Kunlun baby chrysanthemum polysaccharide with sugar control and lipid-lowering effects and a preparation method thereof

Abstract

The application provides a Kunlun baby chrysanthemum polysaccharide with a blood sugar control and lipid reduction effect and a preparation method thereof, relates to the technical field of biological medicine, and the main chain of the Kunlun baby chrysanthemum polysaccharide is composed of→4)-beta-D-Galp-(1→ and→4)-beta-D-Glcp-(1→ alternately; the side chain of the Kunlun baby chrysanthemum polysaccharide is composed of→5)-alpha-L-Araf-(1→,→3,6)-beta-D-Galp, alpha-L-Araf, beta-D-Glcp / Galp and alpha-D-Rhap. The application solves the technical problem that the prepared Kunlun baby chrysanthemum polysaccharide in the prior art is complex in composition and low in purity, and seriously limits the application of the Kunlun baby chrysanthemum polysaccharide in the field of blood sugar control and lipid reduction, so that the application achieves the effects of reducing blood sugar and reducing blood lipid, is beneficial to reducing the use dosage of the blood sugar reducing drug metformin and enhancing the blood sugar control and lipid reduction effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and in particular to a Kunlun baby chrysanthemum polysaccharide with blood sugar and lipid-lowering effect and a preparation method thereof. BACKGROUND

[0002] Diabetes is a chronic disease characterized by insufficient insulin secretion or impaired insulin utilization, and persistent high blood glucose levels are its main marker. It is often accompanied by various complications such as hyperlipidemia, which poses a threat to the patient's life. In recent years, the incidence and prevalence of diabetes worldwide have increased, posing a major challenge to public health. This disease can cause chronic and progressive damage to tissues and organs, which can lead to functional decline or even failure, and is associated with the risk of acute metabolic disorders. Therefore, high blood glucose and hyperlipidemia need to be treated coordinately.

[0003] In the past few decades, metformin has become an important drug for treating high blood glucose and hyperlipidemia worldwide due to its high cost-effectiveness and significant blood glucose-lowering effect. Although metformin can enhance insulin sensitivity, inhibit gluconeogenesis, promote cellular glucose uptake, and reduce cholesterol to improve high blood glucose and hyperlipidemia symptoms. However, long-term use of metformin can cause gastrointestinal discomfort. Although metformin plays an important role in the treatment of diabetes, it is still necessary to explore new treatment strategies to improve the efficacy of metformin and reduce its dosage, thereby improving the prognosis of patients.

[0004] At present, combination therapy is a method often used in clinical treatment, aiming to maximize the efficacy and minimize the side effects by rationally using multiple drugs. Studies have shown that polysaccharides can not only enhance the immune regulation of drugs, but also enhance the efficacy of anti-diabetic and drugs (such as acarbose, metformin, and sitagliptin). Polysaccharides can regulate glucose metabolism and improve islet cell function, thereby enhancing the therapeutic effect of anti-diabetic drugs, for example, the combination therapy of low-dose metformin and sargassum polysaccharide has a positive effect on high-fat diet and streptozotocin-induced diabetic rats.

[0005] Kunlun baby chrysanthemum can be used as medicine and food, and the polysaccharide in Kunlun baby chrysanthemum has the effect of inhibiting carbohydrate enzymes in vitro and can significantly inhibit the activity of alpha-glucosidase. However, the polysaccharide prepared in the prior art has complex components and low purity, which seriously limits its application in the field of blood glucose and lipid-lowering. Therefore, it is urgent to develop a new Kunlun baby chrysanthemum polysaccharide with blood sugar and lipid-lowering effect and a preparation method thereof to reduce the dosage of metformin and enhance the blood sugar and lipid-lowering effect.

[0006] In view of this, the present application is proposed. SUMMARY

[0007] One of the purposes of the present application is to provide a Kunlun baby chrysanthemum polysaccharide which has good sugar control and lipid-lowering effects and can reduce the dosage of the hypoglycemic drug metformin and enhance the sugar control and lipid-lowering effects.

[0008] The second purpose of the present application is to provide a preparation method of the Kunlun baby chrysanthemum polysaccharide, which is simple, easy to operate and suitable for industrial production.

[0009] The third purpose of the present application is to provide an application of the Kunlun baby chrysanthemum polysaccharide, which is beneficial to enhancing the sugar control and lipid-lowering effects of metformin on diabetic patients and reducing side effects.

[0010] In order to achieve the above-mentioned purposes of the present application, the following technical solutions are adopted:

[0011] In a first aspect, a Kunlun baby chrysanthemum polysaccharide is mainly composed of arabinose, galactose, galacturonic acid, rhamnose and glucose.

[0012] The main chain of the Kunlun baby chrysanthemum polysaccharide is composed of →4)-β-D-Galp-(1→ and →4)-β-D-Glcp-(1→ alternately.

[0013] The side chain of the Kunlun baby chrysanthemum polysaccharide is composed of →5)-α-L-Araf-(1→, →3,6)-β-D-Galp, α-L-Araf, β-D-Glcp / Galp and α-D-Rhap.

[0014] Further, the molar percentage of arabinose, galactose, galacturonic acid, rhamnose and glucose is 26.52:37.96:26.43:5.38:3.71.

[0015] Further, the structure formula of the repeating unit in the Kunlun baby chrysanthemum polysaccharide is as follows:

[0016]

[0017] The uronic acid content of the Kunlun baby chrysanthemum polysaccharide is 26.48±0.11%.

[0018] The Kunlun baby chrysanthemum polysaccharide is an acidic polysaccharide with a molecular weight of 3192 Da.

[0019] In a second aspect, a preparation method of the Kunlun baby chrysanthemum polysaccharide of any one of the above aspects comprises the following steps:

[0020] (a) hydrolyzing the Kunlun baby chrysanthemum under acidic conditions, and then filtering to obtain a filtrate;

[0021] (b) concentrating the filtrate obtained in step (a) and then performing alcohol precipitation, and filtering to obtain a precipitate;

[0022] (c) mixing the precipitate obtained in step (b) with an alkali solution, then filtering, and adjusting the pH of the filtrate to neutral to obtain an extract;

[0023] (d) subjecting the extract obtained in step (c) to ultrafiltration to obtain a filtrate and drying to obtain a crude extract;

[0024] (e) subjecting the crude extract obtained in step (d) to ultrafiltration to obtain a retentate and drying to obtain a crude Kunlun baby's breath polysaccharide;

[0025] (f) subjecting the obtained crude Kunlun baby's breath polysaccharide to treatment by anion exchange chromatography and dialysis to obtain the Kunlun baby's breath polysaccharide.

[0026] Further, in step (a), the method of hydrolysis comprises the following steps:

[0027] After the Kunlun baby's breath is soaked with an acid solution, hydrolysis is performed under the condition of cellulase and coupling of magnetic field and electric field to obtain a hydrolysis solution;

[0028] Preferably, the stirring condition of hydrolysis is stirring at 85°C for 2h;

[0029] Preferably, the cellulase is cellulase with enzyme activity of 800U / g after addition, and the pH value is 5.3;

[0030] Preferably, the magnetic induction intensity of the magnetic field is 0.3T-0.5T;

[0031] Preferably, the alternating voltage frequency of the electric field is 50Hz.

[0032] Further, the filtering mode comprises centrifugation.

[0033] Further, in step (c), the stirring condition of mixing the precipitate with the alkali solution is stirring at 40°C-60°C for 2h-3h.

[0034] Further, in step (d), the molecular cut-off of ultrafiltration is 10000Da;

[0035] Preferably, in step (e), the molecular cut-off of ultrafiltration is 5000Da;

[0036] Preferably, in step (e), the molecular weight of the crude Kunlun baby's breath polysaccharide is 5000Da-10000Da.

[0037] Further, the eluent used in the anion exchange chromatography comprises 0.01mol / L NaOH solution and 3 volumes of 0mol / L, 0.2mol / L, 0.4mol / L, 0.6mol / L, 0.8mol / L and 1.0mol / L NaCl solution.

[0038] In a third aspect, the application provides use of the Kunlun baby's breath polysaccharide in any of the above aspects in the preparation of a blood glucose-lowering and / or blood lipid-lowering drug.

[0039] Compared with the prior art, the application has at least the following beneficial effects:

[0040] The Kunlun baby's breath polysaccharide provided by the application has high purity, clear structural characteristics, and both blood glucose-lowering and blood lipid-lowering effects; the Kunlun baby's breath polysaccharide can better prevent atrophy of islet cells by inhibiting cleaved Caspase-3 and inflammatory factor NLRP-3, can enhance the blood glucose-lowering effect of metformin on diabetic patients and reduce side effects, and is conducive to reducing the dosage of the blood glucose-lowering drug metformin.

[0041] The preparation method of the Kunlun baby's breath polysaccharide provided by the application is simple, easy to operate, and suitable for industrial production; the acid combined with enzyme treatment and the magnetic field and electric field coupling can increase the yield of the Kunlun baby's breath polysaccharide and the release of effective components.

[0042] The application of the Kunlun baby's breath polysaccharide provided by the application is conducive to enhancing the blood glucose-lowering and lipid-lowering effect of metformin on diabetic patients and reducing side effects, and provides a new direction for blood glucose-lowering and blood lipid-lowering drugs or drug adjuvants. BRIEF DESCRIPTION OF DRAWINGS

[0043] In order to more clearly illustrate the specific embodiments of the application or the technical solutions in the prior art, the following will briefly introduce the drawings needed to be used in the specific embodiments or the prior art description. Obviously, the drawings in the following description are some embodiments of the application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0044] Figure 1 The anion exchange chromatography filler column chromatography gradient elution curve of the Kunlun baby's breath polysaccharide provided for the application example 1 is shown in the following figure;

[0045] Figure 2 The IC50 value comparison of the Kunlun baby's breath polysaccharide for whether enzyme treatment and magnetic field and electric field coupling are included in the extraction process of the application example 1 is shown in the following figure;

[0046] Figure 3 The inhibition curve of the purified component obtained in the application test example 1 on alpha-glucosidase is shown in the following figure;

[0047] Figure 4 The determination result figure of monosaccharide composition (A) and molecular weight (B) of the Kunlun baby's breath polysaccharide obtained in the application test example 2 is shown in the following figure;

[0048] Figure 5An infrared spectrum of the Kunlun baby's breath polysaccharide obtained in the test example 3 of the present application;

[0049] Figure 6 An infrared spectrum of the Kunlun baby's breath polysaccharide obtained in the test example 5 of the present application 1 H spectrum (A), 13 C spectrum (B), 1 H- 1 H COSY spectrum (C) and HSQC spectrum (D);

[0050] Figure 7 A schematic diagram of the animal test design scheme obtained in the test example 6 of the present application;

[0051] Figure 8 Effects of the Kunlun baby's breath polysaccharide (denoted as AI) obtained in the test example 6 of the present application on the body weight (A), final body weight (B), pancreas index (C), liver index (D) and kidney index (E) of mice;

[0052] Figure 9 Effects of the Kunlun baby's breath polysaccharide obtained in the test example 6 of the present application on HbA1c (A), insulin content (B), liver glycogen content (C) and muscle glycogen content (D) of diabetic mice;

[0053] Figure 10 Schematic diagrams of H&E staining (A), immunofluorescence staining of mouse pancreas, co-staining of insulin (red) and cleaved Caspase-3 (B) or NLRP-3 (green) (C) of mouse pancreas obtained in the test example 6 of the present application;

[0054] Figure 11 Glycogen staining (A) and glycogen signal (B) of nematodes obtained in the test example 7 of the present application;

[0055] Figure 12 Oil red O staining (A) and oil red O signal (B) of nematodes obtained in the test example 7 of the present application. DETAILED DESCRIPTION

[0056] The technical solutions of the present application will be described clearly and completely below in conjunction with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0057] According to a first aspect of the present application, a Kunlun baby's breath polysaccharide mainly composed of arabinose, galactose, galacturonic acid, rhamnose and glucose is provided;

[0058] The main chain of the Kunlun baby's chrysanthemum polysaccharide is composed of →4)-β-D-Galp-(1→ and →4)-β-D-Glcp-(1→ alternately.

[0059] The side chain of the Kunlun baby's chrysanthemum polysaccharide is composed of →5)-α-L-Araf-(1→, →3,6)-β-D-Galp, α-L-Araf, β-D-Glcp / Galp and α-D-Rhap.

[0060] The Kunlun baby's chrysanthemum polysaccharide has good blood sugar and blood lipid lowering effects, can better prevent islet cell atrophy by inhibiting cleaved Caspase-3 and inflammatory factor NLRP-3, can enhance the blood sugar lowering effect of metformin on diabetic patients and reduce side effects, and is beneficial to reducing the use dose of blood sugar lowering drug metformin.

[0061] In the present application, the molar percentage of arabinose, galactose, galacturonic acid, rhamnose and glucose is 26.52:37.96:26.43:5.38:3.71.

[0062] It should be noted that the structural formula of the repeating unit in the Kunlun baby's chrysanthemum polysaccharide is as follows:

[0063]

[0064]

[0065] In the present application, the uronic acid content of the Kunlun baby's chrysanthemum polysaccharide is 26.48±0.11%, which is an acidic polysaccharide with a molecular weight of 3192 Da.

[0066] In summary, the Kunlun baby's chrysanthemum polysaccharide of the present application is composed of arabinose, galactose, galacturonic acid, rhamnose and glucose in a molar ratio (%) of 26.52:37.96:26.43:5.38:3.71, and is an acidic polysaccharide with a molecular weight of 3192 Da; the results of methylation and nuclear magnetic resonance show that the Kunlun baby's chrysanthemum polysaccharide is mainly composed of residues of →4)-β-D-Galp-(1→, →4)-β-D-Glcp-(1→, →5)-α-L-Araf-(1→, →3,6)-β-D-Galp, α-L-Araf, β-D-Glcp / Galp and α-D-Rhap.

[0067] According to a second aspect of the present application, a preparation method of the Kunlun baby's chrysanthemum polysaccharide of any one of the above is provided, comprising the following steps:

[0068] (a) hydrolyzing the Kunlun baby's chrysanthemum under acidic conditions, and then filtering to obtain a filtrate;

[0069] (b) the filtrate obtained in step (a) is concentrated and then subjected to alcohol precipitation, filtration, to obtain a precipitate;

[0070] (c) the precipitate obtained in step (b) is mixed with an alkali solution, then filtered, and the pH of the filtrate is adjusted to neutral, to obtain an extract;

[0071] (d) the extract obtained in step (c) is subjected to ultrafiltration, to obtain a filtrate and dry the filtrate, to obtain a crude extract;

[0072] (e) the crude extract obtained in step (d) is subjected to ultrafiltration, to obtain a retentate and dry the retentate, to obtain a crude Kunlun baby's chrysanthemum polysaccharide;

[0073] (f) the crude Kunlun baby's chrysanthemum polysaccharide obtained is subjected to treatment by anion exchange chromatography and dialysis, to obtain a Kunlun baby's chrysanthemum polysaccharide.

[0074] The preparation method of the Kunlun baby's chrysanthemum polysaccharide provided by the application is simple, easy to operate, and suitable for industrial production.

[0075] In a preferred embodiment, in step (a), the method of hydrolysis comprises the following steps:

[0076] The Kunlun baby's chrysanthemum is soaked with an acid solution, and then subjected to hydrolysis under the condition of cellulase and coupling of magnetic field and electric field, to obtain a hydrolysis solution.

[0077] In the application, the stirring condition of hydrolysis can be stirring at 85 DEG C for 2h, but is not limited thereto; the cellulase can be cellulase with an enzyme activity of 800 U / g after addition, and the pH value can be 5.3, but is not limited thereto; the magnetic induction intensity of the magnetic field can be 0.3T-0.5T, for example, can be 0.3T, 0.4T, 0.5T, but is not limited thereto; at the same time, the alternating voltage frequency of the electric field can be 50Hz, but is not limited thereto.

[0078] The Kunlun baby's chrysanthemum polysaccharide is prepared by cellulase and coupling of magnetic field and electric field in the application, and under the synergistic cooperation of various process parameters, the yield of the Kunlun baby's chrysanthemum polysaccharide is 5.63±0.37%, the total sugar content is 91.29±1.03%, and the uronic acid content is 26.48±0.11%.

[0079] In a preferred embodiment, the filtration mode includes but is not limited to centrifugation, which is more conducive to the full separation of the product.

[0080] In a preferred embodiment, in step (c), the stirring condition of mixing the precipitate with the alkali solution can be stirring at 40 DEG C-60 DEG C for 2h-3h, which is more conducive to the full mixing of the precipitate and the alkali solution.

[0081] In a preferred embodiment, in step (d), the molecular cutoff of ultrafiltration can be 10,000 Da, but is not limited thereto.

[0082] In a preferred embodiment, in step (e), the molecular cutoff of ultrafiltration can be 5000 Da, but is not limited thereto.

[0083] In this invention, in step (e), the molecular weight of the crude polysaccharide of *Chrysanthemum indicum* can be 5000 Da-10000 Da, for example, 5000 Da, 6000 Da, 7000 Da, 8000 Da, 9000 Da, or 10000 Da, but is not limited thereto.

[0084] In a preferred embodiment, the eluent used in anion exchange chromatography can be composed of a 0.01 mol / L NaOH solution in a volume ratio of 1:3 and NaCl solutions of 0 mol / L, 0.2 mol / L, 0.4 mol / L, 0.6 mol / L, 0.8 mol / L, and 1.0 mol / L, respectively.

[0085] Two components are collected in anion exchange chromatography: AI-1 eluted from 0 mol / L NaCl solution and AI-2 eluted from 0.2 mol / L NaCl solution. AI-2 is purified by dialysis to obtain Kunlun Chrysanthemum polysaccharide.

[0086] According to a third aspect of the present invention, the use of the polysaccharide of Chrysanthemum indicum as described in any of the preceding claims in the preparation of drugs for lowering blood sugar and / or lowering blood lipids is provided.

[0087] The application of Kunlun Chrysanthemum polysaccharide provided by this invention is beneficial to enhance the blood sugar and lipid-lowering effect of metformin on diabetic patients and reduce side effects, providing a new direction for hypoglycemic drugs or drug adjuvants.

[0088] The present invention will be further illustrated below by way of examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or purchased directly from the market.

[0089] Example 1

[0090] A polysaccharide from *Chrysanthemum indicum* was prepared by the following method:

[0091] (a) After soaking the Kunlun Chrysanthemum powder in acid solution, it was stirred for the first time and then hydrolyzed under the synergistic magnetic and electric field coupling of cellulase to obtain hydrolysate;

[0092] The ratio of Kunlun Chrysanthemum powder to acid solution was 1g:10mL; the acid solution used was a 0.1mol / L HCl solution.

[0093] The first stirring condition is stirring at 85℃ for 2h;

[0094] The enzyme activity after adding cellulase is 800U / g, and the pH of the system is adjusted to 5.3 (to ensure enzyme activity);

[0095] Meanwhile, the magnetic field and electric field coupling condition is that the magnetic induction intensity is 0.3T-0.5T at 50℃, the alternating voltage frequency is 50Hz, and the hydrolysis time is 3h-5h;

[0096] The obtained hydrolysis liquid is subjected to first centrifugation, and the precipitate is removed to collect the filtrate;

[0097] The speed of the first centrifugation is 6000rpm-7000rpm, and the time is 20min-30min;

[0098] (b) The filtrate obtained in step (a) is concentrated to obtain a concentrated liquid (solid content is 10wt%-20wt%), and the obtained concentrated liquid is subjected to alcohol precipitation and then second centrifugation to collect the precipitate;

[0099] The alcohol precipitation condition is that 4 times the volume of 95% ethanol is added to the concentrated liquid, and it is placed at 4℃ for 12h;

[0100] The speed of the second centrifugation is 6000rpm-7000rpm, and the time is 10min-20min;

[0101] (c) The precipitate obtained in step (b) is added with alkali solution and subjected to second stirring, third centrifugation, and the pH of the obtained filtrate is adjusted to neutral (7.0-7.5) to obtain an extract;

[0102] The second stirring condition is stirring at 40℃-60℃ for 2h-3h;

[0103] The speed of the third centrifugation is 6000rpm-7000rpm, and the time is 10min-20min;

[0104] (d) The extract obtained in step (c) is subjected to first ultrafiltration (repeated for 2-3 times) to obtain a filtrate and dry to obtain a dry material which is a crude extract;

[0105] The molecular cut-off of the first ultrafiltration is 10000Da, and the filtrate is collected;

[0106] (e) The mixed liquid containing the crude extract is subjected to second ultrafiltration (repeated for 3-5 times) to obtain a cut-off liquid and dry to obtain a dry material which is a Kunlun baby's breath crude polysaccharide;

[0107] The molecular cut-off of the second ultrafiltration is 5000Da, and the cut-off liquid is collected;

[0108] After the retentate is dried, low molecular Kunlun baby's breath crude polysaccharides with a molecular weight of 5000 Da-10000 Da are obtained;

[0109] (f) The solution containing Kunlun baby's breath crude polysaccharides is subjected to anion exchange chromatography and dialysis purification to obtain Kunlun baby's breath polysaccharides;

[0110] It should be noted that two components are collected in the anion exchange chromatography, AI-1 component eluted by 0 mol / L NaCl solution and AI-2 component eluted by 0.2 mol / L NaCl solution, and the AI-2 component is subjected to dialysis purification to obtain Kunlun baby's breath polysaccharides;

[0111] Specifically, the separation and purification method of Kunlun baby's breath crude polysaccharides is as follows:

[0112] (1) Activating the filler and loading the column: When the filler is used for the first time, it needs to be activated with acid and alkali solution. First, 0.1M sodium hydroxide solution, deionized water and 0.01M hydrochloric acid are used for flushing, and then a large amount of deionized water is used for flushing until it is neutral. Then slowly and uniformly pour the chromatography column (3.5cm x 30cm), and then pour the chromatography column (3.5cm x 30cm). After the filler is naturally settled, the excess water on the top is discharged from the lower water outlet to ensure that the filler in the column is not layered and has no air bubbles.

[0113] (2) Sample loading: Prepare 30mL of sample loading (10mg / mL) solution, mix well, centrifuge (8000rpm, 4℃, 10min), and take the supernatant for sample loading. 0, 0.2mol / L, 0.4mol / L, 0.6mol / L, 0.8mol / L and 1.0mol / L NaCl solution are used as eluent for elution, and the elution speed is controlled at 1mL / min. Collect 1 tube every 6mL, and elute 600mL for each solubility.

[0114] The total sugar content of the collected solution is detected by the phenol-sulfuric acid method, and the elution spectrum of Kunlun baby's breath polysaccharides is drawn as follows: Figure 1 It can be seen that two components are collected in the elution process, AI-1 component eluted by 0 mol / L NaCl solution and AI-2 component eluted by 0.2 mol / L NaCl solution.

[0115] According to the main peak distribution of the elution spectrum, the components are combined and collected, and after rotary concentration, the dialysis bag with a molecular weight cutoff of 500 Da is used for dialysis for 36h, and then freeze-dried to obtain the purified components.

[0116] In this embodiment, the effects of enzyme treatment and magnetic field-electric field coupling on the α-glucosidase inhibitory activity of Kunlun baby's breath polysaccharides are explored:

[0117] Take 40 μ concentration 4 mg / mL Kunlun fetal chrysanthemum polysaccharide sample (enzyme treatment, magnetic field and electric field coupling extraction process and no enzyme treatment, magnetic field and electric field coupling) and 30 μL α-glucosidase diluent (37℃, 10 min) for incubation, then add 30 μL substrate p-NPG solution for reaction (37℃, 15 min), finally add 100 μL Na2CO3 solution (0.1M) to terminate the reaction, detect the absorbance value at 405 nm;

[0118] The extraction process without enzyme treatment, magnetic field and electric field coupling is the control group, and the extraction process with enzyme treatment, magnetic field and electric field coupling is the experimental group, and the IC 50 value is calculated.

[0119] The results are shown in Figure 2 The IC 50 value of the control group is 670.02 μg / mL, which is lower than the IC 50 value of the experimental group (533.15 μg / mL), which indicates that increasing enzyme treatment and magnetic field and electric field coupling during extraction can increase the yield of Kunlun fetal chrysanthemum polysaccharide and the release of active ingredients, which is beneficial to enhance the hypoglycemic and hypolipidemic effects.

[0120] Test Example 1

[0121] Determination of α-glucosidase inhibitory activity of Kunlun fetal chrysanthemum polysaccharide:

[0122] The purified components corresponding to the two components (AI-1 component and AI-2 component) in Example 1 (30 μL) were respectively incubated with 30 μL α-glucosidase diluent (37℃, 10 min), then 30 μL substrate p-NPG solution was added for reaction (37℃, 15 min), finally 100 μL Na2CO3 solution (0.1M) was added to terminate the reaction, and the absorbance value was detected at 405 nm, and the inhibition rate was calculated by the following formula, and the results are shown in Figure 3 ;

[0123]

[0124] Among them, A 样品 represents the absorbance value of Kunlun fetal chrysanthemum polysaccharide, α-glucosidase and p-NPG, A 样品空白 represents the absorbance value of Kunlun fetal chrysanthemum polysaccharide and PBS, and A 空白对照 represents the absorbance value of PBS, α-glucosidase and p-NPG.

[0125] From Figure 3As can be seen from Table 1, the purified component AI-1 does not exhibit inhibitory activity on α-glucosidase in the detection concentration range, and the purified component AI-2 does not exhibit inhibitory activity on α-glucosidase in the test concentration range of 0.5 mg / mL to 1 mg / mL; however, as the concentration of the purified component AI-2 increases, the purified component AI-2 exhibits inhibitory activity on α-glucosidase in the test concentration range of 2 mg / mL to 8 mg / mL, and presents a dose-dependent manner, i.e., the purified component AI-2 is selected as the Kunlun baby's breath polysaccharide.

[0126] Test Example 2

[0127] The yield and basic composition of the Kunlun baby's breath polysaccharide are determined as follows:

[0128]

[0129] In the formula, W1 represents the mass of the freeze-dried purified component, and W2 represents the loading mass.

[0130] Total sugar content detection: D-glucose is used as a standard, and the phenol-sulfuric acid method is used to determine the total sugar content of the Kunlun baby's breath polysaccharide.

[0131] Uronic acid content detection: galacturonic acid is used as a standard, and the m-hydroxydiphenyl colorimetric method is used to determine the uronic acid content of the Kunlun baby's breath polysaccharide.

[0132] Monosaccharide composition determination: 5 mg of the Kunlun baby's breath polysaccharide and 2 mL of TFA (3 M) are added to an ampoule for hydrolysis (120°C, 3 h), the hydrolyzed polysaccharide solution is subjected to nitrogen blowing, and 5 mL of water is added for dissolution, dilution and centrifugation (12000 rpm, 5 min); the standard preparation method is the same as above, and the sample is subjected to ion chromatography analysis, as shown in FIG. 1A. Figure 4

[0133] Molecular weight determination: 0.1 mol / L sodium nitrate (containing 0.05% sodium azide) solution is used to prepare a 5 mg / mL CTBPs solution, which is subjected to membrane filtration, and then combined with molecular exclusion gel chromatography, 18-angle laser light scattering and differential detector to analyze the molecular weight of the Kunlun baby's breath polysaccharide, as shown in FIG. 1B. Figure 4

[0134] The determination results are as follows: the yield of the Kunlun baby's breath polysaccharide is 5.63±0.37%, the total sugar content is 91.29±1.03%, and the uronic acid content is 26.48±0.11%; the Kunlun baby's breath polysaccharide is composed of arabinose, galactose, galacturonic acid, rhamnose and glucose in a molar ratio (%) of 26.52, 37.96, 26.43, 5.38 and 3.71, and is an acidic polysaccharide with a molecular weight of 3192 Da.

[0135] Test Example 3​​

[0136] Infrared spectrum determination of Kunlun baby's breath polysaccharide:

[0137] The infrared spectrum analysis of Kunlun baby's breath polysaccharide was carried out by KBr tabletting method, and the scanning range of Fourier infrared spectrometer was 4000cm -1 -400cm -1 . The infrared spectrum chart is shown in Figure 5 .

[0138] As can be seen from Figure 5 , the Kunlun baby's breath polysaccharide has characteristic absorption peaks of polysaccharide in the range of 4000cm -1 -400cm -1 , the strong absorption peak at 3370cm -1 is caused by the stretching vibration of O-H, the absorption peak at 2925cm -1 is caused by the asymmetric stretching vibration of C-H, the absorption peak at 1610cm -1 is the asymmetric stretching vibration of non-ester carbonyl C=O, indicating the presence of uronic acid; the absorption peak at 1417cm -1 is the variable angle vibration of C-H or O-H; the absorption peaks at 1089cm -1 and 1026cm -1 indicate the presence of C-O-C stretching vibration; in addition, the infrared spectrum chart has absorption peaks at 891cm -1 and 634cm -1 , which are attributed to β-glycosidic bond.

[0139] Test Example 4

[0140] Methylation determination of Kunlun baby's breath polysaccharide:

[0141] First, the methylation reaction was carried out, 3mg of Kunlun baby's breath polysaccharide was placed in a glass reaction bottle, 1mL of anhydrous DMSO and anhydrous base solution was quickly added, sealed, ultrasonically dissolved, iodomethane solution was added for reaction (30℃, 60min), then ultrapure water was added to terminate the reaction, and then chloroform was added for extraction. The methylated Kunlun baby's breath polysaccharide was transferred to an ampoule, 1mL of 2M TFA was added, sealed, and placed in an oven for hydrolysis (110℃, 90min). After hydrolysis, TFA was removed, 2mL of distilled water was added, 60mg of sodium borohydride was added, and reduction was carried out for 8h. Acetic acid was added for neutralization, dried by spinning, then 1mL of acetic anhydride was added for acetylation reaction (100℃, 1h), cooled, 3mL of toluene was added, dried by spinning, and the operation was repeated until the residual acetic anhydride was removed. After being dissolved in 3mL of dichloromethane, it was transferred to a separatory funnel, a small amount of distilled water was added and shaken thoroughly, the upper aqueous solution was removed, and the operation was repeated 4 times. Sufficient anhydrous sodium sulfate was added for drying, and then 0.22μm filter membrane was used for filtration. Then GC-MS analysis was carried out, and the working parameters are shown in Table 1.

[0142] Table 1

[0143]

[0144] The main glycosidic bond and connection mode of Kunlun baby's breath polysaccharide are shown in Table 2, and 11 kinds of glycosidic bonds are detected, including non-reducing terminal (T-), which is composed of T-Araf (20.50%), 1, 5-Araf (11.50%), 1, 4-Galp (12.20%), T-Galp (9.80%), 1, 3, 6-Galp (9.60%), 1, 4-Glcp (8.40%), T-Glcp (8.30%), 1, 6-Galp (6.80%), 1, 3-Galp (6.40%), T-Rhap (3.00%) and 1, 3, 5-Araf (3.60%); it is inferred from the methylation results that the terminal arabinose (20.50%), galactose (9.80%), glucose (8.30%) and rhamnose (3.00%) constitute the side chain terminal of the polysaccharide, the main chain is composed of →4)-Galp-(1→ and →4)-Glcp-(1→ alternately; it is further inferred from the proportion and glycosidic bond connection mode that the main chain structure is →4)-Galp-(1→ and →4)-Glcp-(1→, with galactose as the main component and glucose as the auxiliary component, forming a linear skeleton connected by 1→4; the branching point of the polysaccharide is mainly →3, 6)-Galp-(1→, indicating that the galactose is simultaneously substituted at the 3 and 6 positions, and the secondary branching point is →3, 5)-Araf-(1→; the side chain terminal is dominated by Araf-(1→, indicating that arabinofuranose is the main side chain terminal, and there is a short chain or side chain in the form of →5)-Araf-(1→; the secondary branching is in the form of →3)-Galp-(1→ and →6)-Galp-(1→; the terminal residue accounts for 41.60%, indicating that it has a high degree of branching and belongs to a highly branched polysaccharide; at the same time, the main chain residue accounts for 20.60%, which forms a network structure in combination with the branching, and contains a high proportion of T-Araf and 1, 5-Araf, which is consistent with the characteristics of arabinogalactan.

[0145] Table 2 Analysis of methylation results of Kunlun baby's breath polysaccharide

[0146]

[0147] Test Example 5

[0148] Nuclear magnetic resonance determination of Kunlun baby's breath polysaccharide:

[0149] 30 mg of Kunlun polysaccharide was dissolved in deuterium oxide and freeze-dried, then dissolved in deuterium oxide and freeze-dried again, and the process was repeated three times to fully exchange the active hydrogen in the sample. The treated sample was dissolved in 0.5 mL of deuterium oxide and scanned on a nuclear magnetic resonance instrument for one-dimensional and two-dimensional spectra, and the results are shown in Figure 6 .

[0150] From Figure 6 , it can be seen that in the proton nuclear magnetic resonance spectrum, five characteristic resonance peaks are present in the region of 4.3 ppm-5.8 ppm, wherein the anomeric hydrogen signal follows the typical configuration distribution rule: the β-anomeric configuration (δ4.48 ppm, 4.66 ppm) is located in the interval of 4.3 ppm-5 ppm, and the α-anomeric configuration (δ5.12 ppm, 5.26 ppm, 5.80 ppm) is distributed in the high-field region of 5 ppm-5.8 ppm; as shown in Figure 6 , the simultaneous acquisition of 13 C spectrum, and Figure 6 , the HSQC two-dimensional correlation spectrum of D, further verifies the configuration characteristics of the anomeric carbon. The anomeric carbon signal presents five characteristic peaks in the range of 95 ppm-110 ppm, which are clearly related to the proton signal: δH / δC 4.48 / 101.19 (A), 4.66 / 103.59 (B), 5.12 / 99.47 (C), 5.26 / 109.70 (D), and 5.80 / 107.89 (E);

[0151] The signal assignment of each residue H and C is shown in Table 3. The δH 4.48 ppm, 4.66 ppm in residue A / B conforms to the anomeric hydrogen characteristics of β-pyranose, and the δC 101 ppm, 103 ppm is consistent with the anomeric carbon range of β-pyranose, the C4 shift of →4)-Glcp-(1→ is increased (~78.4 ppm), which is consistent with the glycosidic bond (2,3,6-Me3-Glcp) after methylation of polysaccharide, so the B residue is assigned as →4)-β-D-Glcp-(1→ 5] ; the C2 / C3 / C4 of Galp-(1→ terminal is not connected to other sugars, and the shift is low, so the residue A is assigned as β-D-Galp-(1→ 80]; δH > 5.0 ppm and δC > 99.0 ppm are in accordance with arabinofuranose; C5 linked to other sugars results in an up-shift of C5, residue C is assigned → 5)-a-L-Araf-(1→, a configuration anomeric hydrogen (5.26 ppm) and C1 (109.70 ppm) are in accordance with furanoid a-arabinose, residue D is assigned a-L-Araf-(1→. δH 5.80 ppm is the anomeric hydrogen characteristic peak of a-rhamnose (6-deoxy), slightly higher than typical pyranose due to deoxy effect and ring strain, chemical shift is higher than other pyranose, residue E is assigned a-D-Rhap-(1→. According to the methylation data, anomeric region signals and chemical shift information, the main chain of Kunlun baby's breath polysaccharide is composed of → 4)-β-D-Galp-(1→ and → 4)-β-D-Glcp-(1→ alternately (20.60%), embedded → 5)-a-L-Araf-(1→ flexible unit (11.50%); the branching point is located at → 3, 6)-β-D-Galp (9.60%), connecting two side chains - C3 position extending end a-L-Araf (20.50%), C6 position connecting β-D-Glcp / Galp end (8.30% / 9.80%); end a-D-Rhap (3.00%) modifies the side chain to form a highly branched heteropolysaccharide;

[0152] Therefore, the structure of Kunlun baby's breath polysaccharide is as follows:

[0153]

[0154] Table 3 Sugar residues in Kunlun baby's breath polysaccharide 1 H and 13 C signal assignment

[0155]

[0156] Test Example 6

[0157] To verify that Kunlun baby's breath polysaccharide enhances the hypoglycemic effect of metformin, the test arrangement is as shown in Table 4: Figure 7

[0158] Specifically, the test mice are divided into 4 groups, 10 mice in each group; streptozotocin-induced diabetic mice are divided into four groups: diabetic control group (marked as DC group), metformin group (marked as MET group), low-dose metformin and Kunlun baby's breath polysaccharide group (marked as LMET+AI group); the remaining 10 mice are allocated to the normal control group (NC);

[0159] The intragastric administration procedures of each group are as follows: the NC group and the DC group receive equal amounts of normal saline, the MET group receives 250 mg / kg of metformin, and the LMET+AI group receives 125 mg / kg of metformin and 200 mg / kg of Kunlun baby's breath polysaccharide. ​

[0160] Throughout the experiment, fasting blood glucose (FBG) levels were measured weekly and body weight was recorded. Bedding was changed daily to maintain a clean environment. After four weeks of treatment, all mice were fasted for 12 hours, weighed, and their feces were collected. Blood samples were collected by phoresis, placed in an ice bath for 1 hour, and then centrifuged at 4°C and 4000g for 10 minutes to collect serum. The serum was flash-frozen in liquid nitrogen and stored at -80°C. Some tissues and organs were fixed with 10% paraformaldehyde, and the remaining samples were stored at -80°C for further analysis.

[0161] Effects of LMET+AI on body weight and organs in diabetic mice:

[0162] Organ Index: After dissection, the pancreas, liver and kidneys are extracted, cleaned, dried and weighed. Organ Index = (organ weight) / (body weight);

[0163] Results: Streptozotocin-induced diabetic mice showed weight loss, swelling of organs such as the pancreas, increased organ indices, organ damage, and impaired organ function. Figure 8 As shown in Figures A and B, the NC group gradually increased its weight throughout the trial; conversely, after STZ induction, the model group experienced a significant decrease in weight; however, after intervention with MET or LMET+AI, the weight loss in these groups was significantly alleviated, with statistically significant differences compared to the DC group (p<0.05). Figure 8 As shown in C, D, and E, the pancreas, liver, and kidney indices in the NC group remained at the lowest levels (p<0.05); in contrast, the pancreas, liver, and kidney indices in the DC group increased due to STZ induction, leading to organ enlargement and elevated organ indices; however, after intervention with MET or LMET+AI, the organ indices tended to normalize; these results indicate that intervention with MET or LMET+AI can effectively reduce weight loss in diabetic mice and decrease elevated organ indices.

[0164] Effects of LMET+AI on glycemic index in diabetic mice:

[0165] Oral glucose tolerance test (OGTT): On day 26 of treatment, mice underwent an oral glucose tolerance test (OGTT) after fasting for 12 hours. Fasting blood glucose (FBG) levels were measured in each group using a Yuwell blood glucose meter (Danyang, China). Subsequently, each group was administered glucose solution by gavage at a dose of 2 grams per kilogram. Blood glucose levels were monitored at 30, 60, 90, and 120 minutes after administration.

[0166] Biochemical analysis: Glycated hemoglobin (HbA1c content) was determined using a kit from Beijing Bioscience and Technology Co., Ltd., serum insulin levels were determined using a kit from Nanjing Jiancheng Biological Engineering Institute, and liver and muscle glycogen content was evaluated using a glycogen detection kit from Nanjing Jiancheng Biological Engineering Institute.

[0167] Results: FBG reflects the blood glucose level of non-fasting mice and is an indicator of hyperglycemia. As shown in Table 4, the blood glucose level of the NC group was stable within the normal range (5.41 ± 0.31 mmol / L to 6.33 ± 0.85 mmol / L), confirming that there was no diabetes; in contrast, the FBG levels of the other groups were elevated, indicating that these mice were successfully induced to develop diabetes; after four weeks of intervention, the FBG levels of the MET and LMET+AI groups were significantly lower than those of the DC group (P < 0.05), indicating that these interventions had therapeutic effects in alleviating hyperglycemia.

[0168] OGTT is an indicator of the body's ability to handle glucose, and a hallmark of diabetes is that when the pancreas is impaired, glucose intolerance leads to elevated blood glucose. As shown in Table 5, the test mice underwent OGTT, and the results showed that the NC group exhibited normal glucose tolerance, while the DC group had significantly elevated blood glucose levels within 120 minutes of testing (p < 0.05), reflecting severe impairment of the pancreas; after treatment with MET or LMET+AI, the glucose tolerance of diabetic mice was improved; notably, LMET+AI was more effective than MET alone in improving glucose tolerance, indicating that Kunlun polysaccharides can enhance the therapeutic effect of MET.

[0169] Clinically, glycated hemoglobin (HbA1c) is commonly used as an indicator for monitoring diabetes, as shown in Table 6. Figure 8 As shown in Table 6, there were significant differences in HbA1c levels between groups (p < 0.05), with the lowest HbA1c level in the NC group, indicating stable blood glucose, and a significant increase in HbA1c in the DC group (p < 0.05); after treatment with MET and LMET+AI, the HbA1c levels of diabetic mice decreased, with the LMET+AI group showing greater improvement than the MET group; as shown in Table 6, there were also significant differences in insulin levels between groups (P < 0.05), with the lowest insulin level in the DC group, and both MET and LMET+AI treatment increased the insulin levels of diabetic mice, with LMET+AI being more effective than MET. Figure 8

[0170] Figure 9 ​Tables C and D show the results of glycogen content in mice. Glycogen synthesis involves converting glucose molecules into glycogen, which helps regulate blood sugar and provide energy during fasting or strenuous exercise. The NC group had higher levels of liver and muscle glycogen, indicating normal organ function. STZ-induced damage reduced glucose utilization, increased blood sugar, and decreased glycogen content. The LMET+AI group had higher muscle glycogen levels than the MET group (P < 0.05), indicating that Kunlun chrysanthemum polysaccharide can enhance the hypoglycemic effect of metformin. Kunlun chrysanthemum polysaccharide can serve as an adjuvant to improve the efficacy of metformin and reduce its dosage.

[0171] Table 4. Effects of Kunlun Chrysanthemum polysaccharide on FBG in diabetic mice

[0172]

[0173] Table 5. Effects of Kunlun Chrysanthemum polysaccharide on OGTT in diabetic mice

[0174]

[0175] Effects of LMET+AI on pancreatic tissue in diabetic mice:

[0176] H&E staining and immunofluorescence of pancreas: Pancreatic tissue was removed and immersed in fixative, processed into paraffin blocks and sectioned. Then, the paraffin on the sections was removed, stained, dehydrated and mounted. Microscopic examination and image capture were then performed. For frozen sections of pancreatic tissue, the tissue was first fixed and dehydrated (rinsed overnight in PBS and sucrose solution), then embedded in OCT medium and stored at -80°C. Sections were cut using a cryostat and stored at -80°C in preparation for immunofluorescence analysis.

[0177] Results: H&E staining was used to evaluate pancreatic histopathology, such as... Figure 10 As shown in Figure A, the DC group exhibited significant pancreatic islet atrophy, irregular shape, and reduced number of islet cells; the pancreatic tissue atrophy in the ME group and LMET+AI group of diabetic mice was significantly improved, indicating a significant repair effect on the pancreas; compared with the DC group, the pancreatic tissue of other groups showed a more regular and clear morphology, indicating that Kunlun tamarind polysaccharide can enhance the ability of MET to improve pancreatic damage in diabetic mice.

[0178] Cleavage Caspase-3 is the activated form of this cysteine ​​protease and a key mediator of apoptosis. Anti-apoptotic proteins can inhibit this process, such as... Figure 10As shown in FIG. 8B, immunofluorescence analysis showed that in STZ-induced mouse islets, cleaved Caspase-3 was widely co-localized with insulin, and the intervention of MET reduced this phenomenon, and the LMET+AI group was further improved, the co-staining of Caspase-3 and insulin was reduced, indicating that compared with MET alone, Kunlun polysaccharide can enhance the ability of MET to promote insulin secretion and reduce the expression of apoptosis-related proteins, thereby better protecting the pancreatic tissue;

[0179] NLRP-3 inflammasome is related to chronic inflammation associated with diabetes, especially in islet cells, such as Figure 10 As shown in FIG. 8C, compared with other groups, the DC group showed stronger NLRP-3 expression, but weaker insulin localization, and in the LMET+AI group, Kunlun polysaccharide assisted MET treatment reduced the inflammatory response of diabetic mice, while increasing insulin secretion; in the LMET+AI group and the MET group, the co-staining of NLRP-3 and insulin was reduced, but the improvement of pancreatic inflammation in the LMET+AI group was more obvious; these results show that Kunlun polysaccharide can enhance the effect of small-dose MET by inhibiting the cleavage of Caspase-3 and the inflammatory factor NLRP-3, thereby alleviating apoptosis and inflammation in diabetic mice.

[0180] Test Example 7

[0181] Further verify the hypoglycemic and hypolipidemic effect of Kunlun polysaccharide:

[0182] Construction of C. elegans high glucose and high fat model:

[0183] (1) Nematode genotype and culture conditions: the following nematode genotypes were selected: N2 Bristol (wild type), ldrls1 [dhs-3p::dhs-3::GFP] (Caenorhabditis Genetics Center); C. elegans was synchronized by using standard nematode growth medium (NGM) agar plates and fed with Escherichia coli OP50;

[0184] All tests were carried out at 20°C (unless otherwise specified);

[0185] (2) Establishment of high glucose and high fat model: C. elegans was exposed to 2% glucose NGM; NGM plates were used as controls (Control), 2% glucose NGM plates with different doses of AI-2 (0.1, 1 and 10 mg / mL), and 2% glucose NGM plates (Model); synchronized L1 C. elegans were grown under different dietary conditions at 20°C for 48 h until reaching the L4 stage;

[0186] Glycogen and oil red O staining analysis: C. elegans cultured under high sugar and high fat model, each group of nematodes were washed thoroughly with M9 buffer; using iodine staining method for glycogen staining to evaluate the effect of AI-2 (i.e. Kunlun baby's breath polysaccharide) on reducing blood sugar, L4 stage nematodes were observed by iodine vapor staining, and the effect of reducing blood sugar was observed; oil red O staining was used to evaluate the lipid-lowering effect of AI-2, and L4 stage nematodes were stained with oil red O, and the lipid accumulation was observed.

[0187] Results: Glycogen staining and oil red O staining can show the glucose content and fat content in vivo, and the expression intensity of the staining signal is analyzed to further evaluate the effect of AI-2 on reducing blood sugar and lowering lipid; as shown in Figure 11 (A), the glycogen staining results show that the glycogen in the Model group of nematodes is severely accumulated, indicating that the glucose content in vivo is high, indicating that the glucose content in the Model group is high; after AI-2 intervention, the glycogen content in the nematodes is reduced, which is consistent with the detection of glucose content. Due to the decrease of glucose content in vivo, the synthesis of glycogen is reduced. The signal intensity of glycogen staining is analyzed by ImageJ, and the glycogen signal of the Control group and the 10.0 mg / mL AI-2 group is the lowest, and the low, medium and high concentrations of AI-2 all show the effect of reducing blood sugar in nematodes. Figure 11 (B) can be seen that there is a significant difference in glycogen signal between groups (p<0.05), among which the glycogen signal of the Control group and the 10.0 mg / mL AI-2 group is the lowest, and the low, medium and high concentrations of AI-2 all show the effect of reducing blood sugar in nematodes. Figure 12 (A), the oil red staining of the nematodes in the Model group shows that it corresponds to the previously measured triglyceride, indicating that a large amount of fat is accumulated in vivo, and the nematodes remain in a high-fat state, and after AI-2 intervention, each concentration of AI-2 has the effect of reducing fat accumulation in nematodes; from Figure 12 (B) can be seen that the signal after oil red O staining is highly expressed in Model, and after AI-2 intervention, the trend is changed, and the oil red O signal expression in the high-fat model nematodes is reduced. From the above results, it can be seen that AI-2 can play a role in reducing blood sugar and lowering lipid.

[0188] Finally, it should be pointed out that: the above examples are only used to illustrate the technical solutions of the present application, but not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A Kunlun baby's chrysanthemum polysaccharide, characterized in that, mainly composed of arabinose, galactose, galacturonic acid, rhamnose and glucose; the main chain of the Kunlun baby chrysanthemum polysaccharide is composed of →4)-β-D-Galp-(1→ and →4)-β-D-Glcp-(1→ alternately; the side chain of the Kunlun baby chrysanthemum polysaccharide is composed of →5)-α-L-Araf-(1→, →3,6)-β-D-Galp, α-L-Araf, β-D-Glcp / Galp and α-D-Rhap; the molar percentage of the arabinose, galactose, galacturonic acid, rhamnose and glucose is 26.52:37.96:26.43:5.38:3.71; the structural formula of the repeating unit in the Kunlun baby chrysanthemum polysaccharide is as follows: α-L-Araf-(1→3) ↑ main chain: →4)-β-D-Galp-(1→4)-β-D-Glcp-(1→ | [→6)-β-D-Glcp-(1→] | α-L-Araf-(1→5)-α-L-Araf-(1→ | α-D-Rhap-(1→; the uronic acid content of the Kunlun baby chrysanthemum polysaccharide is 26.48±0.11%; the Kunlun baby chrysanthemum polysaccharide is an acidic polysaccharide with a molecular weight of 3192 Da; the preparation method of the Kunlun baby chrysanthemum polysaccharide comprises the following steps: (a) hydrolyzing the Kunlun baby chrysanthemum under acidic conditions, then filtering to obtain a filtrate; (b) concentrating the filtrate obtained in step (a), then performing alcohol precipitation, filtering to obtain a precipitate; (c) mixing the precipitate obtained in step (b) with a lye, then filtering, and adjusting the pH of the filtrate to neutral to obtain an extract; (d) performing ultrafiltration on the extract obtained in step (c) to obtain a filtrate and dry the filtrate to obtain a crude extract; (e) performing ultrafiltration on the crude extract obtained in step (d) to obtain a retentate and dry the retentate to obtain a Kunlun baby chrysanthemum crude polysaccharide; (f) treating the obtained Kunlun baby chrysanthemum crude polysaccharide by anion exchange chromatography and dialysis purification to obtain the Kunlun baby chrysanthemum polysaccharide; the method for hydrolyzing comprises the following steps: soaking the Kunlun baby chrysanthemum with acid, then hydrolyzing under the condition of cellulase synergistic magnetic field and electric field coupling to obtain a hydrolysis liquid; the stirring condition of the hydrolysis is stirring at 85 ℃ for 2h; the cellulase is cellulase with an enzyme activity of 800 U / g after addition, and the pH value is 5.3; the magnetic induction intensity of the magnetic field is 0.3T-0.5T; the alternating voltage frequency used by the electric field is 50Hz.

2. The Kunlun Scabious polysaccharide of claim 1, characterized in that, the filtering mode comprises centrifugation.

3. The Kunlun Scabious polysaccharide of claim 1, characterized in that, in step (c), the stirring condition of mixing the precipitate with the lye is stirring at 40℃-60℃ for 2h-3h.

4. The Kunlun Scabious polysaccharide of claim 1, characterized in that, in step (d), the molecular cut-off amount of the ultrafiltration is 10000 Da; in step (e), the molecular cut-off amount of the ultrafiltration is 5000 Da; in step (e), the molecular weight of the Kunlun baby chrysanthemum crude polysaccharide is 5000 Da-10000 Da.

5. The Kunlun Scabious polysaccharide of claim 1, characterized in that, The eluent used in the anion exchange chromatography comprises 0.01 mol / L NaOH solution and 3 times volume of 0 mol / L, 0.2 mol / L, 0.4 mol / L, 0.6 mol / L, 0.8 mol / L and 1.0 mol / L NaCl solution.

6. Use of the Kunlun baby's breath polysaccharide as claimed in any one of claims 1-5 in the preparation of a medicine for reducing blood sugar and / or reducing blood lipid.

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