Composition for targeted regulation of epigenetic modification level of type 2 diabetes mellitus gene

Abstract

The invention belongs to the technical field of biology, and discloses a composition for targeted regulation of type 2 diabetes gene epigenetic modification level, which comprises a formula A and a formula B. The formula A comprises the following components in percentage by mass: 86.5-90% of inulin, 9-12% of dihydroquercetin and 1-1.5% of vitamin C; and the formula B comprises the following components in percentage by weight: 80 to 88 percent of fructo-oligosaccharide, 5 to 15 percent of honeysuckle flower powder, 1 to 7 percent of betaine, 0.1 to 0.3 percent of vitamin C, 0.001 to 0.002 percent of vitamin B2, 0.001 to 0.002 percent of vitamin B6, 0.000004 to 0.000008 percent of vitamin B12 and 0.00004 to 0.00006 percent of folic acid. By designing the components and proportions of the formula A and the formula B and by means of nutrition intervention, part of the components serve as nutrient substances to participate in one-carbon unit metabolism, it is found in practice that the DNA methylation level of a TGFB1 gene expression regulation region can be regulated, high expression of TGFB1 is inhibited, the risk of suffering from type 2 diabetes mellitus and type 2 diabetic nephropathy is reduced, and the application prospect is wide. Type 2 diabetes and complications are improved and relieved, and the effects of preventing, improving and treating the type 2 diabetes are achieved.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and specifically relates to a composition that targets and regulates the epigenetic modification level of genes in type 2 diabetes. Background Technology

[0002] Type 2 diabetes is a common chronic metabolic disease with a complex pathogenesis, primarily related to insulin resistance and relative insulin insufficiency. Insulin resistance means that the body's cells are not sensitive to insulin, preventing glucose from effectively entering cells for utilization and leading to elevated blood sugar. As the disease progresses, the function of pancreatic beta cells gradually declines, and insulin secretion also gradually decreases. Symptoms of type 2 diabetes may include polydipsia, polyphagia, polyuria, and weight loss, but some patients may not have obvious symptoms in the early stages and are often diagnosed during physical examinations or when complications arise. Treatment typically includes lifestyle interventions, blood glucose monitoring, and medication.

[0003] DNA methylation refers to the process by which a methyl group on S-adenosylmethionine is transferred to the 5-carbon terminus of a cytosine residue in CpG, forming 5'-methylcytosine, under the action of DNA methyltransferases. DNA methylation occurs in tissues and organs associated with the pathogenesis of type 2 diabetes, such as pancreatic islet cells, adipose tissue, liver, and kidneys, and regulates the occurrence and evolution of type 2 diabetes genes. Therefore, existing technologies use DNA methylation as a biomarker to assess the risk of type 2 diabetes. Based on the above, it is necessary to research and develop a nutritional formula that can prevent and treat type 2 diabetes by regulating the methylation level of type 2 diabetes genes. Summary of the Invention

[0004] The purpose of this invention is to provide a composition that targets and regulates the epigenetic modification level of type 2 diabetes genes, and has the effects of regulating DNA methylation level, affecting blood glucose, and preventing and treating type 2 diabetes and its complications.

[0005] The above-mentioned technical objective of the present invention is achieved through the following technical solution: a composition for targeting and regulating the epigenetic modification level of type 2 diabetes genes, comprising formulation A and formulation B, wherein, by mass percentage, formulation A comprises 86.5-90% inulin, 9-12% dihydroquercetin, and 1-1.5% vitamin C; and formulation B comprises 80-88% fructooligosaccharides, 5-15% honeysuckle powder, 1-7% betaine, 0.1-0.3% vitamin C, 0.001-0.002% vitamin B2, 0.001-0.002% vitamin B6, 0.000004-0.000008% vitamin B12, and 0.00004-0.00006% folic acid.

[0006] By adopting the above technical solutions, the nutritional formulation of the present invention includes formulation A and formulation B.

[0007] In Formula A, inulin is a natural functional dietary fiber, belonging to the plant polysaccharide family. Inulin is not broken down and absorbed during ingestion, but it can produce glycogenic fatty acids in the intestines, which can inhibit gluconeogenesis and enhance insulin resistance. Existing research has shown that inulin can improve diseases such as hyperglycemia caused by glucose and lipid metabolism disorders, and has a regulatory effect on glucose and lipid metabolism, but the regulatory mechanism is still unclear.

[0008] Dihydroquercetin, also known as taxane, is a naturally occurring dihydroflavonol compound and an antioxidant. It possesses hepatoprotective and nephroprotective effects, as well as anti-inflammatory, antibacterial, enzyme-regulating, and microcirculation-improving properties. Current research has shown that high-dose dihydroquercetin treatment of diabetic nephropathy rats can reduce kidney tissue pathological damage and the secretion of inflammatory factors, thus exerting a nephroprotective effect; however, it has not been applied to humans.

[0009] In Formula B, fructooligosaccharides, also known as fructooligosaccharides or sucrose trisaccharide oligosaccharides, are found in many commonly consumed fruits and vegetables. Fructooligosaccharides possess both definite health benefits and excellent food ingredient qualities. They are characterized by their low calorie content, non-caries properties, promotion of bifidobacteria growth, blood sugar reduction, improvement of serum lipids, and promotion of trace element absorption. In recent years, among many oligosaccharide-based foods, fructooligosaccharides have been recognized by international nutritionists as "a water-soluble dietary fiber with excellent indigestibility," capable of bidirectionally regulating the human microecological balance, and are a typical super-strong bifidogenic factor.

[0010] Honeysuckle powder refers to the powder made from the dried flower buds or unopened flowers of *Lonicera japonica*, a plant belonging to the genus *Lonicera* in the family Caprifoliaceae. Its main components include chlorogenic acid and luteolin. In terms of efficacy, it has heat-clearing and detoxifying effects, and can improve symptoms such as sore throat and dysentery to some extent; it also helps improve liver cell activity, enhances the liver's detoxification function, and has a certain protective effect on the liver.

[0011] Betaine is an important nutrient obtained from various foods. It is rapidly absorbed and used as a source of osmotic pressure and methyl groups, thus helping to maintain the health of the liver, heart, and kidneys. Betaine can reduce elevated serum homocysteine ​​levels associated with mild or severe hyperhomocysteinuria through the methionine cycle and plays a role in epigenetics and athletic performance. Betaine is a quaternary ammonium-type water-soluble alkaloid that can act as a methyl donor, serving as a substitute for methionine and choline chloride. It also relieves stress, regulates osmotic pressure, improves appetite, and reduces fat. Current technologies utilize betaine hydrochloride for the prevention and treatment of atherosclerosis, hepatobiliary diseases, and rheumatic diseases. Existing research in animal experiments has shown that betaine has hypoglycemic, renal damage-improving, hepatoprotective, and lipid-lowering functions in type 2 diabetic mouse models.

[0012] Methylation risk scoring is a method for risk assessment using DNA methylation characteristics. RNA sequence analysis is performed on islet samples with methylation data from type 2 diabetes patients and controls, focusing only on protein-coding genes. Differentially expressed genes are identified in the islets of patients and controls, and a weighted composite methylation risk score (MRS) is calculated for each gene. These genes show altered expression and contain five or more distinct methylation CpG sites within or near the gene, allowing for the study of the combined effect of multiple different methylation sites within a single gene. A higher methylation risk score is associated with a higher risk of developing type 2 diabetes.

[0013] Based on this, the present invention innovatively adopts the formulation design of Formula A and Formula B, aiming to regulate the methylation level of diabetes-related genes through nutritional intervention, thereby reducing the risk of type 2 diabetes, alleviating and improving type 2 diabetes and its complications, and playing a role in the prevention and treatment of type 2 diabetes.

[0014] In one-carbon metabolism, S-adenosylmethionine is the sole methyl donor for many biological methylation reactions, including DNA and histone methylation. S-adenosyl-L-homocysteine ​​(SAH) is an inhibitor of methyltransferases such as DNA methyltransferases and histone methyltransferases. In single-carbon metabolism, vitamins B2, B6, B12, and folic acid are coenzymes, while methionine, choline, betaine, and serine are methyl donors.

[0015] This invention designs formulas A and B. In formula A, inulin and dihydroquercetin regulate glucose and lipid metabolism after ingestion. Nutrients involved in one-carbon unit metabolism, such as vitamin B2, vitamin B6, vitamin B12, and folic acid, participate in DNA methylation by regulating the levels of the universal methyl donor S-adenosylmethionine and the methyltransferase inhibitor S-adenosylhomocysteine. Through the design of the various components and proportions in formulas A and B, this invention can regulate DNA methylation levels after ingestion by the human body. Practical experience has shown that it can effectively regulate DNA methylation in the TGFB1 gene expression regulatory region. In type 2 diabetes patients, diabetic nephropathy is a common complication. DNA methylation in the TGFB1 gene expression regulatory region leads to the activation of TGFB1 mRNA expression, resulting in high TGFB1 expression, causing extracellular matrix accumulation, and leading to glomerular sclerosis. Formulas A and B of this invention can effectively regulate DNA methylation in the TGFB1 gene expression regulatory region and inhibit high TGFB1 expression, thus playing a very effective role in the prevention and treatment of type 2 diabetic nephropathy.

[0016] While there are precedents in existing technologies for studying the effects of betaine and dihydroquercetin on type 2 diabetic mice in animal experiments, it is important to note that: First, the mice used in animal experiments may differ from humans in organ structure, physiological function, and metabolic pathways, leading to differences in the absorption, distribution, and metabolism of ingested substances. Second, the human immune system is more complex than that of animal models, and its response to drugs may differ, potentially resulting in variations in drug efficacy and side effects. Third, the manifestations and pathophysiological mechanisms of diseases may not be entirely identical between animal models and humans. Therefore, studies in animal models cannot directly infer that the drugs will necessarily be effective in humans.

[0017] In this invention, through two sets of formulations and 10 components and proportions of inulin, dihydroquercetin, fructooligosaccharides, honeysuckle powder, betaine, vitamin C, vitamin B2, vitamin B6, vitamin B12, and folic acid, effective nutritional intervention can be achieved in the human body. By regulating the level of DNA methylation, a significant inhibitory effect on the high expression of TGFB1 is achieved, which plays a role in the prevention, improvement and treatment of type 2 diabetes and type 2 diabetic nephropathy.

[0018] A further provision of the present invention is that, by weight percentage, formulation A comprises 90% inulin, 9% dihydroquercetin, and 1% vitamin C.

[0019] A further provision of the present invention is that, by weight percentage, formulation B comprises 85% fructooligosaccharides, 10% honeysuckle powder, 4% betaine, 0.2% vitamin C, 0.0015% vitamin B2, 0.0015% vitamin B6, 0.000006% vitamin B12, and 0.00005% folic acid.

[0020] A further provision of the present invention is that formulation A and formulation B are taken 8-14 hours apart.

[0021] By adopting the above technical solution, during nutritional intervention, Formula A and Formula B are taken once in the morning and once in the evening.

[0022] A further setting of the present invention is that the mass ratio of formulation A to formulation B is 1:1.

[0023] A further setting of the present invention is that the dosage of formula A and formula B is one packet per day, and each packet contains 10g.

[0024] The beneficial effects of this invention are:

[0025] 1. This invention designs the components and proportions of Formula A and Formula B, and through nutritional intervention, some components participate in one-carbon unit metabolism as nutrients. In practice, it has been found that this can regulate the DNA methylation level in the TGFB1 gene expression regulatory region, inhibit the high expression of TGFB1, reduce the risk of type 2 diabetes and type 2 diabetic nephropathy, improve and alleviate type 2 diabetes and type 2 diabetic nephropathy, and play a role in the prevention, improvement and treatment of type 2 diabetes.

[0026] 2. This invention, through the selection and optimization of components and proportions, dosage, and dosing intervals, can effectively regulate the methylation of certain type 2 diabetes-related genes in the human body, such as the TGFB1 gene expression regulatory region, without side effects and is safe and reliable. For healthy individuals, this can reduce the risk of developing type 2 diabetes and type 2 diabetic nephropathy. Furthermore, this nutritional formula can also be used as a specific lifestyle intervention in the treatment of type 2 diabetes. Detailed Implementation

[0027] The technical solutions in the embodiments will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0028] A composition targeting and regulating the epigenetic modification level of genes in type 2 diabetes includes Formula A and Formula B. Each 100g of Formula A contains 90g inulin, 9g dihydroquercetin, and 1g vitamin C. Each 100g of Formula B contains 85g fructooligosaccharides, 10g honeysuckle powder, 4g betaine, 200mg vitamin C, 1.5mg vitamin B2, 1.5mg vitamin B6, 6ug vitamin B12, and 50ug folic acid. Dosage: One 10g packet of Formula A in the morning and one 10g packet of Formula B in the evening, 12 hours apart.

[0029] Experimental Section: Sulfite sequencing is a common method for detecting DNA methylation. It converts unmethylated cytosine into uracil, and then analyzes the methylation level through sequencing. The basic principle of sulfite sequencing is to treat DNA with sulfite, converting unmethylated cytosine into uracil, while methylated cytosine remains unchanged. After PCR amplification, sequencing is performed; unmethylated cytosine appears as thymine during sequencing.

[0030] Ninety-one patients with type 2 diabetes were selected and divided into two groups: one receiving nutritional intervention (DM group) and the other receiving no nutritional intervention (CON group). Genomic DNA was extracted from peripheral blood of all subjects and modified with sodium bisulfite. Methylation-specific PCR was used to initially screen for DNA methylation in the TGFFB1 gene expression regulatory region of each group. After bisulfite modification, sequencing was used to detect the DNA methylation level in the TGFFB1 gene expression regulatory region of each group.

[0031] After two months of nutritional intervention, the DNA methylation rate in the TGFB1 gene expression regulatory region of the DM group was 18.6%, higher than the 12.2% in the CON group. Nutritional intervention increased the methylation level of specific gene promoter regions in the patients' blood and reduced the expression level of the TGFB1 gene, which is beneficial to the patients' health.

Claims

1. A composition for targeted modulation of the level of epigenetic modification of a type 2 diabetes gene, characterized in that: The formula A, the formula B, by mass percentage, the formula A includes 86.5-90% inulin, 9-12% dihydroquercetin, 1-1.5% vitamin C;The formula B includes 80-88% fructooligosaccharide, 5-15% honeysuckle powder, 1-7% betaine, 0.1-0.3% vitamin C, 0.001-0.002% vitamin B2, 0.001-0.002% vitamin B6, 0.000004-0.000008% vitamin B12, 0.00004-0.00006% folic acid.

2. The composition for targeting and modulating the level of epigenetic modification of a type 2 diabetes gene according to claim 1, wherein: The formula A includes 90% inulin, 9% dihydroquercetin, 1% vitamin C by mass percentage.

3. The composition for targeting and modulating the level of epigenetic modification of a type 2 diabetes gene according to claim 1 or 2, characterized in that: The formula B includes 85% fructooligosaccharide, 10% honeysuckle powder, 4% betaine, 0.2% vitamin C, 0.0015% vitamin B2, 0.0015% vitamin B6, 0.000006% vitamin B12, 0.00005% folic acid by mass percentage.

4. The composition for targeting and modulating the level of epigenetic modification of a type 2 diabetes gene according to claim 1, wherein: The mass ratio of the formula A and the formula B is 1:

1.

5. The composition for targeting and modulating the level of epigenetic modification of a type 2 diabetes gene according to claim 1, wherein: The formula A and the formula B are taken with an interval of 8-14h.

6. The composition for targeting and modulating the level of epigenetic modification of a type 2 diabetes gene according to claim 1, wherein: The formula A, the formula B are taken one package per day, each containing 10g.