ACER2 overexpression reagent in any one of the following (A)-(G): (A) Preparation of drugs that promote the regeneration of pancreatic β cells; (B) To prepare drugs that reduce pancreatic β-cell damage and apoptosis; (C) To prepare drugs that lower blood glucose levels in individuals with diabetes; (D) To prepare drugs that enhance the blood glucose regulation capacity of individuals with diabetes; (D) To prepare drugs that increase serum insulin levels in individuals with diabetes; (E) To prepare drugs that increase the number of pancreatic β cells in diabetic individuals; (F) Preparation of a drug to reduce serum ceramide levels in diabetic mice; (G) Prepare drugs that increase serum sphingosine-1-phosphate levels in individuals with diabetes.

5. The application according to claim 3, characterized in that: The ceramide hydrolase ACER2 overexpression reagent has the functions of increasing ACER2 expression, increasing ACER2 secretion, overexpressing the ACER2 gene, and improving ACER2 activity.

6. The application according to claim 3, characterized in that: The ceramide hydrolase ACER2 overexpression reagent comprises extracellular vesicles that overexpress ceramide hydrolase ACER2.

7. The application according to claim 3, characterized in that: The diabetes mellitus refers to type 1 diabetes and / or type 2 diabetes.

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