Application of rare ginseng component T19 in preparation of products for preventing and / or treating glycolipid metabolic disorder

Abstract

The present application relates to a kind of ginseng rare component T19 in preparation of the application of preventing and / or treating glycolipid metabolism disorder product, belong to biological medicine technical field.The present application first discovers that ginseng rare component T19 has good prevention and treatment effect to glycolipid metabolism disorder, ginseng rare component T19 shows good regulation glycolipid metabolism effect, in preparation preventing and / or treating glycolipid metabolism disorder product aspect has good application prospect, the product includes food, health care product and drug, this new application has great significance to break through glycolipid metabolism disorder prevention and / or treatment bottleneck.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to the application of a rare ginseng ingredient, T19, in the preparation of products for the prevention and / or treatment of disorders of glucose and lipid metabolism. Background Technology

[0002] Glycolipid metabolism disorder is a metabolic imbalance caused by multiple factors (such as genetics, obesity, poor diet, and lack of exercise). It is mainly manifested as elevated blood glucose, dyslipidemia (elevated total cholesterol and triglycerides, decreased high-density lipoprotein, etc.), and insulin resistance. Further development can lead to various complications such as type 2 diabetes, atherosclerosis, fatty liver, and cardiovascular disease, seriously threatening human health. Currently used drugs for regulating glucose and lipid metabolism have significant side effects and diminishing efficacy with long-term use. Therefore, developing safe and effective natural active ingredients for the prevention and treatment of glucose and lipid metabolism disorders has significant clinical value and market potential.

[0003] T19 belongs to the dammarane-type aglycone class of compounds, with the molecular formula C19. 30 H 54 O5 has the following structural formula:

[0004] ; T19 is a rare natural ginsenoside found in plants of the genus *Panax*, and is also one of the important active ingredients in ginseng, American ginseng, and other plants of the Araliaceae family. Current research has revealed that ginsenoside T19 possesses pharmacological activities such as antioxidant, anti-inflammatory, and cardiovascular protection; however, no existing technology has demonstrated that ginsenoside T19 regulates glucose and lipid metabolism or improves glucose and lipid metabolism disorders. Summary of the Invention

[0005] The purpose of this invention is to overcome the aforementioned deficiencies in the existing technology and provide an application of the rare ginseng component T19 in the preparation of products for the prevention and / or treatment of glucose and lipid metabolism disorders. Cell and animal experiments have shown that the rare ginseng component T19 exhibits good regulatory effects on glucose and lipid metabolism, demonstrating promising application prospects in the preparation of products for the prevention and / or treatment of glucose and lipid metabolism disorders. This is of great significance for overcoming the bottleneck in the prevention and / or treatment of glucose and lipid metabolism disorders.

[0006] Application of rare ginseng component T19 in the preparation of products for the prevention and / or treatment of disorders of glucose and lipid metabolism.

[0007] Furthermore, the effects of the product for preventing and / or treating disorders of glucose and lipid metabolism include at least one of the following: (a) Improve blood glucose and / or lipid metabolism, including lowering fasting blood glucose, improving glucose tolerance, and lowering serum total cholesterol and / or triglyceride levels; (b) Improve abnormal weight; (c) Protect the liver and improve liver damage, including reducing hepatic lipid accumulation and inhibiting the transcription of hepatic inflammatory factors; (d) Activation of insulin signaling pathways, including promoting insulin receptor phosphorylation, upregulating the expression or transcription of glucose transporter 4 (GLUT4), and / or upregulating the transcription and phosphorylation levels of AMP-activated protein kinase (AMPK); (e) Regulate the gut microbiota to normalize its structure.

[0008] Furthermore, the glucose and lipid metabolism disorders include hyperglycemia, hyperlipidemia, insulin resistance, and related metabolic abnormalities.

[0009] Preferably, the glucose and lipid metabolism disorder is diabetes.

[0010] Furthermore, the rare ginseng component T19 is a product obtained by hydrolyzing and recrystallizing the total saponins obtained by separating ginseng or American ginseng.

[0011] Furthermore, the concentration of the rare ginseng component T19 in the product is 20~60 mg / kg body weight.

[0012] Furthermore, the product is food, health supplement, or medicine.

[0013] Furthermore, the product includes the active ingredient ginseng rare component T19 and other excipients acceptable in food, health products or pharmaceuticals.

[0014] Furthermore, the product is available in oral and injectable dosage forms.

[0015] Furthermore, the oral dosage form is a tablet, capsule, granule, syrup, or emulsion.

[0016] Furthermore, the injection method for the injectable dosage form is subcutaneous injection, intramuscular injection, or intravenous injection.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention is the first to discover that the rare ginseng component T19 has good preventive and / or therapeutic effects on glucose and lipid metabolism disorders, and based on this, provides the application of the rare ginseng component T19 in the preparation of foods, health products, or pharmaceuticals for the prevention and / or treatment of glucose and lipid metabolism disorders. Animal experiments show that the rare ginseng component T19 can reduce fasting blood glucose and glycated hemoglobin levels in mice with glucose and lipid metabolism disorders, decrease serum total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels, increase high-density lipoprotein cholesterol levels, improve insulin resistance index, alleviate hepatic lipid deposition and steatosis, and dose-dependently regulate the expression of glucose and lipid metabolism-related proteins (such as IR, INS-1, AKT, AMPK, ACC, and GLUT4).

[0018] Therefore, the rare ginseng component T19 exhibits good regulatory effects on glucose and lipid metabolism and has promising application prospects in the preparation of foods, health products or medicines for the prevention and / or treatment of glucose and lipid metabolism disorders. The new application of the rare ginseng component T19 provided by this invention in the preparation of foods, health products or medicines for the prevention and / or treatment of glucose and lipid metabolism disorders is of great significance for breaking through the bottleneck in the prevention and / or treatment of glucose and lipid metabolism disorders. Attached Figure Description

[0019] Figure 1 The effect of T19 on body weight in STZ-induced diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0020] Figure 2 The effect of T19 on blood glucose in STZ-induced diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0021] Figure 3 The effect of T19 on glucose tolerance in STZ-induced diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0022] Figure 4 The effect of T19 on serum TG and TC levels in STZ-induced diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0023] Figure 5 The effect of T19 on body weight in DB / DB mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0024] Figure 6 The effect of T19 on blood glucose in DB / DB mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P<0.05 vs model group, P <0.01 vs model group).

[0025] Figure 7 The effect of T19 on glucose tolerance in DB / DB mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0026] Figure 8 The effect of T19 on serum TG and TC levels in DB / DB mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0027] Figure 9 The effect of T19 on body weight in HFD / STZ diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0028] Figure 10 The effect of T19 on blood glucose in HFD / STZ diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vsmodel group).

[0029] Figure 11 The effect of T19 on glucose tolerance in HFD / STZ diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vsmodel group).

[0030] Figure 12 The effect of T19 on insulin tolerance in HFD / STZ diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0031] Figure 13 The effects of T19 on TG, TC, LDL-C, and HDL-C in HFD / STZ diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0032] Figure 14 The effects of T19 on liver pathology in DB / DB mice were defined as follows: A was the blank control group (40X), A1 was 10X; B was the model group (DB / DB) (40X), B1 was 10X; C was the T19 group (40X), C1 was 10X; and D was the positive control group (rosiglitazone) (40X), D1 was 10X.

[0033] Figure 15 The effects of T19 on liver histopathological damage and liver glycogen in HFD / STZ mice are shown in Figure A, a photomicrograph of mouse liver tissue stained with H&E at 200x magnification; and Figure B, an assessment of liver glycogen accumulation by PAS staining at 200x magnification.

[0034] Figure 16 The effect of T19 on the expression of liver inflammatory factors in STZ-induced diabetic mice (Values ​​are expressed as mean ± SEM). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vsmodel group, P <0.01 vs model group).

[0035] Figure 17 The effect of T19 on the expression of liver inflammatory factors in DB / DB diabetic mice (Values ​​are expressed as mean ± SEM). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0036] Figure 18 The effect of T19 on GLUT4 and AMPK transcription levels in the skeletal muscle of STZ-induced diabetic mice (Values ​​are expressed as mean ± SD). P<0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vsmodel group, P <0.01 vs model group).

[0037] Figure 19 The effect of T19 on GLUT4 and AMPK transcription levels in skeletal muscle of DB / DB diabetic mice (Values ​​are expressed as mean ± SD). P <0.05 vs Ctrl group, P <0.01 vs Ctrl group, # P <0.05 vs model group, P <0.01 vs model group).

[0038] Figure 20 The effect of T19 on the expression of insulin signaling pathway proteins in STZ-induced diabetic mice (Model 1) / DB / DB mice (Model 2).

[0039] Figure 21 -A represents the effect of T19 on phosphorylation of insulin signaling molecules in diabetic mice; Figure 21 -C and D are key proteins of the AMPK pathway in the liver studied by Western blotting; Figure 21 -B, E represent the quantitative expression of protein ( P <0.001 vs Ctrl group; # P <0.05, ## P <0.01, ### P <0.001, vs model group), where A represents the normal group; B represents the diabetes model group; C represents T19 (30 mg / kg); D represents T19 (60 mg / kg); E represents Met (30 mg / kg).

[0040] Figure 22 -A represents PCoA, NMDS, and PCA analysis of gut microbiota based on weighted UniFrac; Figure 22-B represents a phylum-level bacterial taxonomic profile analysis of gut bacteria from different groups of mice; Figure 22 -C represents the ratio of hard-walled bacteria to Bacteroides; Figure 22 -D represents a heatmap analysis at the gut microbiota genus level; Figure 22 -E represents the LDA score derived from the LEfSe analysis, indicating an LDA score greater than 2 (the length of the bar represents the LDA score). Figure 22 -F is a Cladogram generated from LEfSe analysis, showing the relationships between taxa (the horizontal line represents the order from the inside to the outside: phylum, class, order, family, genus). Detailed Implementation

[0041] The following non-limiting embodiments are intended to enable those skilled in the art to more fully understand the invention, but do not limit the invention in any way.

[0042] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0043] This invention discloses the application of a rare ginseng ingredient T19 in the preparation of products for the prevention and / or treatment of disorders of glucose and lipid metabolism, wherein the products are food, health products, or pharmaceuticals. Specifically, the use of the claimed rare ginseng ingredient T19 in the preparation of pharmaceuticals for the prevention and / or treatment of disorders of glucose and lipid metabolism includes, but is not limited to, administering an effective amount of the claimed rare ginseng ingredient T19 to patients to prepare pharmaceuticals for the prevention and / or treatment of diseases caused by disorders of glucose and lipid metabolism, alleviating the symptoms of diseases caused by disorders of glucose and lipid metabolism, or delaying the development or onset of diseases caused by disorders of glucose and lipid metabolism. The claimed rare ginseng ingredient T19 in the preparation of food and health products for the prevention and / or treatment of disorders of glucose and lipid metabolism includes, but is not limited to, products suitable for daily consumption by specific populations, having the function of regulating the body's glucose and lipid metabolism, preventing disorders of glucose and lipid metabolism, and not causing any acute, subacute, or chronic harm to the human body.

[0044] In one specific embodiment, the product can regulate blood glucose levels, improve lipid metabolism, and alleviate insulin resistance.

[0045] In one specific embodiment, the application of ginseng rare ingredient T19 in the preparation of products for preventing glucose and lipid metabolism disorders refers to the use of the product to prevent or inhibit the occurrence of glucose and lipid metabolism disorders in the presence of factors that may cause abnormal glucose and lipid metabolism (such as high-sugar and high-fat diets, lack of exercise, etc.).

[0046] In one specific embodiment, the application of the rare ginseng ingredient T19 in the preparation of products for treating disorders of glucose and lipid metabolism refers to reducing abnormal blood glucose and blood lipid levels, improving insulin resistance, and alleviating tissue damage caused by disorders of glucose and lipid metabolism.

[0047] In one specific embodiment, glucose and lipid metabolism disorders include hyperglycemia, hyperlipidemia, insulin resistance, and related metabolic abnormalities. Specifically, hyperglycemia is characterized by elevated fasting blood glucose, postprandial blood glucose fluctuations, and elevated glycated hemoglobin; hyperlipidemia is characterized by elevated total cholesterol, triglycerides, and low-density lipoprotein cholesterol, and decreased high-density lipoprotein cholesterol; insulin resistance is characterized by decreased sensitivity to insulin, requiring the secretion of more insulin to maintain normal blood glucose levels.

[0048] In one specific embodiment, the concentration of the rare ginseng component T19 in the product is 20 mg / kg body weight to 60 mg / kg body weight. Preferably, the concentration of the rare ginseng component T19 in the product is 30 mg / kg body weight to 60 mg / kg body weight.

[0049] In one specific embodiment, the food, health product, or pharmaceutical is a composition comprising ginseng rare ingredient T19 as the active ingredient and excipients acceptable to food, health product, or pharmaceutical. Specifically, the excipients acceptable to food, health product, or pharmaceutical are well known in the art, and those skilled in the art can determine that they meet clinical standards.

[0050] In one specific embodiment, when the rare ginseng ingredient T19 is used as an active ingredient in the preparation of foods, health products, or medicines for the prevention and / or treatment of disorders of glucose and lipid metabolism, it can be prepared into one or both of oral and injectable dosage forms.

[0051] In one specific embodiment, oral and injectable dosage forms can be formulated into any conventional dosage form.

[0052] In one specific embodiment, the oral dosage form includes one or more of tablets, capsules, granules, syrups and emulsions, thus making it suitable for various application scenarios and having a good market prospect.

[0053] In one specific embodiment, the injection methods for injectable dosage forms include subcutaneous injection, intramuscular injection, and intravenous injection.

[0054] In one specific embodiment, the rare ginseng component T19 is a product obtained by hydrolyzing and recrystallizing the total saponins obtained by separating ginseng or American ginseng.

[0055] Example 1 This embodiment analyzes the effects of T19 on body weight, blood glucose, glucose tolerance, and blood lipids in STZ-induced diabetic mice (in vivo experiment). The experimental method is as follows: A diabetic mouse model was established using multiple small-dose injections of STZ: 90 four-week-old C57 / BL6 mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. All animals were allowed free access to food and water at 20℃~24℃ and were housed in cages with a 12h:12h light / dark cycle and 40%~60% humidity, with daily water and bedding changes. After 1 week of acclimatization, mice were intraperitoneally injected with streptozotocin (STZ, dissolved in freshly prepared 0.1 μM citrate buffer) at a dose of 50 mg / kg body weight three times, with each injection spaced two days apart, to induce a diabetic mouse model. After 16 hours of fasting, a drop of venous blood was collected from the tail of each mouse to measure its blood glucose level. Mice with blood glucose levels greater than 16.7 mmol / L and exhibiting polydipsia, polyphagia, and polyuria were selected as ideal experimental diabetic models. Animal grouping and administration experiments: The control group (Ctrl) consisted of 10 well-fed 4-week-old C57 / BL mice fed a normal diet; the model group (STZ) consisted of 10 STZ-induced diabetic mice fed a normal diet; the T19 low-dose group (20 mg / kg body weight) consisted of 10 STZ-induced diabetic mice administered T19 (20 mg / kg body weight) by gavage every other day; the T19 high-dose group (40 mg / kg body weight) consisted of 10 STZ-induced diabetic mice administered T19 (40 mg / kg body weight) by gavage every other day; and the metformin group (Met) consisted of 50 mg / kg body weight. The administration period was 4 weeks. Following the animal grouping and administration methods described above, mouse body weight was recorded weekly after one week of administration until the end of the administration period, and significant differences between groups were compared. Blood glucose levels were also recorded after 16 hours of fasting weekly after administration until the end of the administration period, and significant differences between groups were compared. At the end of the dosing cycle, mice were fasted for 12 h and orally administered 2.0 g / kg glucose. Glucose levels in blood samples collected from the tail vein were measured at 0, 30, 60, 90, and 120 min. The area under the curve (AUC) was used to quantify the OGTT results, and significant differences between groups were compared. After the dosing cycle, 1 mL of blood was collected from the retroocular venous plexus of each group of mice. After standing at room temperature for 30 min, the plasma was separated at 4°C, 4000 rpm / min × 15 min. The separated plasma was placed in a 1.5 mL cryovial, sealed, and stored at -80°C for later use. Insulin, triglyceride, and total cholesterol levels in mouse serum were tested according to the instructions on the insulin, TG, and TC kits. Each sample was tested three times to eliminate systematic errors, and the data were analyzed to compare significant differences between groups.The experimental results are as follows. Figure 1-4 As shown.

[0056] like Figure 1 As shown, in the STZ model (Model 1), high dose of T19 (40 mg / kg body weight) significantly reduced the body weight of mice in the fourth week after administration, with no significant changes at other times.

[0057] like Figure 2 As shown, in the STZ model (Model 1), blood glucose levels in the model group increased significantly in the fourth week. In the treatment group, only high-dose (40 mg / kg body weight) T19 was administered again in the fourth week, which significantly reduced blood glucose levels in mice.

[0058] like Figure 3 As shown, in the glucose tolerance test of the STZ model (Model 1), the administration of T19 inhibited blood glucose levels, but the effect was not significant, which may be related to the modeling mechanism of STZ.

[0059] like Figure 4 As shown, in the STZ model (Model 1), blood lipids and total cholesterol increased significantly due to STZ injection. However, the administration of two doses of T19 significantly reduced blood lipids and total cholesterol in STZ-type diabetic mice, with the higher dose showing a more pronounced effect. This confirms that T19 has a good lipid-lowering function and is more effective than metformin.

[0060] Therefore, according to Figure 1-4 Experimental results showed that T19 could improve weight gain, blood glucose elevation, blood lipid and total cholesterol elevation in STZ-induced diabetic mice.

[0061] Example 2 This embodiment analyzes the effects of T19 on body weight, blood glucose, glucose tolerance, and blood lipids in DB / DB mice (in vivo experiment). The experimental methods are as follows: Gene-knockout DB / DB spontaneous type 2 diabetes model mice: 15 nine-week-old C57BL / KsJ-DB / DB mice were purchased from Nanjing Junke Biotechnology Co., Ltd.; 5 nine-week-old C57BL / KsJ mice were purchased from Nanjing Junke Biotechnology Co., Ltd. All DB / DB mice had blood glucose levels above 15 mmol / L, were nine weeks old, and were mature type 2 diabetes models. All animals were allowed free access to food and water at 20℃~24℃, and were housed in cages with a 12h:12h light / dark cycle and a humidity of 40%~60%, with daily water and bedding changes. Based on uniform blood glucose levels, the control group (Ctrl) consisted of 5 normally fed and well-treated C57BL / KsJ mice; the model group consisted of 5 nine-week-old C57BL / KsJ-DB / DB mice, normally fed; the T19 group consisted of 5 nine-week-old C57BL / KsJ-DB / DB mice, administered T19 (40 mg / kg body weight) by gavage every other day; and the rosiglitazone group consisted of 5 nine-week-old C57BL / KsJ-DB / DB mice, administered rosiglitazone (4 mg / kg body weight) by gavage every other day. The administration period was 5 weeks. Following the animal grouping and administration methods described above, after one week of administration, the mouse body weight was recorded weekly until the end of the administration period, and significant differences between groups were compared. Blood glucose levels were also recorded weekly after 16 hours of fasting until the end of the administration period, and significant differences between groups were compared. At the end of the administration period, mice were fasted for 12 hours and orally administered 2.0 g / kg glucose. Glucose levels in blood samples collected from the tail vein were measured at 0, 30, 60, 90, and 120 min. The area under the curve (AUC) was used to quantify the OGTT results, and significant differences between groups were compared. After the dosing cycle, 1 mL of blood was collected from the retroocular venous plexus of each group of mice. After standing at room temperature for 30 min, the plasma was separated at 4°C, 4000 rpm / min × 15 min. The separated plasma was placed in 1.5 mL cryovials, sealed, and stored at -80°C for later use. Insulin, triglyceride, and total cholesterol levels in mouse serum were tested according to the instructions on the insulin, TG, and TC kits. Each sample was tested three times to eliminate systematic errors. Data were analyzed to compare significant differences between groups. Experimental results are as follows: Figure 5-8 As shown.

[0062] like Figure 5 As shown, in the DB / DB model (Model 2), based on the results of the STZ model (Model 1), a high dose of T19 was administered via gavage every other day for five weeks. The results showed that T19 did not affect body weight, possibly due to high food intake and low physical activity.

[0063] like Figure 6 As shown, in the DB / DB model (Model 2), the blood glucose level of mice in the model group remained high. T19 could significantly reduce the blood glucose level of mice and had good stability, reaching a level similar to that of the positive control drug rosiglitazone.

[0064] like Figure 7 As shown, in the glucose tolerance test of DB / DB mice (model 2), T19 administration significantly inhibited the rise in blood glucose and enhanced glucose tolerance, with significant effects at 30 and 60 min. AUC more directly showed that T19 significantly enhanced the glucose tolerance of mice.

[0065] like Figure 8 As shown, in DB / DB mice (model 2), the total cholesterol level increased significantly. Furthermore, the experiment found that administration of T19 could reduce the total cholesterol level in DB / DB mice, restoring it to a state that was not significantly different from the normal group. The effect was no different from that of the positive control drug. T19 did not significantly change serum triglycerides in the DB / DB mouse model.

[0066] Therefore, according to Figure 5-8 Experimental results showed that T19 could improve elevated blood glucose, glucose tolerance, and total cholesterol in DB / DB diabetic mice.

[0067] Example 3 This embodiment analyzes the effects of T19 on body weight, blood glucose, glucose tolerance, and blood lipids in a type 2 diabetic mouse model established by a combination of high-fat, high-glucose diet and repeated low-dose STZ injections (in vivo experiment). The experimental method is as follows: Ninety four-week-old C57 / BL6 mice were purchased from Liaoning Changsheng Biotechnology Co., Ltd. All animals were allowed free access to food and water at 20℃~24℃ and were housed in cages with a 12h:12h light / dark cycle and 40%~60% humidity. Water and bedding were changed daily. After one week of acclimatization, 10 mice were randomly selected as the normal group and fed a normal diet, while the other mice were used as the experimental group and fed a high-fat and high-glucose diet (basal food 49.5%, sucrose 25%, lard 15%, egg yolk powder 5%, milk powder 5%, and sodium cholate 0.5%). After six weeks of feeding, the diabetic mice were fasted for 12 hours and then injected intraperitoneally with streptozotocin (STZ, 50 mg / kg, dissolved in freshly prepared 0.1 μM citrate buffer) for three consecutive days. After fasting for 16 hours, a drop of venous blood was collected from the tail of mice to measure their blood glucose levels. Mice with blood glucose levels greater than 16.7 mmol / L and exhibiting polydipsia, polyphagia, and polyuria were selected as ideal experimental diabetes models. Throughout the experiment, diabetic mice were continuously fed a high-fat, high-sugar diet. Animal grouping and drug administration were as follows: the control group consisted of 10 well-fed 4-week-old C57 / BL mice fed a normal diet; the model group consisted of 10 mice fed a high-fat, high-sugar (HFD) diet and STZ-induced hyperglycemia, fed a high-fat diet; based on previous experimental results, the T19 low-dose group (30 mg / kg body weight) consisted of 10 HFD / STZ-induced hyperglycemic mice administered daily by gavage; the T19 high-dose group (60 mg / kg body weight) consisted of 10 HFD / STZ-induced hyperglycemic mice administered daily by gavage; and the metformin group (Met) was administered metformin (30 mg / kg body weight). The drug administration period was 6 weeks. Based on the animal grouping and administration methods mentioned above, after one week of administration, the mouse body weight was recorded weekly until the end of the administration cycle, and significant differences between groups were compared. Blood glucose levels were also recorded after 16 hours of fasting following weekly administration, and significant differences between groups were compared until the end of the administration cycle. At the end of the administration cycle, mice were fasted for 12 hours and orally administered 2.0 g / kg glucose. Glucose levels were measured in blood samples collected from the tail vein at 0, 30, 60, 90, and 120 minutes. Insulin tolerance was measured using a similar method. After overnight fasting, the blood glucose response to intraperitoneal injection of 0.75 U / kg insulin was determined. Blood samples collected from the tail vein at 0, 30, 60, 90, and 120 minutes after insulin treatment were measured for glucose. The area under the curve (AUC) was used to quantify OGTT and ITT results. Significant differences between groups were compared.Significant differences between groups were compared. After the dosing cycle, 1 mL of blood was collected from the posterior venous plexus of each mouse group. After standing at room temperature for 30 min, the plasma was separated at 4℃, 4000 rpm / min × 15 min. The separated plasma was placed in a 1.5 mL cryovial, sealed, and stored at -80℃ for later use. The levels of insulin, triglycerides, and total cholesterol in mouse serum were tested according to the instructions on the insulin, TG, and TC kits. Each sample was tested three times to eliminate systematic errors. The data were analyzed to compare significant differences between groups. The experimental results are as follows. Figure 9-13 As shown.

[0068] like Figure 9 As shown, in the model established by multiple low-dose STZ injections (Model 3) combined with a high-fat, high-sugar diet, mice fed a high-fat, high-sugar diet (HFD) gradually gained weight compared to the normal group during the first 6 weeks of the feeding period. Subsequently, mice fed an HFD diet were given low-dose STZ injections to induce a type 2 diabetic mouse model. Throughout the study, the normal group mice continued to gain weight, while the model group lost weight due to diabetes. After 6 weeks of drug treatment, the metformin group, and the low-dose and high-dose T19 groups all had significantly higher body weights than the diabetic model group. These results demonstrate that T19 can reduce weight loss in diabetic mice.

[0069] like Figure 10 As shown, in the model established by multiple low-dose injections of STZ in a high-fat, high-glucose diet (Model 3), fasting blood glucose levels in normal mice remained at normal levels throughout the study, while blood glucose levels in model mice were all above 11.3 mmol / L. After 6 weeks of treatment with low-dose T19, high-dose T19, and Met, FBG decreased by 39.2%, 29.6%, and 33.6%, respectively.

[0070] like Figure 11 As shown in the glucose tolerance test results of Model 3, blood glucose levels in all groups peaked at approximately 30 minutes. The diabetic mice exhibited significant glucose intolerance. Compared to the diabetic model group, the T19 group showed a significant decrease in blood glucose at 120 minutes, while the area under the curve (AUC-OGTT) of the T19 (30 mg / kg) and T19 (60 mg / kg) groups decreased by 33.6% and 50.2%, respectively.

[0071] like Figure 12As shown, after insulin injection, the diabetes model groups induced by HFD and low-dose STZ exhibited higher blood glucose levels and area under the curve (AUC-ITT) than the normal control group. In particular, the blood glucose reduction rates in both the high- and low-dose T19 treatment groups were faster than in the diabetes control group. OGTT and ITT are important indicators of the body's response to acute hyperglycemia and are commonly used to assess glucose metabolism. This result indicates that T19 can regulate glucose absorption and metabolism.

[0072] like Figure 13 As shown, in high-fat, high-glucose mice induced by STZ (Model 3), compared with the normal group, the serum TG, TC, and LDL levels of diabetic mice were significantly increased, while the HDL level was significantly decreased, indicating significant lipid abnormalities in diabetic mice. However, treatment with T19 (60 mg / kg) for six weeks significantly reduced TG, TC, and LDL-C levels (31.6%, 24.3%, 17.3%) and increased HDL-C levels (30.9%).

[0073] Therefore, according to Figure 9-13 Experimental results showed that T19 could improve weight loss, blood glucose elevation, glucose tolerance, glucose absorption and metabolism, and dyslipidemia in diabetic mice induced by high-fat, high-glucose combined with STZ.

[0074] Example 4 This embodiment examines the protective effect of T19 on the liver of diabetic mice through pathological examination (in vitro experiment). The pathological testing was provided by Shanghai Unimicron Biotechnology Co., Ltd. The experimental results are as follows: Figure 14-15 As shown.

[0075] like Figure 14 As shown, liver sections of model 2 mice showed obvious enlargement of fat granules and changes in cell nucleus shape. Both T19 and rosiglitazone could improve this condition, and T19 was more effective than the positive control drug.

[0076] like Figure 15 As shown in Figure A, the liver tissue of diabetic mice induced by a high-fat, high-sugar diet in Model 3 exhibited significant destructive changes, including nuclear enlargement, cytoplasmic or vacuolar swelling, bullous steatosis, and some ballooning changes. However, in the groups treated with T19 or Met, vacuolar degeneration and hepatocyte steatosis were significantly reduced, indicating that the liver was protected from damage to some extent.

[0077] like Figure 15 As shown in Figure B, the area of ​​glycogen in the liver stained with PAS was significantly reduced in the T19 or Met groups compared to the diabetic model group. Therefore, T19 can protect the liver from damage by regulating lipid and glycogen deposition in diabetic mice, with effects comparable to metformin.

[0078] Therefore, according to Figure 14-15 Experimental results show that T19 has a protective effect on the liver of diabetic mice.

[0079] Example 5 This example analyzes the effect of T19 on liver inflammation in diabetic mice (in vitro experiment). 50 mg of liver tissue, after being ground in liquid nitrogen, was placed in an EP tube containing 1 mL of Trizol-free RNase and lysed on ice for 1 h. 200 μL of pre-chilled chloroform was added, and the tube was incubated at room temperature for 5 min, vortexed for 1 min, and centrifuged at 4°C × 12000 rpm / min for 15 min. Approximately 600 μL of the intermediate pink aqueous phase was placed in a new RNase-free EP tube. 600 μL of pre-chilled isopropanol was added, the tube was vortexed for 1 min, and incubated at room temperature for 10 min. The tube was centrifuged at 4°C × 12000 rpm / min for 30 min, and the supernatant was carefully aspirated, leaving the precipitate. The precipitate was resuspended in 90% ethanol prepared with RNase-free water and centrifuged at 4°C × 12000 rpm / min for 15 min. The supernatant was carefully aspirated, and the tube opening was placed in a sealed, non-ventilated space to allow the ethanol to evaporate. Total RNA from skeletal muscle was obtained by dissolving the RNA precipitate in 100 μL of RNase-free water, mixing well, and storing at -80℃. 1 μL of RNA sample was taken and its concentration was determined using a micro-Cary UV-Vis spectrophotometer. Total RNA was reverse transcribed according to the cDNA first-strand synthesis kit instructions. RNA template, Primer Mix, dNTPMix, RT Buffer, SuperRT, and RNase-Free Water were dissolved and placed on ice. The reaction mixture was prepared according to the instructions, with a total volume of 20 μL. The mixture was vortexed to mix, briefly centrifuged to collect the solution from the tube wall to the bottom. The mixture was incubated at 42℃ for 30-50 minutes and then at 85℃ for 5 minutes. After the reaction, the mixture was briefly centrifuged and cooled on ice. It was stored at -20℃. Primer design and Real-Time PCR were performed using Primer 5.0 software. RNA from mouse liver tissue in each group was tested using the above method. The experimental results were collected and analyzed after the experiment, and significant differences between each group were compared. The experimental results are as follows. Figure 16-17 As shown.

[0080] like Figure 16 As shown, in the STZ mouse model, both doses of T19 significantly reduced the transcriptional levels of IL-1, IL-6, and COX-2, and the effect was significantly better than that of metformin. However, neither metformin nor T19 had any effect on TNF-α.

[0081] like Figure 17As shown, in the DB / DB mouse model, T19 significantly reduced the transcriptional levels of IL-1, IL-6, and TNF-α.

[0082] Therefore, according to Figure 16-17 Experimental results showed that T19 significantly inhibited the transcription of liver inflammatory factors, with effects comparable to positive control drugs.

[0083] Example 6 This example analyzes the effects of T19 on the transcriptional levels of GLUT4 and AMPK in the skeletal muscle of diabetic mice (in vitro experiment). 50 mg of liver tissue, after being ground in liquid nitrogen, was placed in an EP tube containing 1 mL of Trizol-free RNase and lysed on ice for 1 h. 200 μL of pre-chilled chloroform was added, and the tube was incubated at room temperature for 5 min, vortexed for 1 min, and centrifuged at 4°C × 12000 rpm / min for 15 min. Approximately 600 μL of the intermediate pink aqueous phase was transferred to a new RNase-free EP tube. 600 μL of pre-chilled isopropanol was added, the tube was vortexed for 1 min, and incubated at room temperature for 10 min. The tube was centrifuged at 4°C × 12000 rpm / min for 30 min, and the supernatant was carefully aspirated, leaving the precipitate. The precipitate was resuspended in 90% ethanol prepared with RNase-free water and centrifuged at 4°C × 12000 rpm / min for 15 min. The supernatant was carefully aspirated, and the tube opening was placed in a sealed, non-ventilated space to allow the ethanol to evaporate. Total RNA from skeletal muscle was obtained by dissolving the RNA precipitate in 100 μL of RNase-free water, mixing well, and storing at -80℃. 1 μL of RNA sample was taken and its concentration was determined using a micro-Cary UV-Vis spectrophotometer. The total RNA was reverse transcribed according to the cDNA first-strand synthesis kit instructions. RNA template, Primer Mix, dNTP Mix, RT Buffer, SuperRT, and RNase-Free Water were dissolved and kept on ice. The reaction mixture was prepared according to the instructions, with a total volume of 20 μL. The mixture was vortexed to mix, briefly centrifuged to collect the solution from the tube wall to the bottom. The mixture was incubated at 42℃ for 30-50 minutes and then at 85℃ for 5 minutes. After the reaction, the mixture was briefly centrifuged and cooled on ice. It was stored at -20℃. Primer design and Real-Time PCR were performed using Primer 5.0 software. RNA in mouse skeletal muscle was tested using the above method in each group. The experimental results were collected and analyzed after the experiment, and significant differences between each group were compared. The experimental results are as follows. Figure 18-19 As shown.

[0084] like Figure 18As shown, in STZ-induced diabetic mice (Model 1), AMPK in skeletal muscle was significantly lower than that in normal mice, but significantly higher than that in the model group after treatment with high concentrations of T19, which was the same as that in normal mice. Although there was no significant change in GLUT4 in skeletal muscle of the model group mice, it was significantly higher than that in the model group after treatment with high concentrations of T19.

[0085] like Figure 19 As shown, in DB / DB diabetic mice (model 2), AMPK and GLUT4 levels were significantly lower in the model group than in the normal group, and both levels increased after T19 treatment, but the effect was not as significant as in the STZ model.

[0086] Therefore, according to Figure 18-19 Experimental results showed that T19 treatment could upregulate the transcriptional levels of GLUT4 and AMPK in the skeletal muscle of diabetic mice.

[0087] Example 7 This embodiment analyzes the effect of T19 on the expression of insulin signaling pathway proteins in diabetic mice (in vitro experiment). Skeletal muscle / liver samples were removed from a -80℃ freezer and placed on ice. They were then ground into powder using liquid nitrogen, and 20 mg of the ground tissue was weighed as soon as possible. 200 μL of protein lysis buffer (RIPA lysis buffer: PMSF = 99:1) was added to the tissue sample, and lysis was performed on ice for 3 h. The samples were then centrifuged at 4℃ × 12000 rpm / min for 15 min using a low-temperature high-speed centrifuge, and the supernatant was transferred to a new EP tube. 5 μL of protein sample was taken and its concentration was determined using a BCA protein concentration kit. The remaining protein samples were mixed with 40 μL of 5× Loading Buffer and incubated at 95℃ for 5 min to denature the proteins. The denatured protein samples were labeled and stored at -80℃ for Western blotting experiments. A 1 mm gel was washed with ddH2O onto a glass plate, and the gel-contact side was tilted downwards and placed on a clean paper towel to air dry. The electrophoresis apparatus was then assembled. Prepare 5 mL of 10% separating gel according to the instructions and pour it into the center of the assembled gel plate. Seal the gel with 200 μL of isopropanol and incubate at 25°C for 30 min to allow the separating gel to solidify. Prepare 3 mL of stacking gel, insert a comb, and remove the comb after 30 min. Note that when removing the comb, add water while removing it to prevent air from breaking the comb holes. Take out the denatured protein sample and aspirate 20 mg according to the concentration into a new EP tube. Incubate the tube at 40°C for 10 min before loading. Assemble the gel plate and place it in the electrophoresis tank. Add electrophoresis buffer and load the sample for electrophoresis. After electrophoresis, transfer the membrane using a wet transfer method. After the membrane is transferred, place it in a shaker with a pre-prepared 5% BSA solution and block at room temperature for 3 h before incubating with antibody. Each antibody was diluted 1:1000 with 5% BSA solution, and the β-tublin antibody was diluted 1:5000. After incubation for 3 h, the membrane was washed with TBST three times for 30 min each time, then incubated with secondary antibody at a 1:3000 solution for 2.5 h before exposure. Proteins extracted from mice in each group were tested using the above method, and the results were compared to identify significant differences between groups. Experimental results are as follows: Figure 20-21 As shown.

[0088] like Figure 20 As shown, compared with the normal group, the expression of p-IR in diabetic mice of models 1 and 2 was significantly reduced, and the expression of downstream p-INS-1 and p-AKT was also significantly decreased. However, after administration of T19, the expression of phosphorylated proteins of IR, INS-1, and AKT was increased. This indicates that T19 can effectively activate the insulin signaling pathway, enhance the binding of insulin and insulin-like growth factor to IR, promote its phosphorylation, promote GLUT4 transport, and enhance glucose metabolism. However, no significant changes were observed in GSK3β in models 1 and 2. This suggests that T19 does not change the total expression level of GSK3β, and its effect on phosphorylation requires further investigation.

[0089] like Figure 21 As shown in Figures A and B, in diabetic model 3, the expression levels of p-PI3K, p-AKT, and p-IRS in the liver of the insulin signaling pathway were significantly decreased, while the expression of phosphorylated GSK3β protein was significantly increased. However, T19, to some extent, increased the expression of p-IRS, p-IR, and p-AKT in the liver of diabetic model mice and attenuated the expression of p-GSK3β in the liver. In this study, compared with the diabetic model group, T19 upregulated the protein expression of the key glucose transporter GLUT4.

[0090] like Figure 21 As shown in C, D, and E, the protein expression of AMPK, a key target of glucose and lipid metabolism, was studied in HFD / STZ-induced diabetic mice (model 3). The expression of major proteins in the AMPK pathway, including AMPK, p-AMPK, ACC, and p-ACC, was tested. Western blot analysis was used to analyze the activation effect of T19 on AMPK and ACC. The results showed that the phosphorylation levels of AMPK and ACC were decreased in the diabetic model group, while T19 significantly increased their phosphorylation expression.

[0091] Therefore, according to Figure 20-21 Experimental results show that T19 can effectively activate the insulin signaling pathway, enhance the binding of insulin and insulin-like growth factor to IR, promote its phosphorylation, promote GLUT4 transport, enhance glucose metabolism, upregulate the protein expression of the key glucose transporter GLUT4, and increase the phosphorylation expression of AMPK and ACC.

[0092] Example 8 This embodiment analyzes the effects of T19 on the gut microbiota (in vitro experiment). The role of gut microbiota in human physiological functions has been well-established. Diabetes can significantly affect the diversity of gut bacteria. To test whether T19 can alter the distribution of gut microbiota, type 2 diabetic mice induced by HFD / STZ were tested. T19 was administered for 6 weeks, and fresh feces were collected after the experiment for differential analysis between groups. The gut microbiota detection and analysis were provided by Shanghai Panoson Biotechnology Co., Ltd. The experimental results are as follows: Figure 22 As shown.

[0093] like Figure 22 As shown in -A, the impact of T19 treatment on the gut microbiota was assessed using high-throughput sequencing of the bacterial 16S rDNA V3-V4 region. Figure 22As shown in -A, the degree of difference in gut microbiota distribution among the groups (DM, DM + T19, and NC groups) was assessed based on weighted UniFrac principal coordinate analysis (PCoA), non-metric dimension score (NMDS), and principal component analysis (PCA). The results of PCoA, NMDS, and PCA showed significant clustering of microbial composition in each group, indicating that administration of T19 had a significant impact on the gut microbiota profile of mice.

[0094] like Figure 22 As shown in B and C, in the fecal microbial community, the phylum Firmicutes ( Firmicutes ) and Bacteroidetes ( Bacteroidetes Firmicutes (FCL) is the most abundant phylum. Compared with the normal group, the diabetic model group showed a significant increase in the abundance of Firmicutes and a significant decrease in the abundance of Bacteroidetes. A significant change in the ratio of Firmicutes to Bacteroidetes was observed in the diabetic model mice compared with normal mice. T19 treatment led to a significant decrease in the ratio of Firmicutes to Bacteroidetes, indicating that T19 can remodel the structure of the gut microbiota.

[0095] like Figure 22 As shown in Figure -D, the heatmap with the average abundance of the top 50 genera further indicates that T19 treatment altered the gut microbiota composition of HFD / STZ-induced diabetic mice. Based on heatmap analysis, the effect of bacterial genera on the distribution of gut microbiota in the three experimental groups was assessed. In the T19 group, […]. Coprobacillus , Streptococcus , Lactobacillus , Ruminococcus , Anaerotruncus , Roseburia and Coprococcus The rich variety of bacteria is positively correlated with glucose and lipid metabolism.

[0096] like Figure 22 As shown in Figure -E, the abundance of gut microbiota was assessed using the linear discriminant analysis effect size (LEfSe) method, and significant differences were found among the three groups. LEfSe analysis with a log LDA score threshold of 2 was performed to identify specific bacteria associated with T19 administration, determining specific bacterial taxa at the phylum to genus level in each group.

[0097] like Figure 22 As shown in -F, the characteristics of the normal control group are: Anaerofustis , Rikenellaceae , Coriobacteriales , Bacteroidia , Bacteroidetes The proportion was relatively high, while in the diabetes model group, the gut microbiota was rich in... Oscillospira , Ruminococcaceae , Desulfovibrionales , Clostridia , GemmatimonadetesIt is worth noting that in the DM-T19 group, the characteristic of T19 administration is also that... Oligella , Lachnospiraceae , Pseudomonadales , Actinobacteria , Actinobacteria The changes.

[0098] Therefore, according to Figure 22 Experimental results showed that T19 could significantly alter the distribution of gut microbiota in type 2 diabetic mice induced by HFD and STZ.

[0099] In summary, this invention, through studies on three diabetic mouse models using T19, revealed that T19 regulates glucose and lipid metabolism. The potential mechanism of this effect may be related to the activation of the AMPK / PI3K signaling pathway, improving insulin resistance, and positively influencing gut microbiota regulation, thereby modulating blood glucose and lipid levels, indicating that T19 has the potential to treat type 2 diabetes.

Claims

1. Application of rare ginseng component T19 in the preparation of products for the prevention and / or treatment of disorders of glucose and lipid metabolism.

2. The application according to claim 1, characterized in that: The effects of the products for preventing and / or treating disorders of glucose and lipid metabolism include at least one of the following: (a) Improve blood glucose and / or lipid metabolism, including lowering fasting blood glucose, improving glucose tolerance, and lowering serum total cholesterol and / or triglyceride levels; (b) Improve abnormal weight; (c) Protect the liver and improve liver damage, including reducing hepatic lipid accumulation and inhibiting the transcription of hepatic inflammatory factors; (d) Activation of the insulin signaling pathway, including promoting insulin receptor phosphorylation, upregulating the expression or transcription of glucose transporter 4, and / or upregulating the transcription and phosphorylation levels of AMP-activated protein kinase; (e) Regulate the gut microbiota to normalize its structure.

3. The application according to claim 1, characterized in that: The glucose and lipid metabolism disorders include hyperglycemia, hyperlipidemia, insulin resistance, and related metabolic abnormalities.

4. The application according to claim 3, characterized in that: The disorder of glucose and lipid metabolism is diabetes.

5. The application according to claim 1, characterized in that: The rare ginseng component T19 is a product obtained by hydrolyzing and recrystallizing the total saponins obtained by separating ginseng or American ginseng.

6. The application according to claim 1, characterized in that: The concentration of the rare ginseng ingredient T19 in the product is 20~60 mg / kg body weight.

7. The application according to claim 1, characterized in that: The product in question is food, health supplement, or medicine.

8. The application according to claim 7, characterized in that: The product includes the active ingredient ginseng rare ingredient T19 and other excipients acceptable in food, health products or pharmaceuticals.

9. The application according to claim 7, characterized in that: The product is available in oral and injectable dosage forms.

10. The application according to claim 9, characterized in that: The oral dosage form is a tablet, capsule, granule, syrup, or emulsion; the injectable dosage form is administered via subcutaneous injection, intramuscular injection, or intravenous injection.

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