CN114145457A

CN114145457A - Hypoglycemic composition

Info

Publication number: CN114145457A
Application number: CN202111504887.4A
Authority: CN
Inventors: 崔东影; 解庆刚; 刘莹; 石红丽; 蒋士龙; 冷友斌; 梁爱梅; 陈博
Original assignee: Heilongjiang Feihe Dairy Co Ltd
Current assignee: Heilongjiang Feihe Dairy Co Ltd
Priority date: 2021-12-10
Filing date: 2021-12-10
Publication date: 2022-03-08
Anticipated expiration: 2041-12-10
Also published as: CA3183772A1; CN114145457B; WO2023104181A1

Abstract

The invention relates to a functional nutritional composition, in particular to a blood sugar reducing composition. The composition comprises: i) one or more phytosterols; ii) one or more beta-glucans derived therefrom, wherein the composition has a content ratio of phytosterols to beta-glucans of 45:1 or less.

Description

Hypoglycemic composition

Technical Field

The invention belongs to the field of foods, relates to a functional nutritional composition, and more particularly relates to a functional health-care or nutritional composition with a blood sugar reducing effect.

Background

With the improvement of living standard and the change of life rhythm, the incidence of hypertension, hyperglycemia and hyperlipidemia is higher and higher, and the hypertension, hyperglycemia and hyperlipidemia are referred to. Belongs to cardiovascular and cerebrovascular diseases, and is mainly a 'rich disease' derived from modern civilization, which can exist alone or be related to each other. For example: diabetes patients are easy to suffer from hypertension or hyperglycemia which is a main factor for the formation and development of arteriosclerosis, and the elasticity of blood vessels of arteriosclerosis patients is poor, so that the increase of blood pressure is aggravated. Therefore, any of the three diseases is easy to develop and form 'three-high disease' in the later period, and the incidence rate is the highest especially among the old and the people with the hypofunction.

The number of people with diabetes is increasing every year from the global perspective and there is no obvious effective treatment. The incidence of diabetes also increases with age, with middle aged and elderly diabetics accounting for 98% of the total population of diabetics. Moreover, diabetes causes various complications, and the diabetes and the complications thereof can cause physical and mental injuries to patients, even threaten the life and health of the patients, and simultaneously cause dual losses of manpower and material resources to families of the patients. Therefore, preventing the occurrence of diabetes and alleviating the harm of diabetes are the problems which need to be solved urgently in our society nowadays. Nowadays, the society mostly adopts ways of diet control, oral administration of hypoglycemic drugs, insulin injection and the like to control blood sugar, which not only limits the diversity of food nutrient components ingested by patients, but also brings various pains and inconvenience to the patients. Therefore, the development of a healthy hypoglycemic composition without side effect is very important.

It is known that β -glucan is composed of glucose units, widely found in microorganisms, plants and animals, and is a polysaccharide unit of glucose. Structurally, most of the materials are combined by beta-1, 3 bonds, are polymers and have certain biological activity. In the middle of the 20 th century, foreign scholars found that a substance for enhancing immune function existed on the cell wall of yeast, and over 20 years later, the active substance was determined to be beta-1, 3-glucan. A plurality of experimental researches in recent years show that the beta-glucan has obvious effects on reducing blood sugar, reducing fat and improving immunity, and has great benefits on various physiological functions of a human body.

Yeast glucan, one of the sources of β -glucan, is derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae), has various physiological activities due to its specific structure, and has been classified as GRAS (generally recognized as safe) by FDA in the united states, and is widely used in various foods as a food raw material. And in most countries it is allowed to be used in all types of food. The yeast glucan as dietary fiber can be decomposed by intestinal bacteria, promotes gastrointestinal motility, stimulates secretion of digestive juice, and has the function of assisting digestion; the peristalsis of the intestinal tract can be increased, and the defecation is promoted, so that the constipation is effectively prevented; can increase the absorption of calcium, magnesium and zinc in human intestinal tract. The unique beta- (1 → 3) skeleton structure of the yeast glucan enables the yeast glucan to have stronger immunoregulation activity and can obviously improve the human immunity.

The phytosterol is recommended to be eaten by people with blood sugar abnormality for helping to reduce blood sugar and prevent cardiovascular diseases in multiple countries such as China, Europe, America and the like and medical association guidelines and consensus such as ESC/EAS and the like, and the recommended dose is 2-3 g/day.

Although some results show that β -glucan and phytosterols appear to have hypoglycemic efficacy, the biological demonstration of both is not sufficient (oat β -glucan intake between 3 and 8 weeks is reported to be probably beneficial for glycemic control in type 2 diabetic patients, but limited to small sample numbers, with no adequate conclusions). At present, the mixture of various substances with health care efficacy is most commonly used. For example:

citation 1 discloses a health food composition consisting of grains rich in plant polyphenols and sterols, traditional Chinese medicines and yeast. The invention exerts the synergistic hypoglycemic effect through the comprehensive action of various bioactive substance groups, and the synergistic effect of multi-level, multi-target and multi-angle such as polyphenol, sterol, dietary fiber, protein, polypeptide, oligopeptide, beta-glucan, flavone, D-chiral inositol, ecdysone and the like is realized, thereby showing excellent effect in the hypoglycemic experiment and clinical experiment of mice.

Citation 2 discloses a functional food for lowering blood sugar. The food comprises resistant starch, tea polyphenols and dietary fiber. The functional food has remarkable effect in inhibiting rapid increase of blood sugar, and has good daily health promotion effect for patients with diabetes and complications.

Citation 3 discloses a special clinical nutritional formula for gestational diabetes mellitus, which comprises protein, fat, carbohydrate, vitamins, medicinal and edible components, natural plant compounds and new resource food. Wherein the new resource food is selected from at least one of colostrum basic protein, gamma-aminobutyric acid, chitosan oligosaccharide, eucommia male flower, bamboo leaf flavone, casein phosphopeptide, oat beta-glucan, yeast beta-glucan, moringa leaves and xylo-oligosaccharide.

Citation 4 discloses a hypoglycemic pharmaceutical composition and a health functional food. The food is prepared from phytosterol ester, red yeast rice, tomato red and chromium-rich yeast, and tests prove that the food has a remarkable blood sugar reducing effect.

Although the above-mentioned prior art has already made some progress in reducing blood glucose by adding various functional ingredients, there is still room for further investigation for providing a functional composition for reducing blood glucose with a more definite mechanism and simpler ingredients.

Cited documents:

cited document 1: CN113575928A

Cited document 2: CN109602013A

Cited document 3: CN106666728A

Cited document 4: CN107095974A

Disclosure of Invention

Problems to be solved by the invention

As mentioned previously, various functional compositions have been proposed in the art for use in conjunction with glycemic control. However, in specific research practice, the inventors have also found the following problems:

the cited document 1 uses various plant materials directly, and does not examine the actual action and amount of the effective substances. As an optional component in cited document 2, it is proposed that one of various vitamins, sterols, amino acids, and β -glucan can be used, but there is no mention as to whether these components alone have a hypoglycemic effect. The cited reference 3 is complicated in composition, and although β -glucan is mentioned, it is not necessary as an essential component. Although the use of phytosterol is mentioned in the cited document 4, the whole body needs to have the hypoglycemic effect under the coordination of other various substances, and the relationship between specific synergistic effect components is not obvious.

These prior art references, however, do not mention the synergistic hypoglycemic effect of β -glucan and phytosterols and are generally used in higher amounts when phytosterols are used.

Therefore, in view of the above-mentioned disadvantages of the prior art, the present invention is directed to provide a composition having a hypoglycemic function with a simple composition, in which a synergistic effect can be obtained by combining beta-glucan and phytosterol in a limited amount, and experimental results show that the composition can obtain a significant hypoglycemic effect even when the amount of phytosterol or beta-glucan used is low.

In addition, the composition provided by the invention also has the effect of reducing blood fat.

Means for solving the problems

Through long-term research by the inventor of the present invention, the following technical solutions can be found to solve the above technical problems:

[1] the present invention provides, in a first aspect, a functional composition, wherein the composition comprises:

i) one or more phytosterols;

ii) one or more sources of beta-glucan,

wherein, in the composition, the dosage ratio of the phytosterol to the beta-glucan is below 45: 1.

[2] The composition according to [1], wherein the ratio of the phytosterol to the beta-glucan is 0.01:1 or more.

[3] The composition according to [1] or [2], wherein the phytosterol is selected from one or more of 4-desmethyl sterol, 4-methyl sterol and 4, 4' -dimethyl sterol.

[4] The composition according to any one of [1] to [3], wherein the beta-glucan is one or more selected from beta-glucans derived from plants or fungi.

[5] The composition according to any one of [1] to [4], wherein the content of the phytosterol is 0.1-10%; the content of the beta-glucan is 0.01-0.1%.

[6] The composition according to any one of [1] to [5], wherein the beta-glucan comprises yeast beta-glucan.

[7] The composition according to any one of [1] to [6], wherein the ratio of the phytosterol to the beta-glucan is 0.1-30: 1.

[8] The composition according to any one of [1] to [7], wherein the composition is a liquid or a semisolid.

[9] Further, the present invention provides a hypoglycemic composition, wherein the composition comprises the composition according to any one of [1] to [8].

[10] The present invention also relates to a food having both hypoglycemic and hypolipidemic effects, wherein the food is obtained from the composition according to any one of the above [1] to [9].

[11] The food according to [10], wherein the food comprises a pasta, a beverage, a reconstitutable food, a baked confectionery, a sauce or a functional nutritional supplement.

[12] The food product according to [11], wherein the beverage comprises a fruit or vegetable beverage, a tea beverage, a dairy beverage, a vitamin-based functional beverage.

ADVANTAGEOUS EFFECTS OF INVENTION

Through the implementation of the technical scheme, the invention has the following advantages and can obtain the following technical effects:

1) the functional composition for reducing blood sugar provided by the invention has simple and clear composition, is more beneficial to convenient and accurate compounding, and is also more beneficial to conveniently adding or preparing various foods, especially foods such as beverages;

2) the composition of the invention can obtain synergistic effect under the condition of the combination of the beta-glucan and the phytosterol with limited dosage, and the combination can also obtain obvious blood sugar reducing effect under the condition of lower dosage of the phytosterol;

3) the dosage of the beta-glucan and the phytosterol of the composition is within a safe dosage range, so that the composition can be used for a long time and is suitable for various unspecified people;

4) the composition disclosed by the invention has obvious blood sugar regulation and control effects, and also has the effects of assisting in reducing blood fat, resisting fatigue, improving immunity and the like.

Detailed Description

The present invention will be described in detail below. The technical features described below are explained based on typical embodiments and specific examples of the present invention, but the present invention is not limited to these embodiments and specific examples. It should be noted that:

in the present specification, the numerical range represented by "numerical value a to numerical value B" means a range including the end point numerical value A, B.

In the present specification, the numerical ranges indicated by "above" or "below" mean the numerical ranges including the numbers.

In the present specification, the meaning of "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

As used herein, the term "optional" or "optional" is used to indicate that certain substances, components, performance steps, application conditions, and the like are used or not used.

As used herein, "room temperature" means an indoor ambient temperature of "20 ℃.

In the present specification, the unit names used are all international standard unit names, and the "%" used means weight or mass% content, if not specifically stated.

In the present specification, the term "substantially" is used to indicate that the standard deviation from the theoretical model or theoretical data is within 5%, preferably 3%, and more preferably 1%.

In the present specification, reference to "some particular/preferred embodiments," "other particular/preferred embodiments," "embodiments," and the like, means that a particular element (e.g., feature, structure, property, and/or characteristic) described in connection with the embodiment is included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it is to be understood that the described elements may be combined in any suitable manner in the various embodiments.

< first aspect >

In a first aspect of the present invention, there is provided a functional composition comprising i) one or more phytosterols and ii) one or more beta-glucans derived from plant material, wherein the composition exhibits significant synergistic effects to significantly control and reduce blood glucose levels in humans, and also has immunity enhancing effects such as assisting in blood lipid reduction, fatigue resistance, and the like.

The technical scheme of the invention is mainly obtained based on the following insights:

the invention firstly verifies the safe dosage range of the phytosterol and the beta-glucan from the (cell) biology perspective, and respectively researches the respective biological performances of the phytosterol and the beta-glucan in the range and under the condition of different dosage combinations, so that the remarkable synergistic effect of the phytosterol and the beta-glucan on the aspect of reducing the blood sugar of a human body is unexpectedly found out under the specific content of the phytosterol and the beta-glucan.

(phytosterol)

Phytosterols (also known as phytosterols) are sterols present in higher plants. In clinical trials, phytosterols have been shown to block cholesterol absorption sites in the human gut and thus help reduce cholesterol levels in humans.

The relative density of phytosterol is slightly larger than that of water, the phytosterol is insoluble in water, acid and alkali, and the phytosterol can be dissolved in various organic solvents, such as vegetable fat, ether, benzene, chloroform, ethyl acetate, carbon disulfide and petroleum ether. Phytosterols are hydrophobic in nature, but they are hydrophilic in nature due to their hydroxyl groups, and therefore, phytosterols are generally emulsified to some extent.

The kind of phytosterol in the present invention is not particularly limited, and may be one or more of 4-desmethyl sterol, 4-methyl sterol, 4' -dimethyl sterol, and the like. In some specific embodiments, the phytosterols useful in the present invention may be selected from at least one of sitosterol, campesterol, stigmasterol, brassicasterol, and the corresponding alkanols, or mixtures thereof.

In addition, plant foods with high phytosterol content include vegetable oils, nuts and seeds, legumes, and the like. The phytosterol content in the vegetable oil is highest based on maize germ oil, and then the phytosterol content is sesame oil; the nut seeds have the highest content of pistachio nuts and are black sesame seeds; the soybeans have the highest content, and the soybeans are green beans; the phytosterol content in vegetables, fruits and potatoes is low.

In some specific embodiments of the present invention, plant materials containing phytosterols may be used for the phytosterols to impart a certain content of phytosterols in the final composition, and it is also preferable that the phytosterol active ingredient extracted from the plant with a higher purity may also be directly used.

Further, the mode of use of the extracted phytosterol is not particularly limited, and such a mixture of sterols may be used as it is, or the phytosterol may be used as it is in the form of a solution or emulsion.

(beta-glucan)

Beta-glucans (Beta-glucans for short), which belong to the class of fibrous polysaccharides (complex carbohydrates), are structurally largely a polymer bound by Beta-1, 3 bonds and can be of various microbial, animal or plant origin.

Specific sources of the β -glucan in the present invention are not particularly limited, and the sources of the β -glucan that can be suitably used in the present invention include one or a mixture of yeast, oat, barley, fungi (mushrooms), algae, and the like. More specifically, there may be mentioned, for example, oat β -glucan, kelp β -glucan, thermogelling β -glucan, golden alga kelp β -glucan, shiitake mushroom β -glucan, lichen β -glucan, oyster mushroom β -glucan, zymosan, black fungus β -glucan, and barley polysaccharide.

In some preferred embodiments of the invention, for the beta-glucan, one or a mixture of beta-glucans from yeast, oat, barley sources may be used, more preferably selected from yeast beta-glucan and/or oat beta-glucan.

In some specific embodiments of the present invention, plant, fungus and other raw materials containing the components can be used for the beta-glucan to impart a certain content of the beta-glucan in the final composition, and further preferably, the beta-glucan effective component extracted from these raw materials and having a higher purity can be directly used.

Further, the mode of using the extracted β -glucan is not particularly limited, and the β -glucan may be used as it is to form a mixture, or may be used as a solution or emulsion prepared from the β -glucan.

(synergistic)

Although the research on phytosterol and beta-glucan in the prior art has preliminary conclusions that both phytosterol and beta-glucan may have certain blood sugar regulation effect, on one hand, no research and report on whether both are present or may form synergistic blood sugar regulation capability exists, and on the other hand, most of the existing schemes use higher dosage to obtain the required effect when phytosterol is used, but no research and report on whether phytosterol can be matched with other components at lower content to play synergistic effect are available.

In the present invention, it is considered that the use of a combination of phytosterol and β -glucan at a specific content unexpectedly results in a synergistic effect on blood glucose control. Furthermore, it was also found that this synergistic effect also confers a blood glucose regulating effect which can be obtained also when using relatively low doses of phytosterol/β -glucan.

Specifically, in some particular embodiments of the invention, the ratio of phytosterols to beta-glucan may be 45:1 or less, preferably 30:1, more preferably 25:1 or less, most preferably 20:1 or less, and may be exemplified by (17, 18 or 19): 1 or less, the ratio of phytosterol is too high, and the synergistic effect of the two components may be poor. The lower limit of the ratio may be 0.01:1 or more, preferably 0.1:1 or more, more preferably 0.5:1 or more, and (0.3, 0.8, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15): 1 or more. If the ratio of the phytosterol is too low, there is a concern that the synergistic effect of the two components is not good in the case of a low absolute amount of the phytosterol.

Further, the amounts (especially, the maximum amounts) of phytosterol and β -glucan in the functional composition provided by the present invention are not limited in principle as long as the above-mentioned use ratio is satisfied. However, in consideration of food safety laws and regulations regarding different food types and the addition amounts of functional substances for different food types, in some specific embodiments, the content of the phytosterol may be 0.1% by mass or more, preferably 0.15 to 10% by mass, more preferably 0.2 to 6% by mass, still more preferably 1 to 5% by mass, and more preferably 2 to 3% by mass, based on the total mass of the composition, from the viewpoint of ensuring the synergistic effect and the safety of the use amount; the content of the β -glucan may be 0.01% by mass or more, preferably 0.01 to 0.2% by mass, more preferably 0.01 to 0.15%, even more preferably 0.01 to 0.1%, even more preferably 0.02 to 0.08%, and even more preferably 0.02 to 0.06% by mass. If the content is too low, the synergistic effect may be poor in some cases, and if the content is too high, the safety of the human body may be poor.

The invention further discovers that the effect of phytosterol or beta-glucan alone on regulating blood sugar is not obvious and sometimes even opposite to the effect of phytosterol or beta-glucan alone on ensuring cell activity. By matching the above contents, it is possible to exhibit a synergistic effect of controlling blood glucose even with a lower content of phytosterol or a lower content of β -glucan.

(composition)

The functional composition of the present invention comprises at least the phytosterol and beta-glucan as described above. The form of the composition is not particularly limited, and may be a mixture of the above-mentioned plants and fungi containing these components, or a mixture of two kinds of high-purity extracts.

The other components that can be used in the composition of the present invention are not particularly limited, and other edible ingredients, food additives, solvent components, and the like that are generally used in the art may be used without impairing the effects of the present invention.

The functional composition of the present invention may be present in a liquid form, or may be present in a semi-solid or solid form.

Furthermore, experiments prove that the composition has the function of improving the glucose intake reduction caused by PA, and the composition can reduce the insulin resistance induced by PA, so that the composition has a relatively obvious blood sugar regulation effect, and also has the effects of improving blood fat and improving human immunity, so that the composition can be used as a functional health product or a functional health additive.

< second aspect >

In a second aspect of the present invention, a functional health food, in particular a blood sugar controlling health food suitable for diabetic patients, is provided. Which is processed through a functional composition as described in accordance with the first aspect above.

The food of the present invention is not particularly limited, and may be a pasta, a beverage, a reconstitutable food, a baked confectionery, a sauce, or a functional nutritional supplement.

The pasta includes a staple food, a minor cereal staple food, and the like processed or produced from a flour-based raw material as a staple food, and specifically includes steamed bread, cakes, noodles, and stuffing-based staple food.

For the baked confectionery, there may be baked cake or biscuit or the like based mainly on cream, egg and fermented powder.

The beverage is not particularly limited, and may be a fruit beverage, a vegetable beverage, a milk tea beverage, a tea beverage, milk, yogurt, a vitamin beverage, or the like. In addition to the two functional components of the present invention, the fruit or vegetable beverage may further include juice of fruit or vegetable, or solid components thereof, and the like. The vitamin beverage may mainly contain various functional vitamins and other functional components in addition to the two functional components of the present invention, and specifically may include, for example, white granulated sugar, sodium cyclamate, acesulfame, taurine, potassium sorbate, lysine, inositol, vitamin pp, vitamin B6, vitamin B12, citric acid, and the like.

For reconstitutable foods, typically reconstitutable milk powder products may be used, such as adult milk powder, middle aged milk powder, and the like.

In addition, the functional nutritional supplement food is not particularly limited, and may be used as a nutritional supplement or as a meal replacement, and such food may include one or more of the following components in addition to the two functional components of the present invention: protein, fat, essential carbohydrates, dietary fiber, supplemental elements, vitamins, plant or dietary components, and the like.

Wherein the protein is at least one selected from whey protein powder, soy protein isolate, whole milk powder, whole egg powder, lactoferrin, bovine colostrum, amino acid and protein peptide; the amino acid is at least one selected from L-lysine-L-glutamic acid, L-arginine, L-tryptophan, L-glutamine, taurine, L-valine, L-isoleucine and L-leucine; the protein peptide is selected from one or more of soybean oligopeptide, wheat protein peptide, silkworm pupa protein peptide, marine fish oligopeptide powder, cola peptide, amino peptide and egg albumin peptide.

The fat may include at least one of saturated fatty acid, polyunsaturated fatty acid, monounsaturated fatty acid, OPO structural fat, DHA, EPA, ARA, and phospholipid, and more specifically the fat includes safflower oil, walnut oil, peanut oil, soybean oil, argan, olive oil, tea oil, mayberry oil, olive oil, coconut oil, perilla oil, deep sea fish oil, cocoa butter, palm oil, beef tallow, butter, lard, medium chain triglycerides, and lecithin.

The carbohydrate includes starch or modified starch. The dietary fiber comprises one or more of inulin, konjac flour, galacto-oligosaccharide, fructo-oligosaccharide, isomalto-oligosaccharide, soybean polysaccharide, cyclodextrin, resistant dextrin and soybean fiber.

The supplementary elements are selected from metal ion salts of organic acids such as calcium citrate, calcium L-lactate, calcium hydrogen phosphate, potassium gluconate, sodium citrate, ferrous gluconate, potassium iodide, zinc gluconate, sodium selenite, copper gluconate, chromium sulfate, manganese gluconate, and magnesium gluconate.

The vitamins are selected from one or more of vitamin A, beta-carotene, vitamin D3, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, pantothenic acid, folic acid, nicotinic acid, choline, inositol and biotin.

The plant or dietary component comprises one or more of cereals, beans, potatoes, vegetables, bacteria and algae, nuts and seeds, fish and shrimp, livestock meat, and fruits.

Examples

Hereinafter, the present invention will be further specifically described by way of specific examples. The reagents, methods and apparatus employed in the present invention are those conventional in the art, unless otherwise specified.

< raw materials >

In the examples section that follows, the starting materials used are as follows, unless otherwise indicated.

Phytosterol: shanghai-derived leaf Biotechnology Ltd, S26734

Yeast beta glucan: shanghai Tongyuan food technology Co., Ltd, HH1006

In the following sections, unless otherwise indicated, when referring to parts by mass and ratios of ingredients in a nutritional composition, the parts by mass refer to parts by mass of phytosterol, yeast beta glucan as active ingredients, and the ratios refer to mass ratios of phytosterol, yeast beta glucan as active ingredients.

Each experimental example was prepared by mixing the phytosterol, yeast beta glucan starting material according to the composition shown in table 1 below.

Table 1:

< cell experiment example >

1. Materials and methods

1.1. Experimental reagent and consumable

See the following table for experimental main reagents and sources:

table 2-1:

1.2. experimental equipment

The main experimental instruments and equipment of the invention are shown in the following table:

tables 2 to 2:

numbering	Name of instrument	Company(s)
			1	Carbon dioxide incubator	Thermo corporation of America
2	Biological safety cabinet	Thermo corporation of America
			3	Enzyme-linked immunosorbent assay (ELISA) instrument	Dynatech Inc. of USA
4	Palm type centrifuge	Haimaicheilin Beel instruments Ltd
			5	Desk type high-speed refrigerated centrifuge	Beckman, Germany
6	Inverted microscope	Japan OLMPUS Corp
			7	Magnetic stirrer	Hangzhou instruments ltd
8	Vortex mixer	Haimaicheilin Beel instruments Ltd
			9	Constant temperature water bath	Shanghai Sensin laboratory instruments Ltd
10	Real-time fluorescent quantitative PCR	Bio-Rad Inc. of USA

1.3. Experimental methods

1.3.1 cell culture

HepG2 cells were placed in DMEM complete medium containing 10% FBS, 37 ℃, 5% CO₂Culturing in a saturated humidity cell culture box, and changing the culture solution every other day. And (3) passage can be carried out when the confluency is more than 80% in the logarithmic growth phase, the cells can be washed for 2 times by PBS, the cells are digested by 0.25% trypsin for 10-15 min until the cells are separated from the bottle wall, the cells are transferred to a centrifuge tube, a sterile needle is blown to single cells, and the cells are subjected to passage culture according to the proportion of 1: 3. Cells in the logarithmic growth phase were taken for the experiments.

1.3.2 cell viability assay

Preparing single cell suspension of HepG2 liver cells, mixing about 1X 10⁴The individual cells were seeded in 96-well plates and cultured until the cells accounted for approximately 80% of the wells for drug intervention. After 24h of drug intervention, 10 μ L of CCK8 solution was added to each well of the plate using a pipette. After incubating in a 37 ℃ incubator for 4 hours, absorbance at 450nm was measured using a microplate reader.

1.3.3 preparation of Long-chain fatty acid esters

Dissolving Palmitic Acid (PA) in 20mL of absolute ethyl alcohol, adding 1.6mol/L of sodium hydroxide after fatty acid is completely dissolved in the absolute ethyl alcohol, saponifying the mixture, drying the saponified semi-solidified solution in a fume hood for 48 hours to obtain a precipitate, placing the precipitate in a 80 ℃ water bath kettle after the precipitate is completely dried, heating the solution to be completely dissolved until the temperature of the solution is reduced to about 50 ℃, adding 20% (w/v) BSA (bovine serum albumin) with the same volume as the solution for conjugate dissolution, placing the solution in a 50 ℃ magnetic stirrer for magnetic stirring for 4 hours to completely combine the fatty acid with the BSA to form fatty acid-BSA (the preparation concentration is 3mmol/L), and filtering and sterilizing the fatty acid ester by using a 0.22 mu m filter membrane for later use.

1.3.4 cell glucose consumption assay

The medicine is intervened in the cells for 2 hours in advance, then 0.2mM Palmitate (Palmitate, PA) is added for treatment for 24 hours, culture solution supernatant is sucked, glucose consumption conditions are reflected by measuring glucose content (Beijing prilley glucose consumption measuring kit) of the culture solution supernatant, measurement is carried out according to kit instructions, the insulin resistance degree of the cells is fed back through glucose consumption, the lower the glucose consumption is, the higher the insulin resistance degree is, the higher the glucose consumption is, the insulin resistance degree is reduced.

1.4. Cell assay grouping and dosing

The specific experimental doses are shown in tables 2-3 below.

Tables 2 to 3:

1.5 statistical analysis

All experiments were repeated three times independently, statistical analysis was performed on the experimental data using SPSS 22.0 statistical software, and the experimental results were examined for homogeneity of variance, and statistical differences between groups were analyzed using one-way variance, expressed as mean ± standard deviation (x ± SD). P >0.05 means no statistical significance, and P <0.05 means difference is statistically significant.

2. Results of the experiment

2.1. Screening of each sample for a safe dose of HepG2 cells

Tables 2-4 show the effect of yeast beta glucan on HepG2 cell viability; tables 2-5 show the effect of phytosterols on the viability of HepG2 cells.

Tables 2 to 4:

note:^**represents the ratio of P < 0.01 to the control group;^***represents the ratio of P < 0.001 to the control group;

tables 2 to 5:

note: because the water solubility of the phytosterol is poor, the phytosterol is prepared from ethanol.

The CCK-8 cell proliferation detection kit is a rapid sensitivity kit which is based on WST-8 (chemical name: 2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2, 4-disulfophenyl) -2H-tetrazole monosodium salt) and is widely applied to cell proliferation and cytotoxicity detection. In the presence of an electron-coupling agent, WST-8 can be reduced by dehydrogenase in the mitochondria to produce a highly water-soluble orange-yellow formazan product (formazan), the shade of which is proportional to the proliferation of viable cell numbers and inversely proportional to cytotoxicity. The number of the living cells can be indirectly reflected by measuring the OD value at the wavelength of 450nm by using a microplate reader, and the higher the detected OD value is, the more the number of the living cells is, and the lower the OD value is, the less the number of the living cells is.

2.2 Effect of Each sample on insulin resistance of HepG2 cells

To measure the effect of mixed nutrition or nutrient-alone treatment on PA-induced insulin resistance of HepG2 cells, we evaluated the proportion of cellular glucose consumption to assess the effect of improving blood glucose. Tables 2-6 show the effect on insulin resistance of different nutritional composition samples comprising yeast beta glucan, phytosterols

Tables 2 to 6:

note:^*p is less than 0.05 compared with the blank control group,^**p is less than 0.01 compared with the blank control group;^∧compared with the positive control group, p is less than 0.05,^∧∧compared with the positive control group, p is less than 0.01,^∧∧∧p is less than 0.001 compared with positive control group

The cell insulin resistance is reduced due to insulin resistance, and the cell insulin resistance degree is reflected by detecting the cell glucose consumption, so that the cell insulin resistance degree is high when the cell glucose consumption is low, and the cell insulin resistance degree is low when the cell glucose consumption is high.

The sample treatment results showed that phytosterols and yeast beta glucan when used alone, exhibited no or limited effect in reducing PA-induced insulin resistance while ensuring cellular activity, as compared to the model control group. However, when the two are used in combination, especially the phytosterol: when the metering ratio of the yeast beta glucan is controlled to be below 45:1, the effect of obviously reducing PA-induced insulin resistance can be obtained.

Specifically, as shown by comparing experimental example 1, experimental example 4 and experimental example 7, cellular experimental examples 1 and 4 respectively increase the proportion of intracellular glucose consumption induced by PA by 0.000 and-0.005, and cellular experimental example 7 increases the proportion of intracellular glucose consumption by 0.016, which is greater than the sum of the two (0.000+ -0.005), compared with the cellular experimental control group, which indicates that a synergistic effect exists between the two components, and the synergistic effect can synergistically increase the proportion of glucose consumption and assist in reducing blood glucose. Meanwhile, the data also show that the low-dose phytosterol and the low-dose yeast beta glucan can also have good effects of improving insulin resistance and assisting in reducing blood sugar.

The invention has the beneficial effects that: the invention provides the influence of the composition of the phytosterol and the yeast beta glucan on reducing blood sugar, particularly on improving PA-induced insulin resistance, and provides a new idea for the development of future functional foods. The phytosterol and yeast beta glucan have wide prospects in improving blood sugar. Our studies show that in an in vitro model, HepG2 cells treated with a combination of yeast beta glucan and phytosterol showed a significant improvement in insulin resistance with a good synergistic effect between the components.

< application example >

In the following application examples, the "parts" are parts by weight, and the% contents of the components are% by weight.

In addition, in each application example below, the following raw material sources were used as follows unless otherwise specified.

Raw milk: heilongjiang Feihe Dairy Co Ltd

Whole milk powder: heilongjiang Feihe Dairy Co Ltd

Phytosterol ester: qinhuang island Jinhai Special edible oil industry Co Ltd

Yeast beta glucan: shanghai Tongyuan food technology Co., Ltd, HH1006

Compounding vitamins: dismaman vitamin (Shanghai) Co Ltd

Compounding minerals: dismaman vitamin (Shanghai) Co Ltd

Solid corn syrup bowling-Bao biological shares Ltd

Lactose: hilmar USA

Skim milk powder: ireland Kerry

Isomaltooligosaccharide: bowling Bao biological shares Ltd

Mono-diglycerol fatty acid ester: danisco (China) Ltd

Application example 1

The modified milk powder containing the phytosterol and the yeast beta glucan is suitable for middle-aged and elderly people, and each 1000 parts of the modified milk powder is prepared from the following components in parts by weight:

the milk powder of the invention uses the following raw materials: 436 parts of full cream milk powder, 8.08 parts of phytosterol ester (99% of phytosterol in each part), 0.51 part of yeast beta glucan (98.0% of yeast beta glucan in each part), 250 parts of skim milk powder, 83.41 parts of lactose, 179 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 2 parts of compound vitamin and 1 part of compound mineral; mixing the above materials, pasteurizing, homogenizing, evaporating for concentration, spray drying to obtain semi-finished powder, and packaging the mixed milk powder with nitrogen to obtain the final product. The product contains 0.8% of phytosterol and 0.05% of yeast beta glucan in a ratio of 16: 1.

Application example 2

the milk powder of the invention uses the following raw materials: 436 parts of full cream milk powder, 12.12 parts of phytosterol ester (99% of phytosterol in each part), 0.26 part of yeast beta glucan (98.0% of yeast beta glucan in each part), 250 parts of skim milk powder, 79.62 parts of lactose, 179 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 2 parts of compound vitamin and 1 part of compound mineral; mixing the above materials, pasteurizing, homogenizing, evaporating for concentration, spray drying to obtain semi-finished powder, and packaging the mixed milk powder with nitrogen to obtain the final product. The product contains 1.2% of phytosterol and 0.025% of yeast beta glucan in a ratio of 48: 1.

Application example 3

the milk powder of the invention uses the following raw materials: 436 parts of full cream milk powder, 2.42 parts of phytosterol powder (99% of phytosterol in each part), 0.61 part of yeast beta glucan (98.0% of yeast beta glucan in each part), 250 parts of skim milk powder, 88.97 parts of lactose, 179 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 2 parts of compound vitamin and 1 part of compound mineral; mixing the above materials, pasteurizing, homogenizing, evaporating for concentration, spray drying to obtain semi-finished powder, and packaging the mixed milk powder with nitrogen to obtain the final product. The product contains 0.24% of phytosterol and 0.06% of yeast beta glucan in a ratio of 4: 1.

Application example 4

The modified milk containing the phytosterol and the yeast beta glucan is prepared from the following components in parts by weight per 1000 parts of the modified milk:

the modified milk comprises 990.29 parts of raw milk, 0.51 part of yeast beta glucan (98.0 percent of yeast beta glucan in each part), 0.40 part of phytosterol powder (99 percent of phytosterol in each part), 8.3 parts of fructo-oligosaccharide and 0.5 part of monoglyceride and diglycerol fatty acid ester; mixing the above materials, homogenizing, UHT sterilizing, homogenizing, and aseptic packaging to obtain the final product. The product contains 0.04% of phytosterol and 0.05% of yeast beta glucan in a ratio of 0.8: 1.

Application example 5

the modified milk comprises 983.04 parts of raw milk, 0.61 part of yeast beta glucan (98.0 percent of yeast beta glucan in each part), 0.29 part of phytosterol powder (99 percent of phytosterol in each part), 15.56 parts of fructo-oligosaccharide and 0.5 part of monoglyceride and diglycerol fatty acid ester; mixing the above materials, homogenizing, UHT sterilizing, homogenizing, and aseptic packaging to obtain the final product. The product contains phytosterol 0.029%, yeast beta glucan 0.06%, and the ratio is 0.48: 1.

Application example 6

The modified milk containing the phytosterol and the yeast beta glucan is suitable for middle-aged and elderly people, and each 1000 parts of the modified milk is prepared from the following components in parts by weight:

the milk powder of the invention uses the following raw materials: 436 parts of full cream milk powder, 12.12 parts of phytosterol powder (99% of phytosterol in each part), 0.51 part of yeast beta glucan (98.0% of yeast beta glucan in each part), 250 parts of skim milk powder, 79.37 parts of lactose, 179 parts of solid corn syrup, 40 parts of isomaltooligosaccharide, 2 parts of compound vitamin and 1 part of compound mineral; mixing the above materials, pasteurizing, homogenizing, evaporating for concentration, spray drying to obtain semi-finished powder, and packaging the mixed milk powder with nitrogen to obtain the final product. The product contains 1.2% of phytosterol and 0.05% of yeast beta glucan in a ratio of 24: 1.

Industrial applicability

The functional composition provided by the invention can be industrially prepared and is suitable for regulating and controlling blood sugar.

Claims

1. A functional composition, wherein the composition comprises:

i) one or more phytosterols;

ii) one or more sources of beta-glucan,

2. The composition of claim 1, wherein the ratio of phytosterols to β -glucan is 0.01:1 or greater.

3. A composition according to claim 1 or 2, wherein the phytosterols are selected from one or more of 4-desmethyl sterols, 4-methyl sterols and 4, 4' -dimethyl sterols; the beta-glucan is selected from one or more of beta-glucan derived from plants or fungi.

4. The composition according to any one of claims 1 to 3, wherein the content of the phytosterol is 0.1% by mass or more; the content of the beta-glucan is 0.01 mass% or more.

5. A composition according to any one of claims 1 to 4, wherein the β -glucan comprises yeast β -glucan.

6. The composition according to any one of claims 1 to 5, wherein the ratio of the phytosterol to the beta-glucan is 0.1-30: 1.

7. A composition according to any one of claims 1 to 6, wherein the composition is a liquid or semi-solid.

8. A hypoglycemic composition comprising a composition according to any one of claims 1 to 7.

9. A food product obtainable from a composition according to any one of claims 1 to 8.

10. The food product of claim 9, wherein the food product comprises a pasta, beverage, reconstitutable food, baked confectionery, sauce, or functional nutritional supplement, and the beverage comprises a fruit or vegetable beverage, a tea beverage, a dairy beverage, or a vitamin functional beverage.