JP7162542B2 - Food composition for promoting food factor sensing-related gene expression and food factor sensing-related gene expression promoter - Google Patents

Description

TECHNICAL FIELD The present invention relates to a food composition for promoting food factor sensing-related gene expression and a food factor sensing-related gene expression promoter .

In recent years, awareness of health has been increasing year by year, and research on health has been actively conducted in the food field as well. Food has a primary function of sustaining life by supplying nutrients, a secondary function of satisfying taste and taste, and a tertiary function of regulating physiological systems in the body to maintain and restore health. . In recent years, this tertiary function has attracted attention, and many foods for specified health uses, foods with nutrient function claims, and foods with function claims have been developed.

As foods for specified health use, many foods claiming effects such as suppressing increases in blood pressure and blood sugar levels and suppressing absorption of sugar and body fat are sold. However, many of the details of the mechanism of action of functional ingredients contained in food for specified health use have not been elucidated.

In recent studies on the mechanism of action of functional food ingredients, it has been reported that functional food ingredients bind to intracellular and extracellular receptors and exert their functions after binding (Non-Patent Document 1). This means that the functional ingredients of food bind to specific target molecules to transmit signals into cells and exert their effects. However, the results show that the effects are different.

On the other hand, Euglena (genus name: Euglena, Japanese name: Euglena) is attracting attention as a biological resource that is expected to be used as food, feed, fuel, and the like.
Euglena contains 59 types of nutrients, including vitamins, minerals, amino acids, and unsaturated fatty acids, which correspond to most of the nutrients necessary for human life. It has been proposed for its use and possible use as a food source in poor areas where necessary nutrients cannot be obtained.

Euglena is located in the primary producer of the food chain, and it is preyed on by predators, and culture conditions such as light, temperature conditions, and agitation speed are difficult compared to other microorganisms. However, in recent years, the intensive research of the present inventors has established a mass culture technique, opening the way for mass supply of Euglena and Euglena-derived substances such as paramylon extracted from Euglena.
Euglena is a unique organism that possesses chloroplasts as a plant and performs photosynthesis while having the animal-like nature of flagellum movement. Many functionalities of Euglena itself and substances derived from Euglena have been reported. (Non-Patent Document 2). Many aspects of the functionality and mechanism of action in the body of Euglena itself and Euglena-derived substances have not yet been elucidated in detail, and there is a demand for the creation of new functionality and the elucidation of the mechanism of action in the body.

H. Tachibana, et al., Nat. Struct. Mol. Biol., vol.11, pp.380-381 (2004). Ryo Arashida, "Characteristics of microalgae Euglena and their applications in food and environmental fields", Photosynthesis Research, vol.22, no.1, pp.33-38 (2012).

The present invention has been made in view of the above problems, and an object of the present invention is to enable daily and continuous ingestion and to effectively promote the expression of food factor sensing-related genes. It is an object of the present invention to provide a food composition for promoting food factor sensing-related gene expression and an agent for promoting food factor sensing-related gene expression .

As a result of intensive studies aimed at solving the above problems, the present inventors have found that Euglena-derived substances have the effect of promoting the expression of genes related to food factor sensing, and have completed the present invention.

Therefore, according to the present invention, the above object is to contain Euglena as an active ingredient, promote the expression of Ppara (SEQ ID NO: 17) , and enhance the lipid metabolism improving action, which is the food factor function of unsaturated fatty acids. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

In addition, according to the present invention, the above-mentioned object is achieved by containing Euglena as an active ingredient, which is used to promote the expression of Vdr (SEQ ID NO: 3 5 ), and enhances the bone protective effect, which is the food factor function of vitamin D. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

In addition, according to the present invention, the above-mentioned problem is solved by containing euglena as an active ingredient, using it to promote the expression of Ffar1 (SEQ ID NO: 9), and enhancing the antidiabetic action, which is the food factor function of long-chain fatty acids. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

Further, according to the present invention, paramylon is contained as an active ingredient, used to promote the expression of Sirt1 (SEQ ID NO: 37), and is used to enhance the anti-aging effect, which is the food factor function of flavonoids. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

Further, according to the present invention, paramylon is contained as an active ingredient, is used to promote the expression of Gabbr1 (SEQ ID NO: 10), and is used to enhance the blood pressure lowering action that is the food factor function of GABA. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

Further, according to the present invention, there is provided paramylon, which contains paramylon as an active ingredient, is used to promote the expression of Adra1a (SEQ ID NO: 3), and enhances the adrenergic action, which is the food factor function of syneferin. It is solved by a food composition for promoting food factor sensing-related gene expression, which is characterized by being used for

In addition, according to the present invention, the above-mentioned problem is achieved by containing Euglena as an active ingredient, promoting the expression of Ppara (SEQ ID NO: 17), and enhancing the lipid metabolism improving action, which is the food factor function of unsaturated fatty acids. It is solved by a food factor sensing-related gene expression promoter characterized by being used.

In addition, according to the present invention, the above-mentioned object is achieved by containing Euglena as an active ingredient, which is used to promote the expression of Vdr (SEQ ID NO: 3 5 ), and enhances the bone protective effect, which is the food factor function of vitamin D. It is solved by a food factor sensing-related gene expression promoter characterized by being used for
In addition, according to the present invention, the above-mentioned problem is solved by containing euglena as an active ingredient, using it to promote the expression of Ffar1 (SEQ ID NO: 9), and enhancing the antidiabetic action, which is the food factor function of long-chain fatty acids. It is solved by a food factor sensing-related gene expression promoter characterized by being used for
Further, according to the present invention, paramylon is contained as an active ingredient, used to promote the expression of Sirt1 (SEQ ID NO: 37), and is used to enhance the anti-aging effect, which is the food factor function of flavonoids. It is solved by a food factor sensing-related gene expression promoter characterized by being used for.
Further, according to the present invention, paramylon is contained as an active ingredient, is used to promote the expression of Gabbr1 (SEQ ID NO: 10), and is used to enhance the blood pressure lowering action that is the food factor function of GABA. It is solved by a food factor sensing-related gene expression promoter characterized by being used for.
Further, according to the present invention, there is provided paramylon, which contains paramylon as an active ingredient, is used to promote the expression of Adra1a (SEQ ID NO: 3), and enhances the adrenergic action, which is the food factor function of syneferin. It is solved by a food factor sensing-related gene expression promoter characterized by being used for.

According to the present invention, a novel food composition for promoting food factor sensing-related gene expression, a food factor sensing-related gene expression promoter, a food composition for enhancing food factor function, an agent for enhancing food factor function, and a food for enhancing food factor function Additives can be provided.
The food composition for promoting food factor sensing-related gene expression, the food factor sensing-related gene expression promoter, the food composition for enhancing food factor function, the food factor function enhancing agent, and the food additive for enhancing food factor function of the present invention are There have been no reports of side effects until now, and the active ingredient is a Euglena-derived substance with a level of safety that meets the Food Sanitation Law, so long-term continuous administration and continuous intake are possible.

1 is a graph showing changes in gene expression levels in the liver due to ingestion of Euglena-derived substances. 1 is a graph showing changes in gene expression levels in the liver due to ingestion of Euglena-derived substances. 1 is a graph showing changes in gene expression levels in the liver due to ingestion of Euglena-derived substances. 1 is a graph showing changes in gene expression levels in the liver due to ingestion of Euglena-derived substances. FIG. 2 shows food factor sensing-related genes whose expression is elevated in the liver. 1 is a graph showing changes in gene expression levels in quadriceps femoris muscles due to ingestion of Euglena-derived substances. 1 is a graph showing changes in gene expression level in the quadriceps femoris muscle due to ingestion of a Euglena-derived substance. 1 is a graph showing changes in gene expression level in the quadriceps femoris muscle due to ingestion of a Euglena-derived substance. FIG. 3 shows food factor sensing-related genes whose expression is elevated in the quadriceps femoris muscle. FIG. 2 is a conceptual diagram for explaining an example of Euglena's effect of enhancing food factor functions. FIG. 2 is a conceptual diagram for explaining an example of the food factor function-enhancing effect of paramylon. Fig. 3 is a graph showing the measurement results of Esr1 and Esrrb gene expression levels in the quadriceps femoris muscle (Data are means ± SE, n = 5, Student's t-test, *p < 0.05 vs. control). Fig. 10 is a graph showing the measurement results of the Ctsd gene expression level in the quadriceps femoris muscle (Data are means ± SE, n = 5, Student's t-test, *p < 0.05 vs. control).

Embodiments of the present invention will be described below with reference to FIGS.
This embodiment includes a food composition for promoting food factor sensing-related gene expression containing an Euglena-derived substance as an active ingredient, a food factor sensing-related gene expression promoter, a food composition for enhancing food factor function, a food factor function enhancer, and The present invention relates to food additives for enhancing food factor functions.

<Food factor>
A food factor is a group of substances that play an important role in the tertiary function of food, namely, "physiological regulation function", and is synonymous with "functional food factor".
Food factors (functional food factors) are physiologically active substances that regulate physiological systems in the body and have effects on maintenance and recovery of health.

Although the classification of functional food factors is not uniform, there are classifications by food components, classifications by target diseases, classifications based on the place of action (biological system), classifications based on the mechanism of action, etc. (Science Editing Committee of Food Functionality, "Science of Food Functionality", April 2008).

(Classification by food ingredients)
Examples of classification by food component are shown below for each food component group, but functional food factors are not limited to those shown below.
・Carbohydrates: indigestible polysaccharides, oligosaccharides, etc. ・Lipids: polyunsaturated fatty acids, plant sterols, glycerophospholipids, sphingolipids, etc. ・Proteins: functional proteins such as milk proteins and soybean proteins, oligopeptides, amino acids, etc.・Vitamins: vitamin E (tocopherol, tocotrienol), vitamin K (menaquinone), folic acid, etc. ・Minerals: calcium, magnesium, iron ・Nucleotides: AMP (adenylic acid), CMP (cytidylic acid), GMP (guanylic acid), IMP ( inosinic acid)
・Carotenoids: carotene, lycopene ・Isoprenoids: terpenes ・Polyphenols: flavonoids, isoflavones, anthocyanidins ・Sulfur-containing compounds: isothiocyanate ・Microorganisms: bifidobacteria, lactic acid bacteria

(Classification by target disease)
As examples of classification by target disease, examples of functional food factors for each disease are shown below, but functional food factors are not limited to those shown below.
・High blood pressure: peptides, geniposidic acid, acetic acid, n-3 fatty acids ・Hypercholesterolemia: soybean protein ・peptides, chitosan, alginic acid, polyphenols, plant sterols, lactic acid bacteria ・Hyperglyceridemia: tea catechins, globin peptides, isoprenoids ・Hyperglycemia, diabetes: wheat albumin, indigestible dextrin, tea polyphenols, isoprenoids, arteriosclerosis, cardiovascular diseases: carotenoids, polyphenols, n-3 fatty acids, obesity: tea catechins, diacylglycerols, cancer: carotenoids, flavonoids, Isoflavones, isothiocyanates, lactic acid bacteria, n-3 fatty acids, glycosphingolipids ・Osteoporosis: milk basic protein, isoflavones, lactic acid bacteria, oligosaccharides, menaquinone ・Tooth decay: xylitol, calcium monohydrogen phosphate ・Constipation: indigestion Indigestible dextrin, oligosaccharide ・Diarrhea: indigestible dextrin, oligosaccharide ・Inflammatory enteritis: amino acid, polyphenol, lactic acid bacteria ・Liver damage: isoprenoid, amino acid ・Gastritis: milk protein ・Gastrointestinal infection: peptide, antibody protein, lactic acid bacteria ・Asthma/atopy: polyphenols, lactic acid bacteria, oligosaccharides, n-3 fatty acids, glycosphingolipids/hay fever: polyphenols, lactic acid bacteria, oligosaccharides/dementia: lecithin, n-6 fatty acids, n-3 fatty acids/eyes Fatigue: Astaxanthin, Anthocyanin Insomnia: Glycine

(Classification based on acting fields)
Classification based on the place of action (classification by biological system) was proposed when the concept of the tertiary function of food was proposed, and is classified into immune system regulators, secretory system regulators, and nervous system regulators. It is a classification that divides into six types of factors, circulatory system regulatory factors, digestive system regulatory factors, and cell system regulatory factors.

(Classification based on mechanism of action)
Classification based on the mechanism of action includes induction of changes in intestinal flora, regulation of material transfer rate in the intestinal tract, binding and adsorption of nutrients in the intestinal tract, antibacterial action, inhibition of enzymes, and activity of enzymes and proteins. Classification based on mechanism of action, such as catabolism, antioxidant activity, and modification of hormone action.

<Food factor function>
A food factor function is a function that a food factor exhibits in a living body. Various food factor functions have been found for each food factor. -Aminobutyric acid (GABA) is known to lower blood pressure, and synephrine is known to have an adrenaline-like action.

<Food factor sensing-related gene (food factor sensing-related gene)>
A food factor sensing-related gene (food factor sensing-related gene) is a living organism-side gene responsible for the functional expression of a food factor. Food factor sensing-related genes are genes involved in the sensing system of food factors in vivo, and various genes are known for each food factor. Examples of food factor sensing-related genes, along with their food factors and characteristics, are shown in Tables 1-4 below.

<Euglena>
In the present embodiment, "euglena" taxonomically includes microorganisms classified into the genus Euglena, variants thereof, variants thereof, and closely related species of the family Euglenaceae.
Here, the genus Euglena is a group of eukaryotic organisms belonging to Excoverta, Euglenozoa, Euglenaphycea, Euglenoides, and Euglenaceae.

Species included in the genus Euglena specifically include Euglena chadefaudii, Euglena deses, Euglena gracilis, Euglena granulata, Euglena mutabilis, Euglena proxima, Euglena spirogyra, and Euglena viridis.
As Euglena, Euglena gracilis (E. gracilis), in particular, Euglena gracilis (E. gracilis) Z strain can be used, but in addition, Euglena gracilis (E. gracilis) Z strain mutant strain SM-ZK strain (chloroplast-deficient strain), variants of E. gracilis var. bacillaris, gene mutant strains such as chloroplast mutant strains of these species, and other Euglenoids such as Astaia longa.

Euglenoids are widely distributed in freshwater such as ponds and marshes, and may be used by separating from them, or any already isolated Euglenoids may be used.
The genus Euglena includes all its variants. These mutant strains also include those obtained by genetic methods such as recombination, transduction, and transformation.

In the culture of Euglena cells, the culture medium includes, for example, a culture medium supplemented with nutrients such as a nitrogen source, a phosphorus source, and minerals, such as a modified Cramer-Myers medium ((NH ₄ ) ₂ HPO ₄ 1.0 g/L , _KH2PO4 1.0 g/L, _MgSO4.7H2O 0.2 g/l, _{CaCl2.2H2O 0.02} g/l _{, Fe2 ( SO2} ) _{3.7H2O 3} mg/l, _MnCl2.4H2O 1.8 mg/l, _CoSO4.7H2O 1.5 mg/ _l , _{ZnSO4.7H2O 0.4} mg/ _l , _{Na2MoO4.2H2O 0.2 mg} /l, CuSO ₄ .5H ₂ O 0.02 g/l, thiamine hydrochloride (vitamin B1) 0.1 mg/l, cyanocobalamin (vitamin B12), (pH 3.5)) can be used. (NH ₄ ) ₂ HPO ₄ can also be converted to (NH ₄ ) ₂ SO ₄ or NH ₃ aq. In addition, known Hutner medium and Koren-Hutner medium prepared according to the description of Euglena physiology and biochemistry (edited by Shozaburo Kitaoka, Gakkai Shuppan Center Co., Ltd.) may also be used.

The pH of the culture medium is preferably 2 or more, and the upper limit is preferably 6 or less, more preferably 4.5 or less. By making the pH on the acidic side, photosynthetic microorganisms can grow more dominantly than other microorganisms, so that contamination can be suppressed.
Cultivation of Euglena cells may be performed by an open pond method in which sunlight is directly used, a light collection method in which sunlight collected by a light collecting device is sent through an optical fiber or the like, irradiated in a culture tank and used for photosynthesis, or the like.
In addition, Euglena cells can be cultured, for example, using a feeding batch method, but flask culture, culture using a fermenter, batch culture method, semi-batch culture method (fed-batch culture method), continuous culture method (perfusion culture method) or any other liquid culture method.
Separation of Euglena cells is carried out, for example, by centrifugation, filtration or simple sedimentation of the culture medium.

(Euglenoid algae)
In the present embodiment, Euglena living cells separated by centrifugation, filtration, sedimentation, or the like can be used as they are as the Euglenoid algae. Euglena living cells can be used as they are after being harvested from the culture vessel, but they are preferably washed with water or physiological saline. Alternatively, the Euglenoid alga body may be used in the form of a dispersion in a liquid such as water. In the present embodiment, it is preferable to use dry Euglena alga bodies obtained by freeze-drying or spray-drying Euglena living cells as Euglena alga bodies.
Furthermore, a mechanically processed product of alga bodies obtained by subjecting living Euglena cells to ultrasonic irradiation treatment or mechanical treatment such as homogenization may be used as the Euglena alga bodies. Alternatively, a dried mechanically processed product obtained by subjecting a mechanically processed product to a drying treatment may be used as the Euglenoid algal body.

(Euglena aqueous solvent extract)
In the present embodiment, "Euglena aqueous solvent extract" means an extract extracted from Euglena using an aqueous solvent, in particular, using water as an aqueous solvent, at 5 ° C. to 600 ° C., several seconds to several It is preferable to use a water extract or a hot water extract of Euglena extracted for 10 hours.
The water used for extraction does not necessarily have to be distilled, pure, or ultrapure water, and may, for example, be tap water or contain impurities, but may contain components that interfere with the extraction of the active ingredient. no water is preferred.

In the present embodiment, "water extract" means an extract with water at 0 to 50°C (excluding 0°C).
Here, "water" means water at 0 to 50°C (excluding 0°C).
The temperature of the water is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient. Especially preferred is 10 to 30°C.

In this embodiment, "hot water extract" means an extract with water at a temperature higher than 50°C, and can also be called a "hot water extract".
Here, “hot water” means water having a temperature higher than 50° C., and is a concept that includes “hot water”, including water in a boiling state. Moreover, it is not limited to hot water in a liquid state, but also includes hot water in a gaseous state and a supercritical state.
The temperature of the hot water is not particularly limited as long as the active ingredient can be extracted sufficiently without affecting the active ingredient. °C and 100°C or less.

The pH of the water used for extraction is not particularly limited as long as the active ingredient can be sufficiently extracted without affecting the active ingredient, preferably pH 4 to 10, more preferably pH 5 to 9. , particularly preferably pH 6-8.

In the present embodiment, water is used alone as the aqueous solvent, but the active ingredient can be sufficiently extracted without affecting the active ingredient, and usually one solvent that can be used for extraction is used. Alternatively, two or more kinds may be selected and used. Examples include, but are not limited to, water, alcohols, glycols, and the like. Alcohols include ethanol, methanol, n-propanol, isopropanol and the like. Glycols include butylene glycol and propylene glycol. Other aqueous solvents include acetone and the like. These solvents may be used alone or as an aqueous solution, or may be used as a mixed solvent of any two or three or more.

The temperature of the aqueous solvent used for extraction is, for example, 0° C. or higher, and is not particularly limited as long as it does not affect the active ingredients. Aqueous solvents in a boiling or supercritical state can also be used, but it is preferred to use aqueous solvents at 5°C to 600°C, more preferably 10°C to 200°C.
Therefore, the aqueous solvent for extraction includes an aqueous solvent in a boiling state or a supercritical state. The amount of the aqueous solvent used for extraction is preferably an amount that can sufficiently dissolve the water-soluble active ingredient contained in Euglena.

The extraction method is also not particularly limited, and for example, extraction can be performed by the method shown below, but the extraction method is not limited to this, and can be freely selected and used. For example, a dry powder of Euglena algae is immersed in an aqueous solvent for a predetermined time and then centrifuged or filtered, or a dry powder of Euglena algae is added to an aqueous solvent and shaken to uniformly disperse, followed by centrifugation or filtration. method, etc.
Moreover, in order to promote extraction, it is also possible to heat the aqueous solvent after adding Euglena.

Euglena can be extracted with water by a conventional method as shown below, but not limited thereto. For example, the Euglena tissue and water are placed in a container and allowed to stand for a predetermined period of time with appropriate stirring or shaking, and the obtained extract can be used as a water extract as it is. Moreover, for example, the supernatant obtained by centrifuging such an extract can also be used as an aqueous extract. Also, such an extract or supernatant can be concentrated and dried to remove water and used as an aqueous extract. The water extraction may be carried out by adding a small amount of alcohol, preferably ethanol, to water in an amount of, for example, 10% by mass or less, in order to increase the extraction efficiency and shorten the extraction time.
The extraction time for water extraction is not particularly limited as long as the active ingredient is extracted, and can be appropriately set in the range of several seconds to several tens of hours depending on the extraction temperature.

Extraction with hot water can be performed by a commonly used method as shown below, but is not limited thereto. After Euglena is introduced into a commonly used extractor together with water, it is extracted by heating. When boiling water or water in a supercritical state is used for extraction, it is necessary to use an extractor that can withstand the vapor pressure of water. The pressure during extraction can be set to 1 to 5000 atmospheres, preferably 60 to 400 atmospheres.
When the extraction is performed under high temperature and high pressure, if the extraction time is too long, the active ingredients may decompose or chemical reactions may occur. Therefore, when the extraction is performed under high temperature and high pressure, the extraction time is short, for example, preferably within 3 minutes, more preferably within 1 minute, and particularly preferably within 30 seconds.

The extracted Euglena extract can be used as it is as an active ingredient such as the food factor sensing-related gene expression promoter according to the present embodiment. Filtration, silica gel chromatography, reverse-phase or normal-phase high-performance liquid chromatography, etc.) can be used to fractionate and use highly active fractions.
Alternatively, the Euglena extract or a fraction thereof can be concentrated and dried to remove the aqueous solvent, and this can be used as an aqueous solvent extract.

(paramylon)
"Paramylon" is a porous polymer (β-1,3-glucan) in which about 700 glucoses are polymerized by β-1,3-bonds, and is a storage polysaccharide contained in the genus Euglena. is. Paramylon particles are flattened spheroidal particles and are formed by helically intertwined β-1,3-glucan chains.

Paramylon exists as granules in Euglena cells of all species and varieties, and the number, shape, and uniformity of the particles are characteristic depending on the species.
Paramylon is composed only of glucose, and the average degree of polymerization of paramylon obtained from E. gracilis Z wild strain and chloroplast-deficient strain SM-ZK is about 700 in terms of glucose units.
Paramylon is insoluble in water and hot water, but soluble in dilute alkali, strong acid, dimethyl sulfoxide, formaldehyde and formic acid.
The average density of paramylon is 1.53 for E. gracilis Z and 1.63 for E. gracilis var. bacillaris SM-L1.
According to X-ray analysis using the powder pattern method, paramylon has a gentle helical structure in which three straight-chain β-1,3-glucans are twisted like a right-handed rope. Several of these glucan molecules aggregate to form paramylon granules. Paramylon granules have a very large crystal structure portion, accounting for about 90%, and are compounds with the highest crystal structure ratio among polysaccharides (Euglena, Physiology and Biochemistry, edited by Shozaburo Kitaoka, Gakkai Shuppan Center).
The particle size distribution of paramylon (manufactured by Euglena Co., Ltd.) has a median diameter of 1.5 to 2.5 μm when measured with a laser diffraction/scattering particle size distribution analyzer.

Paramylon particles are isolated from cultured Euglena cells by any appropriate method and purified into fine particles, and are usually provided as powder.
For example, paramylon particles can be (1) cultured Euglena cells in any appropriate medium, (2) separation of Euglena cells from the medium, (3) isolation of paramylon from the separated Euglena cells, ( 4) purification of the isolated paramylon, and optionally (5) cooling and subsequent lyophilization.
Isolation of paramylon is performed, for example, using non-ionic or anionic detergents of the type that are largely biodegradable. Purification of paramylon occurs substantially simultaneously with isolation.

Isolation and purification of paramylon from Euglena are well known, for example, E. Ziegler, "Die naturlichen und kunstlichen Aromen" Heidelberg, Germany, 1982, Chapter 4.3 "Gefriertrocken", DE4328329, or JP2003-529538 described in the official gazette.

(Processed product of paramylon)
Processed products of paramylon include water-soluble paramylon obtained by chemically or physically treating paramylon by various known methods, sulfated paramylon, and paramylon derivatives.

Examples of processed products of paramylon include amorphous paramylon.
Amorphous paramylon is a substance obtained by amorphizing crystalline paramylon derived from Euglena.
Amorphous paramylon has a relative crystallinity of 1 to 20% with respect to crystalline paramylon produced from Euglena by a known method.
However, this relative crystallinity is obtained by the method described in JP-A-2011-184592.
That is, amorphous paramylon and paramylon were each pulverized with a pulverizer (ball mill MM400 manufactured by Retsh) for 5 minutes at a frequency of 20 times/second, and then analyzed using an X-ray diffractometer (H'PertPRO manufactured by Spectris). At a voltage of 45 KV and a tube current of 40 mA, scanning is performed in the range of 2θ from 5° to 30° to obtain diffraction peaks Pc and Pa near 2θ=20° of paramylon and amorphous paramylon.
Using the values of Pc and Pa, the relative crystallinity of amorphous paramylon is
Relative crystallinity of amorphous paramylon = Pa/Pc x 100 (%)
Calculated by

Amorphous paramylon is prepared according to the method described in JP-A-2011-184592 by subjecting crystalline paramylon powder to alkali treatment, followed by neutralization with acid, followed by washing, water removal, and drying.
Processed products of paramylon also include water-soluble paramylon obtained by chemically or physically treating paramylon by various known methods, sulfated paramylon, and paramylon derivatives.

<Food factor sensing-related gene expression promoter>
The food factor sensing-related gene expression promoter according to the present embodiment is a food factor sensing-related gene expression promoter (food factor sensing-related gene expression promoter) containing an Euglena-derived substance as an active ingredient.
“Euglena-derived substances” include Euglena algae bodies such as Euglena living cells and dried Euglena algae bodies, Euglena aqueous solvent extracts, processed products of Euglena algae bodies, paramylon, processed products of paramylon, and the like.

“Food factor sensing-related gene expression promoter” refers to an agent that promotes expression of a food factor sensing-related gene, and is synonymous with “food factor sensing-related gene expression level increasing agent”. An increase in gene amount can be evaluated by measuring the expression level of the gene by a known method such as the PCR method.

The food factor sensing-related gene expression promoter of the present embodiment promotes the expression of food factor sensing-related genes involved in sensing food factors, resulting in proteins related to food factor sensing (food factor reception body) expression.

<Food factor function enhancer>
The food factor sensing-related gene expression promoter according to the present embodiment contains an Euglena-derived substance as an active ingredient, and can also be used as a food factor function enhancer for enhancing food factor function.

“Enhancing the function of food factors” refers to enhancing the functions of food factors in vivo compared to the state before ingestion of the Euglena-derived substance, which is the active ingredient of the food factor function enhancer of the present embodiment.

The food factor sensing-related gene expression promoter of the present embodiment promotes the expression of food factor sensing-related genes in vivo and promotes the expression of proteins (food factor receptors) related to food factor sensing. It has the effect of enhancing the function of food factors.

<Food additives for enhancing food factor functions>
The food factor function enhancer of the present embodiment contains a Euglena-derived substance as an active ingredient, and is also used as a food additive for enhancing the function of a food factor (food additive for enhancing the function of a food factor) for enhancing the function of a food factor. be able to.
Euglena-derived substances can enhance the function of food factors, so by adding them to foods such as foods for specified health uses, foods with nutrient function claims, foods with function claims, foods for hospital patients, supplements, etc. The food factor function of the included food factor can be enhanced.

<Application>
The food factor sensing-related gene expression promoter and food factor function enhancer containing the Euglena-derived substance according to the present embodiment as an active ingredient are configured as food compositions such as health foods and pharmaceutical compositions, and have a sensing function of food factors. is administered to a living body in which the

Euglenoid-derived substances such as Euglenoid algae and paramylon can be ingested as food and have no side effects, so they can be administered even before a specific disease is definitively diagnosed.

The food factor sensing-related gene expression promoter and food factor function enhancer containing the Euglena-derived substance of the present embodiment as an active ingredient are used in food due to some reason such as lifestyle, aging, diet, lack of exercise, etc. It can be administered continuously for a long period of time to people who are likely to have decreased factor-sensing function.

Subjects to whom the food factor sensing-related gene expression promoter and food factor function enhancer containing the Euglena-derived substance as an active ingredient according to the present embodiment are administered are not limited to those with the above conditions and animals other than humans. do not have.

In addition, it is possible to administer food factor sensing-related gene expression promoters and food factor function enhancers containing Euglena-derived substances as active ingredients to humans aged 40 and over, particularly elderly people aged 60 or 65 and over. can.
Humans over the age of 40, especially those over the age of 60 or 65, tend to lose their ability to sense food factors as they age. The function-enhancing action of the factor can improve the sensing function of the food factor.

According to the food factor sensing-related gene expression promoter of the present embodiment, it is possible to improve the sensing function of a food factor in a living body. It is expected to be applied to improve the function of food factors in food.

In addition, the food factor sensing-related gene expression promoter containing the Euglena-derived substance according to the present embodiment as an active ingredient is added with a pharmacologically acceptable additive, and is used in compositions such as pharmaceutical compositions and food compositions. can be used as

(Food composition)
The Euglena-derived substance of the present embodiment can also be used in foods.
In the field of food, the food composition for promoting food factor sensing-related gene expression of the present embodiment is an effective amount of Euglena-derived substance that can effectively exhibit the food factor sensing-related gene expression promoting action. By blending, it is possible to provide a food composition having the action.
It is also possible to provide food compositions by combining Euglena-derived substances with foods for specified health uses, foods with nutrient function claims, foods with function claims, foods for hospital patients, supplements, and the like.
Examples of the food composition include seasonings, processed meat products, processed agricultural products, beverages (soft drinks, alcoholic drinks, carbonated drinks, milk drinks, fruit juice drinks, tea, coffee, nutritional drinks, etc.), powdered drinks (powder juices, powdered soups, etc.), concentrated beverages, confectionery (candies, cookies, biscuits, gums, gummies, chocolates, etc.), breads, cereals and the like. In the case of food for specified health uses, food with nutrient function claims, food with function claims, etc., the shape may be capsule, troche, syrup, granule, powder, or the like.

Foods for Specified Health Uses are foods that contain functional health ingredients that affect physiological functions, etc., and can be labeled as suitable for specific health uses with the permission of the Director-General of the Consumer Affairs Agency. be.
A food with nutrient function claims is a food that is used to supplement nutritional ingredients (vitamins and minerals), and the functions of the nutritional ingredients are indicated. In order to be sold as a food with nutrient function claims, the amount of nutrients contained in the recommended daily intake must be within the specified upper and lower limits. also need to
Foods with function claims are foods labeled with functionalities based on scientific evidence under the responsibility of the business operator. Information on the grounds for safety and functionality was submitted to the Commissioner of the Consumer Affairs Agency before sales.

In the food composition according to the present embodiment, in addition to the Euglena-derived substance, it is possible to freely select and add one or more components that can be used in ordinary food compositions. For example, all additives that can be commonly used in the food field, such as various seasonings, preservatives, emulsifiers, stabilizers, fragrances, coloring agents, preservatives, and pH adjusters, can be included.

In addition to the Euglena-derived substance, one or more other substances known to have food factor sensing-related gene expression promoting effects can be added to the food composition according to the present embodiment.

(Pharmaceutical composition)
The food factor sensing-related gene expression promoter containing the Euglena-derived substance according to the present embodiment as an active ingredient can be used as a pharmaceutical composition.
In the field of medicine, the food factor sensing-related gene expression promoter of the present embodiment contains a pharmaceutically acceptable carrier and additives together with an amount of Euglena-derived substances that can effectively exhibit the food factor sensing-related gene expression promoting action. By blending, a pharmaceutical composition having the action is provided. The pharmaceutical composition may be a drug or a quasi-drug.
The pharmaceutical composition may be applied internally or topically. Therefore, the pharmaceutical composition is used in the form of an oral preparation, an injection such as an intravenous injection, a subcutaneous injection, an intradermal injection, an intramuscular injection and/or an intraperitoneal injection, a transmucosal application, a transdermal application, and the like. be able to.
The dosage form of the pharmaceutical composition can be appropriately set depending on the application form. Semi-solid formulations such as ointments or gels are included.

The pharmaceutical composition according to this embodiment can contain one or more pharmaceutically acceptable additives by freely selecting them.
For example, when the pharmaceutical composition according to this embodiment is applied to an oral formulation, for example, excipients, binders, disintegrants, surfactants, preservatives, coloring agents, flavoring agents, fragrances, stabilizers, preservatives All additives that can be commonly used in the field of pharmaceutical formulations, such as agents, antioxidants, etc., can be contained. In addition, a drug delivery system (DDS) can be used to prepare a sustained-release preparation or the like.

In addition to the Euglena-derived substance, one or more other substances known to have food factor sensing-related gene expression promoting effects can be added to the pharmaceutical composition according to the present embodiment.

<Dosage and dosage>
The food factor sensing-related gene expression promoter of the present embodiment may be used, for example, by oral administration in the form of powders, capsules, tablets, granules, liquids, syrups, or the like.
The dosage and dosage form of the food factor sensing-related gene expression promoter of the present embodiment may be appropriately selected depending on the subject, disease state and progress thereof, and other conditions. For example, when Euglena is selected as the Euglena-derived substance and orally administered to humans (adults) for the purpose of obtaining the food factor sensing-related gene expression promoting effect, Euglena is generally added to 1 dry weight per day. It is recommended to administer continuously at a rate of 1 to 2 times a day (morning and evening) and at least 5 times a week so that the dose is about 5000 mg, preferably 100 to 3000 mg, more preferably 500 to 2000 mg. .

EXAMPLES The present invention will be specifically described below based on specific examples, but the present invention is not limited to these.
In the following tests, Euglena powder (Example 1) and Paramylon powder (Example 2) were used as Euglena-derived substances to clarify the effects of continuous intake of Euglena or Paramylon on the expression of genes related to food factor sensing. purpose.

<Example 1>
Euglena gracilis powder (Euglena algae, manufactured by Euglena Co., Ltd.), which is a substance derived from Euglena, was used as a food factor sensing-related gene expression promoter.

<Example 2>
Paramylon powder (manufactured by Euglena Co., Ltd.), which is a substance derived from Euglena, was used as a gene expression promoter related to food factor sensing.

<Test 1 Examination of the effect of continuous intake of Euglena-derived substances on the expression level of food factor sensing-related genes>
(Test method)
・Feed preparation CE-2 meal (purchased from CLEA Japan) was mixed to contain 2% Euglena powder to prepare a Euglena meal. Similarly, the CE-2 meal was mixed with 2% paramylon powder to prepare a paramylon meal. The nutritional composition of the CE-2 diet is shown in Tables 5-7.

Feeding test 12-week-old C57BL / 6J male mice were divided into 3 groups so that the average body weight was equal, and each was a control group (C), a Euglena group (E), and a paramylon group (P) (n = 6). The control group was fed CE-2 (regular diet), the Euglena group was fed CE-2 containing 2% euglena (euglena diet), and the paramylon group was fed CE-2 containing 2% paramylon (paramylon diet) (Table 8). After a 4-week feeding period, quadriceps muscles and livers were harvested from the mice.

・Gene expression analysis After extracting RNA from the collected organs using Tri reagent (Cosmo Bio), a mixture of RNA from 6 animals in each group (n = 1) was used as an analysis sample, and a microarray (Agilent SurePrint G3 Mouse Gene expression was measured using the GE microarray kit 8x60K v2).

(Results of Test 1)
Results in liver are shown in FIGS. 1A-1D and FIG. 2, and results in quadriceps are shown in FIGS. 3A-3C and FIG.
In the liver liver, Adora2a (SEQ ID NO: 1), Adora3 (SEQ ID NO: 2), Adra1a (SEQ ID NO: 3), Bcl2 (SEQ ID NO: 4), Ccr2 (SEQ ID NO: 5), Ccr5 (SEQ ID NO: 6), Ccr6 (SEQ ID NO: 7), Cd36 (SEQ ID NO: 8), Ffar1 (SEQ ID NO: 9), Gabbr1 (SEQ ID NO: 10), Glra4 (SEQ ID NO: 11), Hic2 (SEQ ID NO: 12), IL1a (SEQ ID NO: 13), IL1b (SEQ ID NO: 14), IL6 (SEQ ID NO: 15), IL16 (SEQ ID NO: 16), Ppara (SEQ ID NO: 17), Tlr1 (SEQ ID NO: 18), Tlr3 (SEQ ID NO: 19), Tlr5 (SEQ ID NO: 20), Tlr6 ( 21 genes of SEQ ID NO: 21) were increased 1.5 times or more compared to the control group.
In the liver, the paramylon group includes Adora3 (SEQ ID NO: 2), Adra1a (SEQ ID NO: 3), Ager (SEQ ID NO: 22), Bcl2 (SEQ ID NO: 4), Ccr5 (SEQ ID NO: 6), Cd4 (SEQ ID NO: 23), 13 of Esrrb (SEQ ID NO: 24), IL1b (SEQ ID NO: 14), IL1r1 (SEQ ID NO: 25), Ltb4r1 (SEQ ID NO: 26), Tlr6 (SEQ ID NO: 21), Olr1 (SEQ ID NO: 27), Stx3 (SEQ ID NO: 28) Genes were increased more than 1.5-fold compared to the control group.
In the liver, six genes Adra1a (SEQ ID NO: 3), Adora3 (SEQ ID NO: 2), Bcl2 (SEQ ID NO: 4), Ccr5 (SEQ ID NO: 6), IL1b (SEQ ID NO: 14), Tlr6 (SEQ ID NO: 21) , both the euglena group and the paramylon group increased more than 1.5 times compared to the control group.

Therefore, in the liver, euglena and paramylon as Euglena-derived substances have the effect of increasing food factor sensing-related genes. It turned out that it can be used as an object.

・Quadriceps muscle In the quadriceps muscle, in the Euglena group, Bcl2l1 (SEQ ID NO: 29), Ccr6 (SEQ ID NO: 7), Esr1 (SEQ ID NO: 30), Esrrb (SEQ ID NO: 24), Ffar3 (SEQ ID NO: 31), Glrb (SEQ ID NO: 32), Ppara (SEQ ID NO: 17), Rxra (SEQ ID NO: 33), IL6ra (SEQ ID NO: 34), Tlr6 (SEQ ID NO: 21), Vdr (SEQ ID NO: 35) compared with the control group It increased more than 1.5 times.
In addition, in the quadriceps muscle, in the paramylon group, Bcl2l1 (SEQ ID NO: 29), Gabbr1 (SEQ ID NO: 10), Gpr84 (SEQ ID NO: 36), IL6ra (SEQ ID NO: 34 ), IL16 (SEQ ID NO: 16), Sirt1 (sequence No. 37) and Tlr4 (SEQ ID NO: 38) increased 1.5 times or more compared to the control group.
In the quadriceps femoris muscle, Bcl2l1 (SEQ ID NO: 29) and IL6ra (SEQ ID NO: 3 4 ) genes increased more than 1.5 times in both the euglena group and the paramylon group compared to the control group. was

Therefore, in the quadriceps femoris, Euglena and paramylon as Euglena-derived substances have the effect of increasing food factor sensing-related genes. It was found that it can be used as a food composition for food.

(Food Factor Function Enhancement Effect of Euglena-Derived Substance)
Since the Euglena-derived substance promotes the expression of genes related to food factor sensing, it is possible to enhance the function of food factors, as shown in FIGS.
Specifically, as shown in FIG. 5, by ingesting Euglena, the expression of Ppara (SEQ ID NO: 17), Vdr (SEQ ID NO: 35), and Ffar1 (SEQ ID NO: 9), which are genes related to food factor sensing, is increased. Therefore, the lipid metabolism improving action, which is the food factor function of unsaturated fatty acids, the bone protective action, which is the food factor function of vitamin D, and the antidiabetic action, which is the food factor function of long-chain fatty acids, are enhanced. is assumed.

In addition, as shown in FIG. 6, ingestion of paramylon promotes the expression of Sirt1 (SEQ ID NO: 37), Gabbr1 (SEQ ID NO: 10), and Adra1a (SEQ ID NO: 3), which are genes related to food factor sensing. Therefore, it is assumed that the anti-aging effect, which is a food factor function of flavonoids, the blood pressure-lowering effect, which is a food factor function of GABA, and the adrenaline-like effect, which is a food factor function of synephrine, are enhanced.

<Test 2 Examination of the effect of continuous intake of Euglena-derived substances on the expression of soy isoflavone sensor-related genes>
(Test method)
・Feed preparation CE-2 meal (purchased from CLEA Japan) was mixed to contain 2% Euglena powder to prepare a Euglena meal.

Feeding test 12-week-old C57BL / 6J male mice were divided into 3 groups so that the average body weight was equal, and each was a control group (C), a Euglena group (E), and a paramylon group (P) (n = 6). The control group was fed CE-2 (regular diet), and the Euglena group was fed CE-2 containing 2% Euglena (Euglena diet). After a 4-week feeding period, quadriceps were harvested from the mice.

- Gene expression analysis Total RNA was recovered from the quadriceps muscle, cDNA was synthesized, and the expression level of each gene was measured by real-time PCR. The expression level of each gene was corrected using β-actin as an internal standard gene.

(Results of test 2)
FIG. 7 shows the measurement results of the expression levels of Esr1 (SEQ ID NO: 30) and Esrrb (SEQ ID NO: 24) in the quadriceps muscle, and FIG. 8 shows the measurement results of the Ctsd (SEQ ID NO: 39) expression level.

As a result of measuring the expression levels of genes that fluctuated in the quadriceps femoris muscle by real-time PCR, an increase in the expression of Esr1 (SEQ ID NO: 30), known as a soy isoflavone sensor, and its related gene Esrrb (SEQ ID NO: 24) was confirmed ( Figure 7). In addition, the expression level of Ctsd (SEQ ID NO: 39), whose expression is induced by Esr1 (SEQ ID NO: 30) activation, was measured by real-time PCR. confirmed (Fig. 8).

Therefore, by ingesting Euglena, the expression of Esr1 (SEQ ID NO: 30), Esrrb (SEQ ID NO: 24), and Ctsd (SEQ ID NO: 39), which are genes related to food factor sensing, can be promoted. It is assumed that factor function is enhanced.

Claims (12)

containing Euglena as an active ingredient,
promoting the expression of Ppara (SEQ ID NO: 17);
A food composition for promoting food factor sensing-related gene expression, which is used for enhancing the lipid metabolism improving action of unsaturated fatty acids, which is the food factor function. containing Euglena as an active ingredient,
used to promote the expression of Vdr ( SEQ ID NO: 35),
A food composition for promoting gene expression related to food factor sensing, characterized by being used for enhancing the bone-protecting action, which is the food factor function of vitamin D. containing Euglena as an active ingredient,
used to promote expression of Ffar1 (SEQ ID NO: 9),
A food composition for promoting gene expression related to food factor sensing, which is used to enhance the antidiabetic action of long-chain fatty acids, which is the food factor function. Contains paramylon as an active ingredient,
used to promote the expression of Sirt1 (SEQ ID NO: 37),
A food composition for promoting gene expression related to food factor sensing, characterized by being used for enhancing the anti-aging effect, which is the food factor function of flavonoids. Contains paramylon as an active ingredient,
used to promote expression of Gabbr1 (SEQ ID NO: 10),
A food composition for promoting food factor sensing-related gene expression, which is used for enhancing the blood pressure-lowering action of GABA, which is a food factor function. Contains paramylon as an active ingredient,
used to promote the expression of Adra1a (SEQ ID NO: 3),
A food composition for promoting gene expression related to food factor sensing, characterized by being used for enhancing adrenaline-like action, which is a food factor function of synephrine. containing Euglena as an active ingredient,
promoting the expression of Ppara (SEQ ID NO: 17);
A food factor sensing-related gene expression promoter, which is used to enhance the lipid metabolism-improving action of unsaturated fatty acids, which is the function of food factors. containing Euglena as an active ingredient,
used to promote the expression of Vdr ( SEQ ID NO: 35),
A food factor sensing-related gene expression promoter, characterized by being used for enhancing the bone protective action, which is the food factor function of vitamin D. containing Euglena as an active ingredient,
used to promote expression of Ffar1 (SEQ ID NO: 9),
A food factor-sensing-related gene expression promoter characterized by being used for enhancing the antidiabetic action, which is the food factor function of long-chain fatty acids. Contains paramylon as an active ingredient,
used to promote the expression of Sirt1 (SEQ ID NO: 37),
A food factor sensing-related gene expression promoter characterized by being used for enhancing the anti-aging effect, which is the food factor function of flavonoids. Contains paramylon as an active ingredient,
used to promote expression of Gabbr1 (SEQ ID NO: 10),
A food factor sensing-related gene expression promoter, which is used for enhancing the blood pressure-lowering action of GABA, which is a food factor function. Contains paramylon as an active ingredient,
used to promote the expression of Adra1a (SEQ ID NO: 3),
A food factor sensing-related gene expression promoter, which is used to enhance the adrenaline-like action, which is the food factor function of synephrine.

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