CN116115731A - Composition and preparation for reducing blood sugar and application of composition and preparation - Google Patents

Abstract

The invention provides a hypoglycemic composition, a preparation and application thereof. The composition comprises spirulina peptide, broccoli polysaccharide and at least two selected from beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide and tannic acid. The composition has good effect of reducing blood sugar and treating hyperglycemia, and can delay the development of hyperglycemia complications.

Description

Composition and preparation for reducing blood sugar and application of composition and preparation

Technical Field

The invention relates to the technical field of biological medicines, in particular to a composition and a preparation for reducing blood sugar and application thereof.

Background

Diabetes is a global public health problem today. Among the numerous pathogenic mechanisms, glycosylated end products (AGEs) are being studied intensively as an important member of them. The advanced glycosylation end product refers to a kind of chemical substances which are generated by the reaction of reducing sugar with substances such as protein and the like under the condition of non-enzymatic reaction, have stable and irreversible properties and special absorption spectrum and fluorescence characteristics and have damage. The pathological mechanism of the AGEs is probably that the AGEs directly cross-link with intracellular and extracellular proteins to influence the structure and the function of the proteins, so that the organs and tissues are damaged. And damage does not stop after AGEs are formed, and various AGEs such as hemoglobin-AGEs, lens proteins-AGEs, collagen-AGEs and the like can also continue to move around, continue to damage other tissue cells, and also serve as a continuous activation of the immune system. These AGEs are very difficult to metabolize by the human body, and stay in the body for a very long time, which also aggravates the damage of AGEs to the human body, and these AGEs can continue to accelerate the occurrence and development of various diseases. Anti-glycosylation, anti-oxidation is therefore particularly important for the prevention of complications.

In addition, the pathogenic process of AGEs contains a large number of oxidation-reduction reactions, and oxygen free radicals are needed to participate. After AGEs bind to their Receptors (RAGE), oxidative stress is induced to generate a large number of oxygen radicals, thereby activating the transcription factor Nuclear Factor (NF) -KB, which is sensitive to oxygen radicals. NF-KB is a pleiotropic transcriptional regulator of a variety of "damage response genes", whose activation can cause expression of such genes in target cells, inducing the production of a variety of damage factors, producing pathogenic effects.

Beta-alanine is a potent AGEs inhibitor; beta-alanine not only inhibits the production of AGEs, but also binds to and inactivates already formed AGEs.

The paeonol can not only remove free radicals in the initiation stage of the chain reaction in the oxidation resistance reaction, but also directly capture the free radicals in the chain of the free radical reaction, and the blocking of the chain reaction of the free radicals plays a double role in preventing and breaking the chain. Not only can inhibit the generation of AGEs, but also can combine with the formed AGEs to inactivate the AGEs.

After the dendrobium chrysotoxum root crude polysaccharide is orally taken, the dendrobium chrysotoxum root crude polysaccharide is absorbed by the gastrointestinal tract, has strong oxidation resistance and anti-glycation activity, and blocks non-enzymatic glycosylation and protein crosslinking toxic reaction induced by reactive aldehyde.

The green persimmon tannic acid extract has the most powerful function of resisting saccharification, and the anti-saccharification meaning that sugar molecules are not combined with proteins in a human body, so that the effect is mainly to keep blood sugar in a known stable state and avoid diabetes.

The broccoli refined polysaccharide has strong DPPH oxygen free radical scavenging ability and can block lipid peroxide generation. In addition, the broccoli refined polysaccharide has the function of regulating blood lipid, can reduce the content of triglyceride and Low Density Lipoprotein (LDL), and further reduces the formation of AGEs.

Spirulina (spirorina) is a blue algae with the highest protein content in various foods at present, and is widely applied to the fields of health-care foods, medicines and the like. In recent years, spirulina peptides have been found to have various effects as important active substances of spirulina. Can inhibit angiotensin converting enzyme I, and has blood pressure lowering effect. In addition, it has inhibiting effect on pancreatic lipase and 3T3-L1 preadipocyte, and can reduce triglyceride accumulation. Although biological activities of spirulina peptides such as antioxidation, antibiosis, lipid lowering and the like are widely studied at home and abroad, blood sugar lowering activity research of spirulina peptides in a type 2 diabetes model has not been reported yet.

Disclosure of Invention

In order to solve the technical problems in the prior art, the invention provides a composition which has the efficacy of reducing the glycosylation end product, can be used for treating hyperglycemia and can delay the development process of hyperglycemia complications.

In a first aspect, the present invention provides a hypoglycemic composition comprising a spirulina peptide, a broccoli polysaccharide and at least two selected from the group consisting of beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid.

The invention discovers that the combination of the spirulina peptide, the broccoli polysaccharide and at least two components selected from beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide and tannic acid has better blood sugar reducing effect, and the synergistic blood sugar reducing effect among a plurality of components has obvious effect especially in reducing AGEs of glycosylation end products.

In some embodiments, the composition comprises a spirulina peptide, a broccoli polysaccharide, and any two selected from the group consisting of β -alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid.

In some preferred embodiments, the composition comprises dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide.

In some preferred embodiments, the composition comprises paeonol, tannic acid, spirulina peptides, and broccoli polysaccharides.

In some preferred embodiments, the composition comprises paeonol, dendrobium chrysotoxum polysaccharide, spirulina peptide and broccoli polysaccharide.

In some embodiments, the composition comprises a spirulina peptide, a broccoli polysaccharide, and any three selected from the group consisting of beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid.

In some preferred embodiments, the composition comprises paeonol, dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide.

In some more preferred embodiments, the composition comprises beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide.

In some embodiments, the mass percent of beta-alanine is 1-3%, e.g., 1%, 1.2%, 1.5%, 1.8%, 2%, 2.2%, 2.5%, 2.8%, 3%, or any value therebetween, based on the total mass of the composition.

In some embodiments, the mass percent of paeonol is 2-5%, e.g., 2%, 2.2%, 2.5%, 2.8%, 3%, 4%, 4.2%, 4.5%, 4.8%, 5%, or any value therebetween, based on the total mass of the composition.

In some embodiments, the dendrobium chrysotoxum polysaccharide is present in an amount of 0.1-0.3% by mass, e.g., 0.1%, 0.12%, 0.15%, 0.18%, 0.2%, 0.22%, 0.25%, 0.28%, 0.3% or any value therebetween, based on the total mass of the composition.

In some embodiments, the tannic acid is present in an amount of 0.1 to 0.2% by mass, e.g., 0.1%, 0.12%, 0.15%, 0.18%, 0.2% or any value therebetween, based on the total mass of the composition.

In some embodiments, the spirulina peptide is present in an amount of 0.1-0.5% by mass, e.g., 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5% or any value therebetween, based on the total mass of the composition.

In some embodiments, the percentage by mass of the broccoli polysaccharide is 3-6%, e.g., 3%, 3.2%, 3.5%, 3.8%, 4%, 4.2%, 4.5%, 4.8%, 5%, 5.2%, 5.5%, 5.8%, 6% of any value therebetween, based on the total mass of the composition.

In some embodiments, the composition comprises, based on 100% total weight of the composition, dendrobium chrysotoxum polysaccharide 0.1-0.3%, tannic acid 0.1-0.2%, spirulina peptide 0.1-0.5% and broccoli polysaccharide 3-6%.

In some embodiments, the composition comprises 2-5% paeonol, 0.1-0.2% tannic acid, 0.1-0.5% spirulina peptide and 3-6% broccoli polysaccharide, based on 100% total weight of the composition.

In some embodiments, the composition comprises 2-5% paeonol, 0.1-0.3% dendrobium chrysotoxum polysaccharide, 0.1-0.5% spirulina peptide and 3-6% broccoli polysaccharide, based on 100% total weight of the composition.

In some embodiments, the composition comprises 2-5% paeonol, 0.1-0.3% dendrobium chrysotoxum polysaccharide, 0.1-0.2% tannic acid, 0.1-0.5% spirulina peptide and 3-6% broccoli polysaccharide, based on 100% total weight of the composition.

In some embodiments, the composition comprises, based on 100% total weight of the composition, 1-3% beta-alanine, 2-5% paeonol, 0.1-0.3% dendrobium chrysotoxum polysaccharide, 0.1-0.2% tannic acid, 0.1-0.5% spirulina peptide, and 3-6% broccoli polysaccharide.

Preferably, water is also included in the composition.

In some embodiments, the composition consists of, based on 100% total weight of the composition: 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

In some embodiments, the composition consists of, based on 100% total weight of the composition: 2-5% of peony phenol, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

In some embodiments, the composition consists of, based on 100% total weight of the composition: 2-5% of peony phenol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

In some preferred embodiments, the composition consists of, based on 100% total weight of the composition: 2-5% of peony phenol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

In some more preferred embodiments, the composition consists of, based on 100% total weight of the composition: 1-3% of beta-alanine, 2-5% of peony phenol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

The present invention is not particularly limited to the respective raw materials, and can be prepared by using a conventional method in the art.

In some embodiments, the beta-alanine is obtained by a bio-enzymatic method. In some embodiments, the β -alanine of the present invention can be prepared by the following method:

adding acrylic acid into a reaction vessel, adjusting the pH to 8.0-8.5 by using ammonia water, adding beta-alanine catalytic enzyme, and stirring and reacting at 48-52 ℃ to obtain the beta-alanine.

In some embodiments, the paeonol is a polyphenol extracted from peony. In some embodiments, the paeonol may be prepared via an alcohol extraction process:

(1) Pulverizing flos moutan, and mixing with 1g of the following materials: (5-15) adding methanol as an extractant into the mixture, oscillating the mixture for 24-36 hours at room temperature, separating supernatant, and drying the supernatant to obtain a methanol extract, wherein the concentration of the methanol is preferably 70%;

(2) Dispersing the methanol extract in distilled water according to the feed liquid ratio of 1g (25-35) mL, then adding n-butane according to the volume ratio of 1 (3-8), extracting, and collecting the aqueous phase extract;

(3) Mixing the water phase extract collected in the step (2) with diethyl ether according to the volume ratio of 1 (3-5), extracting, and collecting the water phase extract;

(4) Mixing the water phase extract collected in the step (3) with ethyl acetate in a volume ratio of 1 (5-7), extracting, collecting the ethyl acetate extract, concentrating and drying to obtain the paeonol extract.

In some embodiments, the dendrobium chrysotoxum polysaccharide is a polysaccharide extracted from dendrobium chrysotoxum root, preferably, the dendrobium chrysotoxum polysaccharide is dendrobium chrysotoxum polysaccharide dry powder. In some embodiments, the dendrobium chrysotoxum polysaccharide of the present invention can be prepared by the following method:

(1) Crushing dendrobium chrysotoxum roots, and adding ethanol according to the volume ratio of 1 (4-6) to obtain a mixture;

(2) Filtering the mixture obtained in the step (1), preferably filtering with nylon net to obtain filtrate, adding 2-3 times of methanol into the filtrate, and heating and reflux extracting to obtain extractive solution;

(3) Standing the extractive solution, drying the precipitate, grinding, and sieving to obtain polysaccharide dry powder.

In some embodiments, the tannic acid is tannic acid extracted from green persimmon. For example, tannic acid of the present invention can be prepared by the following method:

(1) Crushing green persimmons, homogenizing, taking 10-20g of homogenized pulp, and adding hydrochloric acid methanol solution according to a feed liquid ratio of 1g (5-15) mL to obtain slurry; preferably, the volume ratio of the hydrochloric acid-methanol solution is (1:100) in terms of HCl;

(2) Performing ultrasonic extraction on the slurry, and performing suction filtration on the extract to obtain a filter cake;

(3) Eluting with absolute ethanol until the eluate and ferric trichloride-potassium ferricyanide do not react blue, concentrating the eluate, and freeze drying to obtain polysaccharide crude extract.

In some embodiments, the spirulina peptide is a polypeptide obtained from spirulina by enzymatic methods. In some embodiments, the spirulina peptides of the invention can be prepared by the following method:

centrifuging fresh spirulina after ultrasonic crushing, and freeze-drying to obtain spirulina peptide;

adding complex enzyme and alkaline protease into spirulina peptide, and reacting to obtain a solution containing spirulina peptide; preferably, the temperature of the reaction is 40-50 ℃ and the reaction time is 4-8h;

the solution containing the spirulina peptide passes through an ultrafiltration membrane and is freeze-dried to obtain a spirulina peptide finished product; the molecular weight cut-off of the ultrafiltration membrane is 1500-2500Mw.

In some embodiments, the broccoli polysaccharide is a polysaccharide extracted from broccoli by an alcohol extraction method, preferably the broccoli polysaccharide is a broccoli polysaccharide dry powder. In some embodiments, the broccoli polysaccharide of the present invention may be prepared by the following method:

(1) Drying broccoli, pulverizing, and sieving;

(2) Degreasing with petroleum ether (3-8) mL, adding ethanol, and removing monosaccharide, oligosaccharide and pigment; preferably, the concentration of the ethanol is 70-80%;

(3) Concentrating under reduced pressure, adding ethanol, dialyzing, vacuum filtering, and drying to obtain refined polysaccharide of herba Cephalanoploris; preferably, the concentration of ethanol is 90-99%.

In a second aspect, the present invention provides a hypoglycemic agent comprising a composition according to the first aspect and acceptable excipients.

Preferably, the auxiliary materials are selected from one or more of microcrystalline cellulose, starch, micro silica gel, hydroxypropyl methyl cellulose, xylitol, lactose, sucralose, edible essence and sucrose fatty acid ester.

The preparation of the invention can be solution, emulsion, suspension, tablet, powder, granule or capsule.

In a third aspect, the present invention provides the use of a composition according to the first aspect for reducing blood glucose.

Preferably, the composition is used for preparing a hypoglycemic drink, food, health care product or medicine.

The beneficial effects of the invention are as follows: the composition disclosed by the invention combines spirulina peptide, broccoli polysaccharide, beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid and the like, has a good blood sugar reducing effect, particularly has a remarkable effect in inhibiting the generation of AGEs (glycation end products), and can reduce the generation of hyperglycemia complications and delay the development process of the complications. The components of the invention also have synergistic hypoglycemic effect.

Drawings

FIG. 1 shows the results of detection of AGEs, FBG and HbAIc of the compositions of the control group and examples 1 to 7 and comparative examples 1 to 2.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, examples and drawings are provided below to aid in understanding the present invention. It is to be understood that these examples and drawings are for illustrative purposes only and are not to be construed as limiting the invention in any way. The actual scope of the invention is set forth in the following claims. It will be understood that any modifications and variations may be made without departing from the spirit of the invention.

Example 1

The composition with the hypoglycemic effect comprises the following components in percentage by mass: 0.2% of dendrobium chrysotoxum root crude polysaccharide, 0.1% of green persimmon tannic acid extract, 0.1% of spirulina peptide, 3% of broccoli refined polysaccharide and the balance of water.

Example 2

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: 2% of peony phenol, 0.1% of green persimmon tannic acid extract, 0.1% of spirulina peptide, 3% of broccoli refined polysaccharide and the balance of water.

Example 3

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: 2% of peony phenol, 0.2% of dendrobium chrysotoxum root crude polysaccharide, 0.1% of spirulina peptide, 3% of broccoli refined polysaccharide and the balance of water.

Example 4

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: 5% of peony phenol, 0.2% of dendrobium chrysotoxum root crude polysaccharide, 0.1% of green persimmon tannic acid extract, 0.3% of spirulina peptide, 6% of broccoli refined polysaccharide and the balance of water.

Example 5

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: beta-alanine 3%, peony phenol 3%, dendrobium chrysotoxum root crude polysaccharide 0.2%, green persimmon tannic acid extract 0.1%, spirulina peptide 0.3%, broccoli refined polysaccharide 5% and the balance of water.

Example 6

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: beta-alanine 1%, peony phenol 2%, dendrobium chrysotoxum root crude polysaccharide 0.2%, green persimmon tannic acid extract 0.2%, spirulina peptide 0.5%, broccoli refined polysaccharide 6% and the balance of water.

Example 7

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: beta-alanine 0.1%, peony phenol 0.2%, dendrobium chrysotoxum root crude polysaccharide 0.1%, green persimmon tannic acid extract 0.2%, spirulina peptide 3%, broccoli refined polysaccharide 0.1%, and the balance being water.

Comparative example 1

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: 5.6% of spirulina peptide, 6% of broccoli refined polysaccharide and the balance of water.

Comparative example 2

A composition with the effect of reducing glycosylation end products, which comprises the following components in percentage by mass: 5% of beta-alanine, 5.5% of peony phenol, 0.5% of dendrobium chrysotoxum root crude polysaccharide, 0.6% of green persimmon tannic acid extract and the balance of water.

The components and mass ratios of the compositions in the above examples and comparative examples are shown in Table 1.

TABLE 1

The components in the above examples were prepared as follows:

1. the beta-alanine is prepared by an enzyme catalysis method:

adding acrylic acid into a reaction vessel, adjusting the pH to 8.0-8.5 by using ammonia water, adding beta-alanine catalytic enzyme, and stirring and reacting at 48-52 ℃ to obtain the beta-alanine.

2. The peony phenol is prepared by the following steps:

(a) Selecting deep color peony, shearing and crushing, wherein the granularity is 300-500 microns, and the ratio of feed to liquid is 1g: adding 10ml of 70% methanol as an extracting agent, oscillating for 24-36h at room temperature, separating supernatant, and drying the supernatant at low temperature to obtain methanol extract with a yield of 5% -16%;

(b) Dispersing the methanol extract in distilled water with the feed-liquid ratio of 1g to 30ml, adding n-butane according to the volume ratio of 1:3-8, oscillating and extracting for 4-5 times, and collecting the aqueous phase extract;

(c) Adding diethyl ether into the aqueous phase extract with the volume ratio of 1:3-5, oscillating and extracting for 2-3 times, and collecting the aqueous phase extract;

(d) Adding ethyl acetate into the extract obtained in the step (c) in a volume ratio of 1:5-7, carrying out vibration extraction for 2-3 times, collecting the ethyl acetate extract, concentrating and drying at a low temperature to obtain the paeonol extract, wherein the yield is 5-10%.

3. The dendrobium chrysotoxum root crude polysaccharide is prepared by the following method:

(a) Crushing dendrobium chrysotoxum roots, and adding ethanol according to the volume ratio of 1:5;

(b) Filtering with nylon net to obtain filtrate, adding 2 times of methanol, heating and reflux extracting, repeating for 3 times, and mixing filtrates;

(c) Standing the filtrate for more than 6 hours, pouring out the supernatant, drying the precipitate, crushing and grinding, and sieving with a 100-mesh sieve to obtain polysaccharide dry powder.

4. The green persimmon tannic acid is extracted by the following method:

(a) Crushing green persimmons, homogenizing, taking 15g of homogenized pulp, filling into a beaker, and adding 2% hydrochloric acid methanol solution according to a feed-liquid ratio of 1:10;

(b) Performing ultrasonic extraction for 3 times, mixing the extracts, standing for more than 6 hours, and performing suction filtration to obtain a filter cake;

(c) Eluting with absolute ethanol until the eluate and ferric trichloride-potassium ferricyanide do not react blue, concentrating the eluate, and freeze drying to obtain crude extract.

5. The spirulina peptide is prepared by the following method:

centrifuging fresh spirulina after ultrasonic crushing, freeze-drying to obtain spirulina peptide, reacting spirulina peptide solution containing 3% complex enzyme (the volume ratio of protein mass to pure water is 1:30, pH 7.0) at 45 ℃ for 6h, inactivating at 95 ℃ for 5min, adding 6% alkaline protease, and reacting at 45 ℃ for 6h; and then the spirulina peptide product is obtained after the spirulina peptide product is freeze-dried through a 2000Mw ultrafiltration membrane.

The fresh spirulina used in the preparation is provided by Lijiang Cheng Baol biological development Co., ltd; the complex enzyme is prepared from cellulase (Ctec 2) and pectase (Pectinex Ultra SP-L) according to a weight ratio of 2:1, and is provided by Norwestin biotechnology Co., ltd; alkaline protease (S-ALPT) is supplied by Beijing Soy Biotechnology Co.

6. The used broccoli refined polysaccharide is prepared by the following method:

(a) Drying 500g of broccoli to constant weight, crushing and sieving with a 80-mesh sieve;

(b) Degreasing with petroleum ether 1:5, removing monosaccharide, oligosaccharide and pigment with 75% ethanol, and repeating for 3 times;

(c) Concentrating under reduced pressure with rotary evaporator, adding 95% ethanol as dialysate, dialyzing for 48 hr, vacuum filtering, and drying to obtain refined polysaccharide.

And (3) blood sugar reducing effect test:

1. establishment and grouping of diabetic rat models

260 clean grade healthy male SD rats (about 200g in weight, provided by the laboratory animal center of the university of Western An traffic medical college) were raised at room temperature of 20-24 ℃ and air relative humidity of 50% -70%, cycled for 12 hours at day and night, and adapted for one week.

After one week of adaptive feeding, SD rats were randomly divided into three groups; each group of 20.

Streptozotocin (Sigma) was dissolved in 0.1mmol/L Citrate Buffer (CB) at pH4.2 and freshly prepared as a 38mmol/L STZ solution immediately before use. Rats in the experimental group were fasted for 12 hours and given 50mg/kg streptozotocin for a single intraperitoneal injection; the normal control group was injected with the same dose of citrate buffer. Taking trace blood from the rat tail after 72h of intraperitoneal injection and one week to measure Fasting Blood Glucose (FBG), wherein the two Fasting Blood Glucose (FBG) is more than 16.7mmol/L, and urine glucose is determined as experimental animal, and diabetes model establishment is determined, and the patient with the blood glucose is removed with the blood glucose less than 16.7 mmol/L. The model animals were randomly subdivided into four groups of 20 animals each.

The administration method comprises the following steps: rats were monitored weekly for blood glucose and body weight after molding. Two weeks later, one, two, three, four, five, six, seven groups of the compositions of examples 1-7 were administered by drenching with a 1ml/100g weight; the compositions of comparative examples 1 to 2 were administered in equal amounts to each of the comparative experiment one and comparative experiment two groups, and the control group was administered with the same volume of physiological saline. For a total of 12 weeks. All rats were fed in separate cages, were fed ad libitum, and were illuminated with natural light around the clock.

2. Specimen collection

All rats were placed in metabolic cages over 12 weeks, fasted without water withdrawal, and 24h urine was left. All rats were anesthetized with 80mmol/L sodium pentobarbital and 6mL of heart blood was collected after centrifugation at 3500r/min for 15 min.

3. Serum FBG detection, hbAIc detection and AGEs measurement

The full-automatic biochemical analyzer is Beckmann Coulter AU5800 full-automatic biochemical analyzer; the HbAIc ELISA kit used was supplied by Beijing Aoweiya Biotechnology Co., ltd;

(1) Fasting glycation (FBG) was measured using a fully automatic biochemical analyzer to test the anti-glycation ability of the samples.

(2) Glycosylated hemoglobin (HbAIc) was tested using a HbAIc ELISA kit to test anti-glycation ability of a sample, which was performed strictly according to the ELISA kit instructions.

(3) Glycosylated end products (AGEs) were detected using a hitachi 650-60 fluorescence spectrophotometer to test the ability of the samples to reduce glycosylated end products.

(4) Fluorescence measurement of AGEs: the fluorescence values were determined on a Hitachi 650-60 fluorescence spectrophotometer (PBS zeroed, excitation wavelength 370nm, slit 5nm, emission wavelength 440 rim). Purified AGE-BSA was diluted with PBS to various concentrations (0, 1g/mL,5g/mL,10g/mL,50g/mL,100 g/mL) and used as a standard, a standard curve was drawn from its fluorescence values, an equation was fitted, and the AGEs value (F-AGEs) was calculated from the equation, 1 unit of F-AGEs corresponding to 1g/mLAGE-BSA.

4. And (3) data processing:

the results of serum FBG detection, hbAIc detection and AGEs measurement in step 3 are shown in fig. 1 and table 2.

Examples 1-7 changes in serum FBG and glycosylated hemoglobin in rats: compared with comparative examples 1-2 and control group, the serum AGEs data of rats of model examples 1, 2, 3, 4, 5, 6 and 7 were significantly reduced, as were the FBG and HbAIc values; example 5 has the most remarkable effect.

TABLE 2

Group of	Number of examples	AGEs(KU/L)	FBG(mmol/mL)	HbAIc(μg/mL)
					Control group	20	27.39±1.59	35.058±4.715	45.566±4.563
Example 1	20	19.98±1.02	31.359±3.416	38.235±3.569
					Example 2	20	18.25±1.32	30.732±2.578	37.366±1.952
Example 3	20	18.27±0.89	31.098±2.365	38.062±1.072
					Example 4	20	15.25±0.78	28.256±1.586	36.265±1.065
Example 5	20	9.34±0.85	25.364±1.655	33.955±1.389
					Example 6	20	11.25±0.86	26.36±1.678	34.965±1.523
Example 7	20	18.254±0.45	30.954±1.871	38.546±4.521
					Comparative example 1	20	22.515±0.25	32.155±1.555	41.932±4.541
Comparative example 2	20	21.154±0.58	32.944±2.245	42.552±3.647

The differences were statistically significant (P < 0.05), but the comparative differences between groups were not statistically significant (P > 0.05).

From the data in table 2 and fig. 1, it can be seen that the composition of the final products of glycosylation provided in examples 1-7 of the present invention can well inhibit the production of AGEs of the final products of glycosylation by combining β -alanine, paeonol, crude polysaccharide of dendrobium chrysotoxum root, green persimmon tannic acid extract, spirulina peptide, and refined polysaccharide of broccoli, thereby achieving the purpose of controlling blood sugar.

Example 4 compared with example 5, in the case that the total amount of the active components is unchanged, in example 5, beta-alanine is used together with paeonol, dendrobium chrysotoxum root crude polysaccharide, green persimmon tannic acid extract, spirulina peptide, broccoli refined polysaccharide and the like, so that the hypoglycemic effect of the composition can be obviously improved, and the beta-alanine has better synergistic effect with paeonol, dendrobium chrysotoxum root crude polysaccharide, green persimmon tannic acid extract, spirulina peptide and broccoli refined polysaccharide, wherein the beta-alanine has unexpected effect of reducing glycosylation end products.

The data of comparative example 1 and comparative example 2 show that only spirulina peptide and broccoli refined polysaccharide or only beta-alanine, paeonol, dendrobium chrysotoxum root crude polysaccharide and green persimmon tannic acid extract in the active ingredients of the composition can not obviously reduce glycosylation end products, and the aim of the test can not be achieved. The data of comparative example 1 and example 5 show that the use of beta-alanine, paeonol, crude dendrobium chrysotoxum root polysaccharide and green persimmon tannic acid extract together with spirulina peptide and broccoli refined polysaccharide in example 5 can greatly improve the blood sugar reducing effect under the condition that the total amount of active components is unchanged, and the synergistic blood sugar reducing effect is also shown between the beta-alanine, the paeonol, the crude dendrobium chrysotoxum root polysaccharide and green persimmon tannic acid extract and the spirulina peptide and the broccoli refined polysaccharide.

AGEs values for example 5 were significantly lower than for the control group; and the FBG value and HbAIc value are lower than those of the control group, which shows that the effect of the embodiment 5 is most remarkable, and the improvement of the capacity of reducing the glycosylation end product is realized from three aspects of AGEs, FBG and HbAIc.

The technical scheme of the invention is not limited to the specific embodiment, and all technical modifications made according to the technical scheme of the invention fall within the protection scope of the invention.

Claims (10)

1. A blood sugar reducing composition comprises spirulina peptide, broccoli polysaccharide and at least two selected from beta-alanine, paeonol, herba Dendrobii polysaccharide and tannic acid.

2. The composition of claim 1, wherein the composition comprises a spirulina peptide, a broccoli polysaccharide, and any two selected from the group consisting of β -alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid;

preferably, the composition comprises dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide; or alternatively

The composition comprises paeonol, tannic acid, spirulina peptide and broccoli polysaccharide; or alternatively

The composition comprises paeonol, dendrobium chrysotoxum polysaccharide, spirulina peptide and broccoli polysaccharide.

3. The composition according to claim 1 or 2, wherein the composition comprises spirulina peptide, broccoli polysaccharide and any three or four selected from the group consisting of β -alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid;

preferably, the composition comprises paeonol, dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide;

more preferably, the composition comprises beta-alanine, paeonol, dendrobium chrysotoxum polysaccharide, tannic acid, spirulina peptide and broccoli polysaccharide.

4. A composition according to any one of claims 1-3, characterized in that the mass percentage of β -alanine is 1-3% based on the total mass of the composition; and/or

Based on the total mass of the composition, the mass percentage of the paeonol is 2-5%; and/or

Based on the total mass of the composition, the mass percentage of the dendrobium chrysotoxum polysaccharide is 0.1-0.3%; and/or

The tannic acid is 0.1-0.2% by mass based on the total mass of the composition; and/or

The mass percentage content of the spirulina peptide is 0.1-0.5% based on the total mass of the composition; and/or

The content of the broccoli polysaccharide is 3-6% by mass based on the total mass of the composition.

5. The composition of any one of claims 1-4, wherein the composition comprises, based on 100% total weight of the composition, dendrobium chrysotoxum polysaccharide 0.1-0.3%, tannic acid 0.1-0.2%, spirulina peptide 0.1-0.5% and broccoli polysaccharide 3-6%; or alternatively

Based on 100 percent of the total weight of the composition, the composition comprises 2-5 percent of paeonol, 0.1-0.2 percent of tannic acid, 0.1-0.5 percent of spirulina peptide and 3-6 percent of broccoli polysaccharide; or alternatively

Based on 100 percent of the total weight of the composition, the composition comprises 2 to 5 percent of paeonol, 0.1 to 0.3 percent of dendrobium chrysotoxum polysaccharide, 0.1 to 0.5 percent of spirulina peptide and 3 to 6 percent of broccoli polysaccharide; or alternatively

Based on 100 percent of the total weight of the composition, the composition comprises 2 to 5 percent of paeonol, 0.1 to 0.3 percent of dendrobium chrysotoxum polysaccharide, 0.1 to 0.2 percent of tannic acid, 0.1 to 0.5 percent of spirulina peptide and 3 to 6 percent of broccoli polysaccharide; or alternatively

Based on the total weight of the composition as 100%, the composition comprises 1-3% of beta-alanine, 2-5% of paeonol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide and 3-6% of broccoli polysaccharide.

6. The composition of any one of claims 1-5, wherein the composition further comprises water,

preferably, the composition consists of the following components, based on 100% of the total weight of the composition:

2-5% of peony phenol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water; or alternatively

The composition consists of the following components by taking the total weight of the composition as 100 percent:

1-3% of beta-alanine, 2-5% of peony phenol, 0.1-0.3% of dendrobium chrysotoxum polysaccharide, 0.1-0.2% of tannic acid, 0.1-0.5% of spirulina peptide, 3-6% of broccoli polysaccharide and the balance of water.

7. The composition of any one of claims 1-6, wherein the β -alanine is obtained by a bio-enzymatic method; the peony phenol is a polyphenol substance extracted from peony by an alcohol extraction method; the dendrobium chrysotoxum polysaccharide is polysaccharide extracted from dendrobium chrysotoxum roots; the tannic acid is tannic acid extracted from green persimmon; the spirulina peptide is polypeptide obtained from spirulina by an enzymolysis method; the broccoli polysaccharide is a polysaccharide extracted from broccoli by an alcohol extraction method.

8. A hypoglycemic formulation comprising the composition of any one of claims 1-7 and an adjuvant;

preferably, the auxiliary materials are selected from one or more of microcrystalline cellulose, starch, micro silica gel, hydroxypropyl methyl cellulose, xylitol, lactose, sucralose, edible essence and sucrose fatty acid ester.

9. The formulation of claim 8, wherein the formulation is a solution, emulsion, suspension, tablet, powder, granule, or capsule.

10. Use of a composition according to any one of claims 1-7 for lowering blood sugar, preferably for the preparation of a beverage, food, health product or pharmaceutical product for lowering blood sugar.

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