Application of verbena glycoside in treating diabetes and / or diabetic complications

Abstract

The present invention provides the use of verbena glycoside in the preparation of a drug for treating diabetes, the use of verbena glycoside in the preparation of a drug for treating diabetic heart disease, and the use of verbena glycoside in the preparation of a drug for treating diabetic nephropathy. The present invention has found for the first time through research that verbena glycoside has the effect of preventing and treating diabetes and diabetic heart and kidney damage: in terms of blood sugar, verbena glycoside can improve the glucose tolerance level by improving the blood sugar metabolism level; in terms of heart damage caused by diabetes, verbena glycoside can improve cardiac dysfunction and left ventricular remodeling, and inhibit the fibrosis of heart tissue; in terms of kidney damage, verbena glycoside can improve the level of kidney function and inhibit the level of kidney fibrosis. The present invention provides a strong experimental basis and theoretical basis for the use of verbena glycoside in the prevention and treatment of diabetes and its heart and kidney damage, and broadens its application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine, and specifically relates to the application of verbena glycoside in treating diabetes and / or diabetic complications. Background Art

[0002] Diabetes is a group of lifelong metabolic diseases caused by multiple causes and characterized by persistent chronic hyperglycemia. Diabetes has become an increasingly serious health problem. Diabetes is a disease with the most known complications. Most diabetic patients die from diabetic complications. Long-term high blood sugar levels damage large and microvascular vessels and endanger the heart, kidneys, peripheral nerves, eyes, feet, etc. At present, diabetic heart disease and diabetic nephropathy are the main causes of death in diabetic patients.

[0003] Clinically, the treatment options for patients with diabetes and cardiorenal complications are very limited, mainly including active treatment of diabetes as the primary disease, strict control of blood sugar, dietary therapy, and the use of insulin and other hypoglycemic drugs. However, there is still a lack of effective therapeutic drugs to protect diabetes and the heart and kidney damage caused by it. By intervening in the high sugar state and heart and kidney damage caused by the disease, the threat of diabetes to human health can be reduced to a certain extent. As a result, the treatment of diabetes and its cardiorenal complications has received increasing attention and has become an urgent problem to be solved in the field of heart and kidney related diseases.

[0004] At present, there are many oral drugs for the prevention and treatment of diabetes in the domestic and foreign markets, and common diabetes treatment drugs can be divided into eight categories. Mainly including insulin and its analogs, metformin compounds, sulfonylurea drugs, glucosidase inhibitors, thiazolidinedione drug sensitizers, DPP-4 enzyme inhibitors and receptor agonists, etc. These drugs have played a great role in the treatment and prevention of diabetes and the relief of the suffering of diabetics. However, there are many varieties of diabetes treatment drugs currently available on the market, most of which are biological products or artificially synthesized monomer compounds, and there are also their own side effects. And with the diversification of complications of diabetics, especially cardiorenal damage, human health is seriously threatened. Therefore, the development of safer and more effective drugs for the treatment of diabetes (hyperglycemia) and its cardiorenal damage is still a very meaningful work.

[0005] Currently, many traditional Chinese medicines have been widely used in the prevention and treatment of diabetes and its complications. Traditional Chinese medicines exert their pharmacodynamic effects through multiple pathways, multiple targets, and integrated regulatory mechanisms, and have systemic characteristics. However, the chemical composition of traditional Chinese medicines is complex, the material basis of the pharmacological effects is unclear, and it is difficult to control the quality. The efficacy and emphasis of various effective parts of traditional Chinese medicines used to treat diabetes and its complications are different. Replacing traditional Chinese medicines with effective chemical components of traditional Chinese medicines not only facilitates the quality control of drugs, but also reduces their complexity. Therefore, providing more convenient and effective effective chemical components of traditional Chinese medicines has important clinical significance. Summary of the invention

[0006] Based on this, the purpose of the present invention is to provide the use of verbena glycoside in the treatment of diabetes and / or diabetic complications. Verbena glycoside has the effect of preventing and treating diabetes and diabetic heart and kidney damage, and can be used to prepare drugs for the treatment of diabetes, diabetic heart disease and diabetic nephropathy.

[0007] To achieve the above object, the present invention adopts the following technical solution.

[0008] Application of verbena glycoside in preparing medicine for treating diabetes.

[0009] In some embodiments, the drug has at least one of the following effects: lowering blood sugar content, improving glucose tolerance level.

[0010] The present invention also provides the use of verbena glycoside in preparing medicine for treating diabetic heart disease.

[0011] In some embodiments, the drug has at least one of the following effects: improving cardiac dysfunction, left ventricular remodeling, and improving cardiac fibrosis.

[0012] The present invention also provides the use of verbena glycoside in preparing medicine for treating diabetic nephropathy.

[0013] In some embodiments, the drug has at least one of the following effects: improving renal function level, protecting renal structure, and improving renal fibrosis.

[0014] The present invention also provides a medicine for treating diabetes and / or diabetic heart disease and / or diabetic nephropathy, wherein the medicine comprises verbena glycoside and pharmaceutically acceptable excipients.

[0015] In some embodiments, the drug is in the form of a solution, granules, tablets, pills, drops, injections, capsules, powders or decoctions.

[0016] The present invention has found for the first time through research that verbena glycoside has the effect of preventing and treating diabetes and diabetic heart and kidney damage: in terms of blood sugar, verbena glycoside can improve the glucose tolerance level by improving the blood sugar metabolism level; in terms of heart damage caused by diabetes, verbena glycoside can improve cardiac dysfunction and left ventricular remodeling, and inhibit the fibrosis of cardiac tissue; in terms of kidney damage, verbena glycoside can improve the level of renal function and inhibit the level of kidney fibrosis. Verbena glycoside has the advantages of stable hypoglycemic effect, taking into account complications, definite efficacy, clear composition, no obvious toxic and side effects, convenient to take and low price. Compared with the existing traditional Chinese medicine for treating diabetes and complications, its chemical composition is simple and clear, it is easier to explain its mechanism of action in pharmacological research, and it is easier to control the quality of drugs in production. The present invention provides a strong experimental basis and theoretical basis for the use of verbena glycoside in the prevention and treatment of diabetes and its heart and kidney damage, and broadens its application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The experimental results of verbena glycoside improving the general condition and blood sugar level of diabetic mice; A is the weight test result of mice; B is the fasting blood sugar level test result of mice; C is the result of oral glucose tolerance test of mice.

[0018] Figure 2 These are the experimental results showing that verbena glycoside improves cardiac dysfunction, left ventricular remodeling and cardiac fibrosis in diabetic model mice; A is the echocardiography result; B is the heart WGA staining result; and C is the heart Sirius red staining result.

[0019] Figure 3 The figures show the experimental results of verbena glycoside improving the renal function and renal fibrosis of diabetic mice; A is the creatinine (SCr) level test result of diabetic mice; B is the kidney PAS staining result; C is the kidney Masson staining result. DETAILED DESCRIPTION

[0020] The experimental methods in the following examples of the present invention, where no specific conditions are specified, are usually carried out under conventional conditions or under conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0021] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0022] The terms "including" and "having" and any variations thereof of the present invention are intended to cover non-exclusive inclusions. For example, a process, method, device, product or equipment comprising a series of steps is not limited to the listed steps or modules, but may optionally include steps not listed, or may optionally include other steps inherent to these processes, methods, products or equipment.

[0023] In the present invention, the term "multiple" refers to two or more than two. "And / or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can represent the following three situations: A exists alone, A and B exist at the same time, and B exists alone. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0024] The following describes the invention in conjunction with specific embodiments.

[0025] Example 1

[0026] 1. Experimental Methods

[0027] 1. Medication

[0028] Verbena glycoside standard was purchased from Chengdu Mansite Biotechnology Co., Ltd.

[0029] 2. Animal Model Preparation

[0030] SPF C57 / BL male mice (35-42 days old, weighing 16-20g) were randomly divided into a normal group (Normal group) and a diabetic group (DN group). Mice in the DN group were fed a high-fat diet, and mice in the Normal group were fed a normal diet. Two months later, all mice received intraperitoneal injections of sodium citrate (pH 4.5, NC group) or 50mg / kg STZ (dissolved in sodium citrate, pH 4.5, DN group) for 5 consecutive days. One week later, mice in the DN group with fasting blood glucose levels higher than 16.7mmol / L were used for subsequent experiments. After successful modeling, the DN group was divided into two groups: the model group (Model group) and the group treated with 50mg / kg verbena glycoside by gavage (Cornin group). Gavage was performed once a day, with a volume of 0.2mL each time; among them, the Normal group and the Model group were gavaged with an equal volume of normal saline. The gavage lasted for 4 weeks. During the entire experiment, the overall condition of each group of mice was observed every morning and evening, including general condition, mental and hair condition, defecation and urination, and changes in food and water intake.

[0031] 3. General indicators

[0032] During the administration process, the mice's eating, drinking, and excretion changes were carefully observed every day, and the general conditions such as mental state, hair changes, and death of each group of mice were compared, and weight changes and deaths were recorded.

[0033] 4. Blood sugar level measurement

[0034] All experimental mice had their fasting blood glucose measured after fasting for 12 hours each week, and the weight and fasting blood glucose levels of the four groups of mice were observed. After the experiment, an oral glucose tolerance test (OGTT) was performed on mice to measure the blood glucose levels at 30, 60, 90 and 120 minutes after gavage, and the area under the blood glucose curve within 120 minutes was calculated using Graphpad. 24 hours after the last administration of each group of mice, the eyeballs were removed for blood sampling, and serum was taken to test relevant biochemical indicators.

[0035] 5. Echocardiography

[0036] After intragastric administration, the cardiac function of mice was evaluated by small animal ultrasound. The short-axis view at the level of the papillary muscles of mice, cardiac contractile function index including left ventricular ejection fraction (LVEF) and left ventricular short axis shortening rate (LVFS) were measured and quantified in each test. The above parameters were analyzed using relevant modules.

[0037] 6. Measurement of renal function biochemical indicators

[0038] After intragastric administration, the renal function of the mice was evaluated by the detection kit. Before the experimental animals were taken, 5% chloral hydrate 10mL / kg was injected for anesthesia, and blood was collected by eyeball blood collection method, kept at room temperature for 4 hours, centrifuged at 4℃3000r·min-1 for 15min, and the supernatant was collected and stored at -80℃ for the determination of urea nitrogen (SCr), an indicator of mouse renal function.

[0039] 7. Heart and kidney pathology

[0040] After 4 weeks of continuous administration, all mice were over-anesthetized, the heart and kidneys of the mice were freed, and the heart and one kidney were placed in a 4% paraformaldehyde solution as a fixative. After fixation, they were embedded in paraffin and the Sirius red staining and WGA staining of heart tissue, and PAS staining and Masson staining of kidney tissue were performed.

[0041] 2. Experimental Results

[0042] 1. Verbena glycoside improves the general condition of mice

[0043] After modeling, the general condition of the mice was observed. The Normal group mice had shiny and smooth fur, good mental state, free movement, continuous weight gain, normal food intake, water intake, and defecation and urination. The litter was dry and had no obvious odor. The fur of the diabetic mice in the Model and Cornin groups gradually became yellow, dull, and easy to fall off. They were in poor spirits, less active, and sleepy. They were thin and lost weight. They had typical symptoms of polyphagia, polydipsia, and polyuria. The litter was easily damp and had an obvious odor. After 4 weeks of administration, the mental state of the mice in the Cornin group improved, their fur gradually became clean and shiny, and their food intake, water intake, and excretion decreased. The weight of mice in each group changed during the administration period. Except for the weight gain of the Normal group mice, the weight loss of the Model group mice was the largest, and the weight loss of the Cornin group mice was smaller. Compared with the Model group, the difference was statistically significant (P < 0.01) ( Figure 1 A). The results suggest that verbena glycoside treatment can improve the general condition of diabetes, relieve the three more and one less symptoms of diabetic mice, and has a certain protective effect on weight loss in diabetic mice.

[0044] 2. Verbena glycoside improves fasting blood glucose (FBG) and glucose tolerance (OGTT) levels in diabetic model mice

[0045] The fasting blood glucose level (FBG) of mice induced by STZ was significantly increased (P < 0.0001), while verbena glycosides could significantly reduce the blood glucose level, and the difference was statistically significant (P < 0.0001) ( Figure 1 B), indicating that verbena glycosides can effectively regulate and improve blood sugar levels.

[0046] According to the results of the oral glucose tolerance test (OGTT) in mice and the statistical analysis of the area under the curve by t-test, the glucose tolerance of the Model group was severely impaired, and the blood glucose level at 0 min was significantly higher than that of the other groups, and it increased rapidly within 30 min. The AUC of the Cornin group was significantly lower than that of the Model group, and the difference was statistically significant (P < 0.0001) ( Figure 1 C), indicating that verbena glycosides can improve the glucose tolerance level when administered orally, and may thereby play a role in treating diabetes.

[0047] 3. Verbena glycoside improves cardiac dysfunction, left ventricular remodeling and cardiac fibrosis in diabetic mice

[0048] Echocardiography showed that STZ-induced diabetic mice could cause cardiac contractile dysfunction, and verbena glycosides could improve the LVEF value level (P<0.05) and LVFS value level (P<0.01) of diabetic model mice ( Figure 2 A).

[0049] Cardiac WGA staining showed that STZ-induced diabetic mice can cause myocardial hypertrophy. Myocardial hypertrophy of diabetic mice treated with verbena glycosides for 4 weeks was significantly improved, which can effectively inhibit myocardial hypertrophy and significantly reduce the cross-sectional area of ​​myocardial cells ( Figure 2 B).

[0050] Sirius red staining of the heart showed that the level of myocardial fibrosis in STZ-induced diabetic mice was significantly increased, and verbena glycosides could effectively improve cardiac fibrosis ( Figure 2 C).

[0051] 4. Verbena glycoside improves renal function and renal fibrosis in diabetic mice

[0052] like Figure 3 A shows that the creatinine (SCr) level of STZ-induced diabetic mice was significantly increased, indicating that the renal function of STZ-induced diabetic mice was impaired. The intervention of verbena glycosides can improve the renal function level, and the difference is statistically significant (P < 0.01).

[0053] Kidney PAS staining showed that STZ-induced diabetic mice had vacuolar degeneration of renal tubular epithelial cells, and the widening of the renal mesangial area was more significant than that of the normal group. Some renal tubules dilated in the renal interstitium, and renal tubular epithelial cells shed and regenerated. The above changes were significantly alleviated after verbena glycoside treatment. It can be seen that verbena glycoside has a significant protective effect on the structure of the kidneys of diabetic nephropathy mice ( Figure 3 B).

[0054] Renal Masson staining showed that glomerular collagen deposition was clearly visible in STZ-induced diabetic mice. Verbena glycosides can significantly improve this situation. Such results indicate that the level of renal fibrosis in STZ-induced diabetic mice is aggravated, and the intervention of verbena glycosides can effectively improve the level of renal fibrosis ( Figure 3 C).

[0055] In summary, verbena glycosides can improve glucose tolerance by improving blood sugar metabolism. In terms of heart damage caused by diabetes, verbena glycosides can improve cardiac dysfunction and left ventricular remodeling, and inhibit cardiac tissue fibrosis; in terms of kidney damage, verbena glycosides can improve kidney function and inhibit kidney fibrosis, thus playing a therapeutic role in diabetes and its heart and kidney damage.

[0056] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0057] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. Use of verbena glycoside as the sole active ingredient in the preparation of a drug for treating diabetes, diabetic heart disease or diabetic nephropathy; The drug has at least one of the following effects: Lower blood sugar levels and improve glucose tolerance levels; Improve cardiac dysfunction, left ventricular remodeling and cardiac fibrosis; Improve renal function levels, protect renal structure and improve renal fibrosis.

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