Application of fer1l6 gene in prevention and treatment of diabetes

Abstract

This invention belongs to the fields of biomedicine and molecular biology, specifically relating to the application of the FER1L6 gene in the prevention and treatment of diabetes. In this application, the expression of GLUT4 is increased by inhibiting the expression of FER1L6, thereby improving the related physiological metabolism in diabetic organisms. This physiological metabolism includes glucose uptake, lipid metabolism, and oxidative stress damage. Experimental results show that FER1L6 is highly expressed in type 2 diabetic mice and insulin-resistant HepG2 cells; reducing FER1L6 expression significantly increases GLUT4 expression, ultimately improving lipid metabolism and oxidative stress damage in diabetic mice, while also enhancing cellular glucose uptake. Therefore, it can be concluded that FER1L6 is a good regulatory target for the prevention and treatment of type 2 diabetes based on the GLUT4 pathway. Based on this target, new technical approaches can be provided for the prevention and control of type 2 diabetes.

Description

Technical Field

[0001] This invention belongs to the fields of biomedicine and molecular biology, specifically relating to the application of the FER1L6 gene in the prevention and treatment of diabetes. Background Technology

[0002] Diabetes mellitus is a metabolic disease characterized by hyperglycemia. Clinically, based on clinical symptoms, age, and other factors, it is mainly divided into two categories: type 1 diabetes and type 2 diabetes (T2DM). Type 2 diabetes (T2DM) is characterized by chronic hyperglycemia.

[0003] The direct cause of diabetes is primarily insufficient insulin secretion or decreased peripheral insulin sensitivity. With a large affected population, it has become a significant global public health concern. The common treatment strategy for diabetes involves a combination of diet, exercise, and medication. Clinically, medication primarily involves oral hypoglycemic agents and insulin injections. However, hypoglycemia is a common side effect of these drugs, and long-term insulin injections may increase insulin resistance. Therefore, early prevention remains the main strategy for diabetes management, and developing new therapeutic targets is a major technological focus in diabetes prevention and control.

[0004] Insulin resistance is a common issue in the prevention and treatment of type 2 diabetes mellitus (T2DM). Enhancing insulin sensitivity is a common and effective strategy for addressing this condition. Studies of related metabolic pathways have shown that glucose uptake after insulin stimulation requires the mediation of glucose transporter 4 (GLUT4). GLUT4 possesses an extremely long half-life (over 24 hours) and a slow reuptake rate, characteristics that contribute to its stability and targeting during insulin stimulation. Studies in T2DM patients have revealed that one of the hallmarks of T2DM is impaired GLUT4 membrane translocation and reduced expression levels. Related mouse experiments have shown that GLUT4 knockout mice exhibit symptoms of insulin resistance and glucose intolerance, displaying a diabetic phenotype; while mice overexpressing GLUT4 show improved insulin action and reduced blood glucose levels in both fasting and postprandial states. Based on these findings, further research into GLUT4 expression-related proteins can provide more technical insights for the prevention and treatment of T2DM. Summary of the Invention

[0005] Based on existing research on the Ferlin protein family, using an insulin-resistant cell model as experimental material, and considering the importance of GLUT4 in glucose transport, the inventors believe that FER1L6 is an important target for the prevention and treatment of type 2 diabetes. Based on this result, a good technical foundation can be laid for further improvement of the relevant insulin metabolism regulation mechanism and the development of new diabetes drugs.

[0006] The technical solution adopted in this application is described in detail below.

[0007] Application of FER1L6 gene in the prevention and treatment of diabetes, wherein diabetes is type 2 diabetes (T2DM); FER1L6 is highly expressed in type 2 diabetic organisms and insulin-resistant HepG2 cells. Inhibiting the expression of FER1L6 can significantly increase the expression of GLUT4, thereby improving lipid metabolism and oxidative stress damage in diabetic organisms.

[0008] The lipid metabolism-related genes include, but are not limited to, triglycerides (TG) and total cholesterol (TC).

[0009] The oxidative stress indicators include, but are not limited to, superoxide dismutase (SOD) and malondialdehyde (MDA).

[0010] The specific methods for inhibiting FER1L6 expression include: designing RNA interference molecules or antisense oligonucleotides, small molecule inhibitors, or siRNAs targeting FER1L6, and inhibiting FER1L6 transcription and expression by implementing lentiviral infection or gene knockout; or designing specific antibodies against FER1L6 itself or its upstream and downstream molecules (i.e., anti-FER1L6 antibodies).

[0011] Previous studies have shown that the Ferlin protein family includes at least six members: Dysferlin (FER1L1), Otoferlin (FER1L2), Myoferlin (FER1L3), FER1L4, FER1L5, and FER1L6, which is the subject of this application. These proteins share common structural features, including multiple Ca²⁺ ions. 2+ Combining the C2 domain, the FerA domain, and a transmembrane domain that is membrane-anchored via its single C-terminus, this family of proteins is believed to function in Ca2+. 2+ These proteins play crucial roles in triggered membrane fusion and vesicle transport, thus this gene family is closely associated with the development of various diseases. Studies on the association between this protein family and insulin have shown that FER1L3 mediates the endocytic cycle and participates in the transport of insulin-like growth factor receptors; Fer1L5 directly binds to the endocytic cycle proteins EHD1 and EHD2, and EHD2 may be involved in the insulin-induced transport of GLUT4 to the cell membrane in rat adipocytes. However, regarding the function of FER1L6, only some studies have reported on its role in muscle formation during zebrafish development; no studies have yet reported on whether FER1L6 is related to glucose metabolism or even the prevention and treatment of type 2 diabetes mellitus (T2DM).

[0012] Based on existing research on the Ferlin protein family, the inventors conducted experimental studies on the application prospects of FER1L6 in glucose metabolism and type 2 diabetes mellitus (T2DM). Preliminary experimental results showed that FER1L6 was highly expressed in type 2 diabetic mice and insulin-resistant HepG2 cells (which preliminarily suggests that elevated FER1L6 levels lead to or accompany the development of diabetes). Further in vivo experiments by knocking down FER1L6 showed that reducing FER1L6 expression significantly increased GLUT4 expression, ultimately improving lipid metabolism and oxidative stress damage in diabetic mice. Related cell experiments also confirmed that reducing FER1L6 expression can improve cellular glucose uptake.

[0013] Based on the relevant experimental evidence in this application, the inventors believe that FER1L6 is a good regulatory target for the prevention and treatment of type 2 diabetes based on the GLUT4 pathway. This target can provide new technical approaches for the prevention and control of type 2 diabetes, and also offers valuable insights and references for the development of other types of drugs. Attached Figure Description

[0014] Figure 1 These are the experimental results related to the construction of the insulin resistance cell model; among which:

[0015] A shows the differences in glucose uptake in HepG2 cells after treatment with different concentrations of GlcN;

[0016] B represents the validation results of insulin treatment on the insulin-resistant cell model;

[0017] C is a differential gene volcano plot after transcriptomic analysis of GlcN-vs-Control cells;

[0018] D represents further detection of the expression levels of the top 5 genes with significant differences in expression using qRT-PCR; Note: Compared with the control group, * P <0.05,** P <0.01, *** P <0.001; compared with the LPS group, # P <0.05, ## P <0.01, ### P <0.001;

[0019] Figure 2 These are the experimental results related to gene silencing FER1L6; among them:

[0020] A represents the result of FER1L6 expression level detection after gene silencing;

[0021] B represents the results of glucose uptake detection after gene silencing;

[0022] C represents the result of detecting GLUT4 gene expression after gene silencing;

[0023] D represents the result of GLUT4 protein expression detection after gene silencing; Note: Compared with the control group, * P <0.05,** P <0.01, *** P <0.001; compared with the model group, # P <0.05, ## P <0.01, ### P <0.001;

[0024] Figure 3 The results of animal experiments after knocking down FER1L6 in mouse livers; among which:

[0025] A represents the expression levels of FER1L6 and GLUT4 detected using Western blot technology.

[0026] B represents the protein quantification result of FER1L6;

[0027] C represents the protein quantification result of GLUT4;

[0028] D represents the HE and Oil Red O staining results of mouse liver;

[0029] E represents the results of serum triglyceride and total cholesterol levels in mice;

[0030] F represents the levels of superoxide dismutase and malondialdehyde in mouse liver; Note: Compared with the control group, * P <0.05,** P <0.01, *** P <0.001; compared with the model group, # P <0.05, ## P <0.01, ### P <0.001;

[0031] In the figure: "HFD" represents the high-fat diet group; AAV8-shCtrl represents the gene silencing control group; Alloxan represents the model group; and AAV8-shFeril6 represents the gene silencing group. Detailed Implementation

[0032] The present application will be further explained below with reference to the embodiments. Before introducing the specific embodiments, the experimental background of some embodiments is briefly described below.

[0033] Biomaterials:

[0034] HepG2 cells (liver cancer cells) were purchased from the Cell Bank of the Chinese Academy of Sciences Type Culture Collection Committee. The culture conditions were as follows: cultured in DMEM medium containing a mixture of 10% fetal bovine serum and 1% penicillin and streptomycin at 37°C in a 5% CO2 incubator.

[0035] The synthesis of the relevant primers and the sequencing work were completed by BGI Genomics Co., Ltd.

[0036] C57BL / 6 male mice (4 weeks old) were purchased from Liaoning Changsheng Biotechnology Co., Ltd. (SCXK(Liaoning)2020-0001).

[0037] Example 1

[0038] Given the significant technological importance of diabetes prevention and treatment, the inventors constructed a cell model of insulin resistance and screened and identified key regulatory genes to lay a technological foundation for diabetes prevention and treatment or the development of novel drugs. A brief overview of the relevant experiments is as follows.

[0039] (I) Construction of an insulin-resistant cell model

[0040] HepG2 cells were loaded at a rate of 4 × 10 4 The cells were seeded at a density of 1 cell per well in 96-well plates and cultured for about 24 hours until they adhered. Then, the complete culture medium was replaced with serum-free high-glucose DMEM medium and cultured for another 12 hours (to starve the cells).

[0041] Subsequently, the cells were divided into different treatment groups:

[0042] Blank control group: Discard the old culture medium and add new DMEM culture medium;

[0043] Experimental group (insulin resistance model group): The culture medium was replaced with DMEM medium containing different concentrations of GlcN (glucosamine), with the concentrations of GlcN set as 1, 4, 8, 12, 16 and 20 mmol / L, respectively.

[0044] After the relevant treatment groups were placed in an incubator for 24 h of further culture, the glucose content in the cell supernatant was detected by the GOD-POD method, and the amount of glucose taken up by the cells during this period was calculated.

[0045] Experimental results show (results as follows) Figure 1 of Figure 1A): Treatment of HepG2 cells with different concentrations of GlcN significantly inhibited the cells' consumption of glucose. P <0.001), and this inhibitory effect is dose-dependent.

[0046] Based on the aforementioned GlcN treatment, the cells in different experimental groups were replaced with DMEM medium supplemented with insulin (referencing existing technical literature and considering factors such as ease of operation, the insulin dosage was set to 3 µM). After culturing for another 24 h, the amount of glucose taken up by the cells was detected and calculated (to assess whether the insulin resistance model based on HepG2 cells was successful).

[0047] The results showed that ( Figure 1 B): Compared with the blank control group, the addition of insulin significantly promoted glucose uptake by HepG2 cells. P <0.001); Compared with the control group treated with insulin, the 8, 12, and 16 mM GlcN treatment groups still failed to significantly promote cellular glucose uptake after insulin treatment. This indicates that GlcN treatment induced insulin resistance in the cells, and even with insulin, the cellular glucose uptake and utilization rate could not be improved. Based on this result, subsequent experiments used 8 mM GlcN treatment for 24 h to induce insulin resistance in HepG2 cells.

[0048] (ii) Further sequencing analysis of the constructed insulin resistance model

[0049] To explore novel therapeutic targets for diabetes, further transcriptomic mRNA quantification was performed on the aforementioned HepG2 and IR-HepG2 cells (insulin-resistant cells). Based on the results, differentially expressed genes were screened using a corrected Q < 0.001. The results are as follows: Figure 1 As shown in C.

[0050] Analysis results showed that 68 differentially expressed genes were upregulated and 199 differentially expressed genes were downregulated in the GlcN-vs-Control differential results. This result also means that these differentially expressed genes may be potential target genes involved in insulin resistance in HepG2 cells (as for the FER1L6 gene, FER1L6 is highly expressed in insulin-resistant HepG2 cells, ranking third in differential expression, which indicates that elevated FER1L6 levels may lead to or accompany the development of diabetes).

[0051] To further narrow down the target gene pool, the inventors selected the top 5 genes that were significantly upregulated and used qRT-PCR technology to detect the differences in expression levels of these 5 genes between insulin-resistant and control cells. The results showed (e.g.) Figure 1 As shown in D): FER1L6, CYP3A7-CYP3A5IP, KNG1, HPX, and UGT2B15 were all significantly upregulated in the insulin-resistant cell model group. P The difference was <0.001), and FER1L6 had the highest difference fold (about 30 times). For this reason, the inventors chose FER1L6 as the object of further in-depth research.

[0052] Example 2

[0053] Based on the identification of the FER1L6 gene as the research object in Example 1, and the preliminary clarification that high expression of FER1L6 has a direct impact on glucose uptake, and combined with existing research on the FER1L6 gene, the inventors further utilized siRNA treatment technology to reduce the expression level of FER1L6 in IR-HepG2 cells to further verify the function of this gene. A brief description of the relevant experimental process is as follows.

[0054] (I) Construction of siRNA primers

[0055] Based on siRNA technology, the following primer sequences were designed targeting FER1L6-Homo-1680:

[0056] Positive: 5'- CCGGAAGAUUGGAGAUAAATT-3',

[0057] Reverse: 5'- UUUAUCUCCAAUCUUCCGGTT-3';

[0058] As a negative control, the relevant primer sequences were designed as follows:

[0059] Positive: 5'- UUCUCCGAACGUGUCACGUTT-3',

[0060] Reverse: 5'- ACGUGACACGUUCGGAGAATT-3';

[0061] (ii) siRNA transfection

[0062] Using Lipo2000 as the transfection reagent, siRNA primers (provided by BGI Genomics in lyophilized powder form, ready to use after dissolution) were introduced into IR-HepG2 cells. For specific procedures, refer to the transfection reagent instructions, or refer to the following:

[0063] Pre-prepared IR-HepG2 cells at a ratio of 1×10 5 Inoculate at a density of cells / well into 24-well plates and incubate for 24 h in preparation for transfection;

[0064] During transfection, 50 μL of serum-free and antibiotic-free MEM medium and siRNA were added to an EP tube (to explore the appropriate amount of siRNA, the siRNA concentration was 20 μmol / L, and the amounts were 1, 1.5, and 2 μL, respectively), and incubated for 5 min.

[0065] Meanwhile, take another EP tube, add 50 μL of serum-free and antibiotic-free MEM medium and 1 μL of Lipo2000, and incubate for 5 min;

[0066] After mixing the solutions from the two EP tubes, incubate for 20 min to promote the formation of siRNA-cationic liposome complex.

[0067] Subsequently, discard the culture medium of HepG2 cells in the 24-well plate that was cultured in advance, add 400 μL of serum-free and antibiotic-free culture medium to each well, and 100 μL of the above-mentioned siRNA-cationic liposome complex, and gently shake to mix.

[0068] Then, continue incubation in the incubator for 6 hours (for transfection), discard the culture medium, add 500 μL of fresh serum-free and antibiotic-free culture medium to each well, and continue culturing.

[0069] Finally, the changes in glucose levels in the cell supernatant were detected at 24, 48, and 72 h of culture. Cells from the treatment group with significant changes in glucose levels were collected, and their genome (total RNA) was extracted and reverse transcribed into cDNA. The corresponding mRNA expression levels were then detected using qRT-PCR technology (with β-actin as an internal reference).

[0070] The reference primer sequence design for qRT-PCR detection is as follows:

[0071] Primers for the β-actin gene were designed as follows:

[0072] F primer: 5'-GGCTGTATTCCCCTCCATCG-3',

[0073] R primer: 5'-CCAGTTGGTAACAATGCCATGT-3';

[0074] Primers for the FER1L6 gene were designed as follows:

[0075] F primer: 5'-CCAGAGGAGTCACCAAGAAGA-3',

[0076] R primer: 5'-GGTGAGGACCAATCCCTTCTC-3';

[0077] Primers for the GLUT4 gene were designed as follows:

[0078] F primer: 5'-GTGACTGGAACACTGGTCCTA-3',

[0079] R primer: 5'-CCAGCCACGTTGCATTGTAG-3';

[0080] The reference design for a 20μL amplification system is as follows:

[0081] 2 x SYBR Green qPCR Master Mix (Low ROX), 5 μL;

[0082] F primer, 0.8 μL (10 μM);

[0083] R primer, 0.8 μL (10 μM);

[0084] Template cDNA, 10 μL;

[0085] Nuclease-Free Water, add to a final volume of 20 μL;

[0086] The reaction procedure was as follows: pre-denaturation, 95℃, 30s; 95℃, denaturation for 15s, 60℃, extension for 30s, 40 cycles; finally, the dissolution curve was obtained and the expression level was calculated.

[0087] During the experiment, two control groups were designed: a negative control (siRNA negative control) and a negative control + GLcN treatment control group (8 mM).

[0088] Experimental results show that ( Figure 2 of Figure 2 A) Under different siRNA treatment conditions (siRNA usage of 40, 60, and 80 pmol, respectively), the expression of FER1L6 gene mRNA after silencing was significantly reduced. P <0.01); however, further glucose uptake tests showed that glucose uptake varied under different siRNA treatment conditions, with a significant decrease in glucose levels only observed under specific siRNA treatment conditions (siRNA dosage of 60 pmol). Figure 2 B, P <0.05), meaning that at this dosage, the inhibitory effect on FER1L6 expression is most significant.

[0089] Further analysis using qRT-PCR and Western Blot techniques revealed the expression of the GLUT4 gene and protein after FER1L6 gene silencing (60 pmol siRNA treatment group). The results showed that... Figure 2 C Figure 2 D): In the gene silencing group, both the gene expression level and the GLUT4 protein content were significantly increased. P <0.001).

[0090] These results all indicate that knocking down the FER1L6 gene significantly increased GLUT4 expression, thereby improving glucose uptake (or improving glucose homeostasis) in IR-HepG2 cells.

[0091] Example 3

[0092] Based on the verification results of the aforementioned embodiments, the inventors further conducted relevant animal experiments by constructing a mouse model that targets and knocks down FER1L6 in the liver. The relevant experimental details are briefly described below.

[0093] Experimental procedure:

[0094] After one week of acclimatization, male C57BL / 6 mice were divided into experimental groups:

[0095] Normal group: 8 mice were given normal food and water;

[0096] Experimental group: Sixteen mice were fed a high-fat diet for four weeks and then injected intraperitoneally with alloxan (50 mg / kg) every two days for three consecutive times to establish a type 2 diabetes mellitus (T2DM) mouse model.

[0097] One week later, when the fasting blood glucose level of the mice remained above 11.1 mmol / L and the mice exhibited polydipsia, polyphagia, polyuria, and weight loss, the diabetes model was considered to have been successfully established. (One of the main differences between type 1 and type 2 diabetes is that type 1 diabetes is generally considered to be a loss of insulin secretion ability, while type 2 diabetes is characterized by insulin resistance. Therefore, the model established in this example is a type 2 diabetes model.)

[0098] Eight mice from a diabetic model were randomly selected and injected via tail vein with AAV8-GP-1-SJNC (control virus) as a control.

[0099] The other 8 animals were injected with AAV8-GP-1-FER1L6 virus particles as the FER1L6 knockdown test group;

[0100] After the injection, continue feeding with a high-fat diet for 2 months.

[0101] It should be explained that the AAV8-GP-1-FER1L6 viral plasmid (target sequence as shown in SEQ ID No. 1, specifically: CCGGAAGATTGGAGATAAA) was constructed by Gemma Corporation (the control viral plasmid was also provided by the company). Before application, the viral plasmid, shuttle plasmid, and packaging plasmid were packaged together to infect GP-1 cells before being prepared into viral particles.

[0102] Test items and results

[0103] (1) HE and Oil Red O staining of mouse liver

[0104] After feeding, the mice were dissected, and liver samples were taken for HE staining and Oil Red O staining analysis. The specific procedures are as follows.

[0105] For HE staining analysis: mouse liver samples were fixed with 4% paraformaldehyde for 24 h, dehydrated and paraffin-impregnated, and then embedded in an embedding machine and sectioned (4 μm thick paraffin sections). The sections were stained with hematoxylin and eosin, dehydrated and mounted, and images were acquired and analyzed under a microscope.

[0106] When performing Oil Red O staining on liver samples: liver tissue was first removed from the body and fixed with 4% paraformaldehyde for 24 h, embedded in OCT and incubated at -80°C overnight, frozen sections (4 μm thick), washed with PBS, and then stained with Oil Red O using the Oil Red O staining kit (C0157S, Beyotime) according to the instructions.

[0107] Experimental results are as follows Figure 3 ( Figure 3 As shown in D), the analysis shows that: HE staining results indicate that compared with other treatment groups, the hepatocytes in the gene-silenced AAV8-shFer1l6 group are neatly arranged, and lipid deposition, cell edema, and inflammatory cell infiltration are significantly reduced; Oil Red O staining results indicate that compared with other treatment groups, the lipid droplets in the hepatocytes of the gene-silenced AAV8-shFer1l6 group are significantly reduced, and the lipid droplet volume is relatively small.

[0108] (2) Expression of FER1L6 and GLUT4

[0109] The expression levels of FER1L6 and GLUT4 proteins were detected and analyzed using Western blot technology. The specific procedure is as follows:

[0110] The pulverized liver tissue was lysed with lysis buffer (RIPA cell lysis buffer: protease phosphatase inhibitor: PMSF protease inhibitor = 100:1:1), and the lysis products were centrifuged at 4°C and 12000 r / min for 10 min, and the supernatant was collected.

[0111] After determining the protein concentration in the supernatant using the BCA protein quantification kit, 5× protein loading buffer was added to dilute the buffer to 1×. After high-temperature denaturation at 100°C, samples (protein samples) were taken for separation by 10% SDS-polypropylene gel electrophoresis.

[0112] Next, the proteins separated by electrophoresis gel were transferred to a polyvinylidene fluoride (PVDF) membrane and blocked with 5% skim milk powder for 2 h.

[0113] The primary antibody (FER1L6, GLUT4 and β-actin) was diluted with TBST (TBS: Tween 20 = 200:1) at a ratio of 1000:1 to form the primary antibody solution, which was then incubated with a PVDF membrane overnight at 4°C.

[0114] After incubation, wash the membrane 5 times with TBST (5 min each time);

[0115] Next, the horseradish peroxidase-labeled secondary antibody was diluted with TBST at a ratio of 5000:1 to prepare a secondary antibody solution, and then hybridized with the PVDF membrane washed with TBST as described above. The membrane was incubated at room temperature for 2 h. After incubation, the membrane was washed with TBST 5 times (5 min each time).

[0116] Finally, FER1L6, GLUT4 and β-actin proteins were visualized using ECL luminescent solution.

[0117] Experimental results are as follows Figure 3 ( Figure 3 As shown in C), the analysis shows that knocking down the Fer1l6 gene in mouse liver significantly reduced the Fer1l6 protein content, but significantly increased the GLUT4 protein content.

[0118] (3) mRNA expression of FER1L6 and GLUT4

[0119] The mRNA expression of FER1L6 and GLUT4 was detected using real-time quantitative PCR. The specific procedure is as follows:

[0120] First, total RNA was extracted from liver samples using Trizol Reagent reagent, and then the RNA was reverse transcribed into cDNA using the PrimeScript™ RT reagent kit with gDNA Eraser kit.

[0121] Subsequently, using the above cDNA as a template, a PCR reaction was performed with reference to the SYBR® Green Pro Taq HS premixed qPCR kit;

[0122] During the reaction, β-actin was used as an internal reference; the relevant reaction system is described above.

[0123] The reaction conditions were: 95°C pre-denaturation for 3 min; 95°C for 5 s, 60°C for 30 s, 40 cycles; the relative expression level of the gene was calculated using the double Δ method based on the cyclethreshold (Ct value).

[0124] The results are as follows Figure 3 ( Figure 3 A, Figure 3 As shown in B and 3C), the analysis shows that while the expression level of Fer1l6 protein in mouse liver was significantly decreased (P<0.001), the gene expression level of GLUT4 was significantly increased (P<0.05).

[0125] Further analysis of serum triglyceride and total cholesterol levels in mice from different treatment groups showed that knocking down Fer1l6 significantly reduced serum triglyceride and total cholesterol levels in diabetic mice. Figure 3 E), and the levels of superoxide dismutase and malondialdehyde in the liver (E), Figure 3 F). This result indicates that inhibiting the Fer1l6 gene can improve diabetic phenotypes such as lipid metabolism and oxidative stress damage in diabetic mice.

Claims

1. The application of FER1L6 gene inhibitors in the preparation of drugs for the prevention and treatment of type 2 diabetes mellitus (T2DM), characterized in that, The inhibitor increases GLUT4 expression by inhibiting FER1L6 expression, thereby improving the physiological metabolism of diabetic organisms; the physiological metabolism includes glucose uptake, lipid metabolism, and oxidative stress damage. The inhibitor inhibits the transcription and expression of FER1L6 by implementing lentiviral infection; the target sequence is shown in SEQ ID No.1, specifically: CCGGAAGATTGGAGATAAA.

2. The application of the FER1L6 gene inhibitor as described in claim 1 in the preparation of a drug for the prevention and treatment of type 2 diabetes mellitus (T2DM), characterized in that, The lipid metabolism includes triglycerides (TG) and total cholesterol (TC).

3. The application of the FER1L6 gene inhibitor as described in claim 1 in the preparation of a drug for the prevention and treatment of type 2 diabetes mellitus (T2DM), characterized in that, The indicators of oxidative stress damage include superoxide dismutase (SOD) and malondialdehyde (MDA).

4. A drug for the prevention and treatment of type 2 diabetes mellitus (T2DM), characterized in that, The preventive agent contains an inhibitor of the FER1L6 gene; the inhibitor inhibits the transcription and expression of FER1L6 by implementing lentiviral infection; the target sequence is shown in SEQ ID No.1, specifically: CCGGAAGATTGGAGATAAA.

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