The composition of vascular flexibility upward

Abstract

An object of the present invention is to provide a composition having excellent effects of improving vascular flexibility and vascular endothelial function. [Solution] A composition having the effect of improving vascular flexibility and vascular endothelial function, characterized by containing pine bark and at least one selected from the group consisting of ginger, elastin, and vitamin E.

Description

[Technical Field]

[0001] The present invention relates to a composition for improving vascular flexibility and a composition for improving vascular endothelial function, each of which contains pine bark and at least one member selected from the group consisting of ginger, elastin, and vitamin E. The present invention also relates to an oral composition containing pine bark, ginger, and elastin. [Background technology]

[0002] In recent years, lifestyle-related diseases have been increasing due to changes in diet and lifestyle. Lifestyle-related diseases are a general term that includes not only hypertension, hyperlipidemia, and diabetes, but also angina pectoris, myocardial infarction, and cerebral circulatory disorders, the onset of which is thought to be preventable by improving lifestyle habits. It has been revealed that reduced vascular flexibility and reduced vascular endothelial function are involved as factors in these diseases. Furthermore, because lifestyle-related diseases are chronic, their prevention and treatment require a long period of time. Therefore, there is a need for the development of foods that can safely and effectively improve vascular flexibility and vascular endothelial function over the long term. Known food ingredients with such effects include hops and chlorogenic acid (Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Patent Publication No. 2005-104951 [Patent Document 2] Patent Publication No. 2003-261444 Summary of the Invention [Problem to be solved by the invention]

[0004] However, according to the research conducted by the present inventors, the effects of hops and chlorogenic acid on improving vascular flexibility and vascular endothelial function are not sufficient.

[0005] Therefore, the present inventors have conducted various studies with the objective of providing a composition that contains ingredients that can be safely ingested over a long period of time and that exhibits excellent effects of improving vascular flexibility and vascular endothelial function.

[0006] The present inventors have conducted extensive research to solve the above problems and have surprisingly found that a combination of pine bark with at least one selected from the group consisting of ginger, elastin, and vitamin E exhibits excellent effects of improving vascular flexibility and vascular endothelial function. The present invention was completed based on this finding.

[0007] The outline of the present invention is as follows. [1] A composition for improving vascular flexibility, comprising pine bark and at least one selected from the group consisting of ginger, elastin, and vitamin E. [2] A composition for improving vascular endothelial function, comprising pine bark and at least one selected from the group consisting of ginger, elastin, and vitamin E. [3] An oral composition comprising pine bark, ginger, and elastin. [4] An oral composition comprising pine bark, ginger, elastin, and vitamin E. [5] A composition for improving vascular flexibility, comprising pine bark and at least two or more members selected from the group consisting of ginger, elastin, and vitamin E. [6] A composition for improving vascular endothelial function, comprising pine bark and at least two or more members selected from the group consisting of ginger, elastin, and vitamin E. [7] A composition for improving vascular flexibility, comprising pine bark, ginger, elastin, and vitamin E. [8] A composition for improving vascular endothelial function, comprising pine bark, ginger, elastin, and vitamin E.

[0008] According to the present invention, by containing at least one selected from the group consisting of pine bark, ginger, elastin, and vitamin E, a composition can be provided which has excellent effects of improving vascular flexibility and vascular endothelial function. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 is a diagram showing the expression levels of NO synthase genes in Examples and Comparative Examples. DETAILED DESCRIPTION OF THE INVENTION

[0010] The present invention will be described in detail below, but the present invention is not limited to the following embodiments.

[0011] The composition of the present invention is characterized by containing, together with pine bark, at least one component selected from ginger, elastin, and vitamin E. The component selected from ginger, elastin, and vitamin E to be used together with pine bark may be used alone or in combination of two or three components, but from the viewpoint of improving vascular flexibility and endothelial function, it is preferable to use two or three components in combination, and it is particularly preferable to use three components in combination.

[0012] Each component contained in the composition of the present invention will be described below. [pine bark] The present invention is characterized by the use of pine bark. Examples of pine species from which the pine bark used in the present invention can be obtained include, but are not limited to, French maritime pine (Pinus Martima), larch, black pine, Japanese red pine, Pinus densiflora, Pinus sieboldii, Pinus sieboldii, Pinus sieboldii, Pinus palustris ... sieboldii, and Pinus sieboldii. Among these, French maritime pine is preferred in terms of its ability to improve vascular flexibility and vascular endothelial function.

[0013] In the present invention, pine bark can be processed and used. Examples of processed pine bark products include chips, crushed products, squeezed products, extracts, and dried powders thereof. Considering formulation feasibility, the pine bark used in the present invention is preferably crushed, squeezed, extract, or dried powder thereof, as these are easy to apply. From the viewpoint of improving vascular flexibility and vascular endothelial function, extracts or dried powders thereof are more preferred. The processed pine bark product may be produced by a method commonly known to those skilled in the art, or may be a product available on the market. For example, a pine bark extract manufactured by Toyo Shinyaku Co., Ltd. can be used.

[0014] Examples of extraction solvents used to obtain a pine bark extract include water, organic solvents, and aqueous organic solvents (e.g., aqueous alcohols such as aqueous ethanol). Examples of organic solvents include methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, acetone, hexane, cyclohexane, propylene glycol, aqueous ethanol, aqueous propylene glycol, ethyl methyl ketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oils and fats, 1,1,1,2-tetrafluoroethane, and 1,1,2-trichloroethene. These water and organic solvents may be used alone or in combination. Water is preferably used as the extraction solvent. The temperature of the solvent used during extraction is not limited as long as it is below the boiling point of the solvent.

[0015] The method for obtaining a pine bark extract is not particularly limited, and examples thereof include a heat extraction method, a supercritical fluid extraction method, a liquid carbon dioxide batch method, a liquid carbon dioxide reflux method, and a supercritical carbon dioxide reflux method. A combination of multiple extraction methods may also be used. By combining multiple extraction methods, it is possible to obtain pine bark extracts with various compositions.

[0016] Supercritical fluid extraction is a method of extraction using a supercritical fluid, which is a fluid in a state exceeding the gas-liquid critical point (critical temperature, critical pressure) of a substance. Examples of supercritical fluids that can be used include carbon dioxide, ethylene, propane, and nitrous oxide (laughing gas), with carbon dioxide being preferred.

[0017] The supercritical fluid extraction method includes an extraction step in which a target component is extracted with a supercritical fluid, and a separation step in which the target component is separated from the supercritical fluid. The separation step may involve extraction and separation by pressure change, extraction and separation by temperature change, or extraction and separation using an adsorbent or absorbent.

[0018] Alternatively, supercritical fluid extraction may be performed using the entrainer addition method. In this method, approximately 2-20 w / v% of an extracting fluid, such as ethanol, propanol, n-hexane, acetone, toluene, or other aliphatic lower alcohols, aliphatic hydrocarbons, aromatic hydrocarbons, or ketones, is added to the extracting fluid, and supercritical fluid extraction is performed using this fluid. This method dramatically increases the solubility of the target extract, such as oligomeric proanthocyanidins (OPCs) or catechins, in the extracting solvent or enhances the separation selectivity, resulting in an efficient method for obtaining pine bark extracts.

[0019] Supercritical fluid extraction has the advantages of being able to operate at relatively low temperatures, making it applicable to substances that change or decompose at high temperatures, leaving no residual extracted fluid, and simplifying the process by eliminating steps such as desolvation, since the solvent can be recycled.

[0020] From the viewpoint of safety, it is preferable to purify the pine bark extract obtained by the above extraction using a column method or a batch method. Examples of the column method include purification methods using an adsorptive carrier such as Diaion HP-20, Sephadex-LH20, or chitin.

[0021] The pine bark extract used in the present invention contains proanthocyanidins as one of its main components. Proanthocyanidins are a group of compounds consisting of condensation polymers with a degree of polymerization of 2 or more, whose constituent units are flavan-3-ol and / or flavan-3,4-diol.

[0022] The pine bark extract used in the present invention preferably contains, as proanthocyanidins, condensation polymers having a degree of polymerization of 2 or more. In particular, proanthocyanidins containing a large amount of condensation polymers having a low degree of polymerization are preferred. Examples of condensation polymers having a low degree of polymerization include condensation polymers having a degree of polymerization of 2 to 30 (dimers to tridecamers). From the viewpoint of improving vascular flexibility and vascular endothelial function, condensation polymers having a degree of polymerization of 2 to 10 (dimers to decamers) are preferred, and condensation polymers having a degree of polymerization of 2 to 4 (dimers to tetramers) are particularly preferred. In this specification, polymers having a degree of polymerization of 2 to 4 are referred to as OPCs (oligomeric proanthocyanidins).

[0023] The proportion of pine bark to be incorporated into the composition of the present invention is not particularly limited, and can be appropriately set within a wide range depending on various conditions such as the purpose, shape, and target of use. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the proportion is preferably 0.001 to 50% by mass, and from the viewpoint of improving vascular flexibility and vascular endothelial function, 0.01 to 40% by mass is more preferable, and 0.1 to 30% by mass is particularly preferable. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the proportion is preferably 0.0001 to 30% by mass, more preferably 0.001 to 20% by mass, and particularly preferably 0.01 to 10% by mass.

[0024] [Ginger] In the present invention, ginger can be used together with pine bark. The ginger used in the present invention is not particularly limited as long as it is ginger (Zingiber officinale), a perennial plant of the Zingiberaceae family. Specific ginger varieties include Sanshu ginger, Omi ginger, Yanaka ginger, Kintoki ginger, Shizuoka No. 4, yellow ginger, earth ginger, and Otafuku ginger. However, Kintoki ginger is preferred in terms of its effects of improving vascular flexibility and vascular endothelial function.

[0025] The form of ginger used in the present invention is not particularly limited, but the rhizome can be used as is, or ginger can be processed and used. Examples of processed ginger products include chips, pulverized products, squeezed products, extracts, and dried powders thereof. Considering formulation feasibility, the ginger used in the present invention is preferably pulverized, squeezed, extract, or dried powder thereof, as these are easy to apply. In terms of the effects of improving vascular flexibility and vascular endothelial function, pulverized products, extracts, or dried powders thereof are more preferred, with pulverized products or dried powders thereof being particularly preferred. The processed ginger product may be one produced by a method commonly known to those skilled in the art, or may be one that is commercially available.

[0026] Examples of extraction solvents used to obtain ginger extract include water, organic solvents, and aqueous organic solvents (e.g., aqueous alcohols such as aqueous ethanol). Examples of organic solvents include methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, butane, acetone, hexane, cyclohexane, propylene glycol, aqueous ethanol, aqueous propylene glycol, ethyl methyl ketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oils and fats, 1,1,1,2-tetrafluoroethane, and 1,1,2-trichloroethene. These water and organic solvents may be used alone or in combination. From the viewpoint of improving vascular flexibility and vascular endothelial function, water, ethanol, or aqueous ethanol is preferred as the extraction solvent, with water being particularly preferred. The temperature of the solvent used during extraction is not limited as long as it is below the boiling point of the solvent.

[0027] The proportion of ginger to be incorporated into the composition of the present invention is not particularly limited, and can be appropriately set within a wide range depending on various conditions such as the purpose, shape, and subject of use. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the proportion is preferably 0.0001 to 20% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, 0.001 to 10% by mass is more preferable, and 0.01 to 5% by mass is particularly preferable. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the proportion is preferably 0.00001 to 10% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, 0.0001 to 5% by mass is more preferable, and 0.001 to 1% by mass is particularly preferable.

[0028] [Elastin] The present invention can use elastin together with pine bark. Elastin is a major elastic protein present, along with collagen, in mammals, birds, fish, etc., particularly in the aorta, ligamentum nuchale, ligamentum flavum, lung, skin, uterus, elastic cartilage, etc. of mammals, and is also known as elastic fiber. The elastin used in the present invention includes not only water-soluble elastin, but also hydrolyzed elastin, α-elastin, κ-elastin, elastin peptides, etc.

[0029] Elastin can be extracted from the connective tissue of animal biological tissues, for example, mammals such as pigs, cows, horses, and sheep, and fish such as bonito, tuna, yellowtail, and salmon, by hydrolyzing the elastin with acid or alkali or by enzymatic treatment. The elastin used in the present invention can be naturally derived or synthetic, and is not particularly limited, but from the viewpoint of improving vascular flexibility and vascular endothelial function, elastin derived from fish is preferred, and among fish, elastin derived from bonito or tuna is more preferred, with bonito being particularly preferred.

[0030] The proportion of elastin to be incorporated into the composition of the present invention is not particularly limited, and can be appropriately set within a wide range depending on various conditions such as the purpose, shape, and intended use. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the proportion is preferably 0.0001 to 20% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, the proportion is more preferably 0.001 to 10% by mass, and particularly preferably 0.01 to 5% by mass. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the proportion is preferably 0.00001 to 10% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, the proportion is more preferably 0.0001 to 5% by mass, and particularly preferably 0.001 to 1% by mass.

[0031] [Vitamin E] In the present invention, vitamin E can be used together with pine bark. Vitamin E used in the present invention refers to vitamin E (tocopherol, tocotrienol) and its derivatives. The type of vitamin E used in the present invention is not particularly limited. For example, tocopherol and tocotrienol may be any of α-, β-, γ-, and δ-tocopherols, and may be either d- or dl-isomers. Examples of vitamin E derivatives include organic tocopherol esters such as tocopherol acetate, tocopherol succinate, tocopherol linoleate, tocopherol linolenate, tocopherol nicotinate, and tocopherol (linoleate / oleate). From the viewpoint of improving vascular flexibility and vascular endothelial function, α-tocopherol and γ-tocopherol are preferred, with α-tocopherol being more preferred. Among α-tocopherols, the d-isomer is particularly preferred.

[0032] The proportion of vitamin E to be incorporated into the composition of the present invention is not particularly limited, and can be appropriately set within a wide range depending on various conditions such as the purpose, shape, and subject of use. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the proportion is preferably 0.0001 to 20% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, the proportion is more preferably 0.001 to 10% by mass, and particularly preferably 0.01 to 5% by mass. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the proportion is preferably 0.00001 to 10% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, the proportion is more preferably 0.0001 to 5% by mass, and particularly preferably 0.001 to 1% by mass.

[0033] The total proportion of pine bark, ginger, elastin, and vitamin E in the composition of the present invention is not particularly limited, and can be appropriately set within a wide range depending on various conditions, such as the purpose, form, and target of use. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the total proportion is preferably 0.02 to 100% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, it is more preferably 0.1 to 70% by mass, and particularly preferably 1.0 to 45% by mass. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the total proportion is preferably 0.0001 to 30% by mass, and from the viewpoints of improving vascular flexibility and vascular endothelial function, it is more preferably 0.001 to 20% by mass, and particularly preferably 0.01 to 10% by mass.

[0034] In the composition of the present invention, the ratio of ginger to pine bark is not particularly limited, but from the viewpoint of improving vascular flexibility and vascular endothelial function, the ratio of ginger to pine bark is preferably 0.0001 to 50 times, more preferably 0.0005 to 20 times, and particularly preferably 0.001 to 10 times, the mass of ginger per 1 mass of pine bark.

[0035] In the composition of the present invention, the ratio of elastin to pine bark is not particularly limited, but from the standpoint of improving vascular flexibility and vascular endothelial function, the ratio of elastin to pine bark is preferably 0.0001 to 50 times, more preferably 0.0005 to 20 times, and particularly preferably 0.001 to 10 times, the mass of which is 1 part pine bark.

[0036] In the composition of the present invention, the content ratio of vitamin E to pine bark is not particularly limited, but from the viewpoint of improving vascular flexibility and vascular endothelial function, the content ratio of vitamin E to pine bark is preferably 0.0001 to 50 times, more preferably 0.0005 to 20 times, and particularly preferably 0.001 to 10 times, the mass of pine bark per 1 mass of pine bark.

[0037] In the composition of the present invention, the ratio of the total amount of ginger, elastin, and vitamin E to the pine bark is not particularly limited, but from the standpoint of improving vascular flexibility and vascular endothelial function, the ratio of the total amount of ginger, elastin, and vitamin E to the pine bark is preferably 0.0001 to 150 times, more preferably 0.0005 to 60 times, and particularly preferably 0.001 to 30 times, the mass of pine bark being 1.

[0038] The daily intake per body weight of the composition of the present invention is not particularly limited and can be appropriately determined depending on the mode of use and the content of use by the user. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the intake is preferably 1 to 1,000 mg / kg, more preferably 3 to 200 mg / kg, based on the body weight of the user. From the viewpoint of improving vascular flexibility and vascular endothelial function, the intake is particularly preferably 5 to 100 mg / kg. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the intake is preferably 200 to 20,000 mg / kg, more preferably 600 to 15,000 mg / kg, based on the body weight of the user. From the viewpoint of improving vascular flexibility and vascular endothelial function, the intake is particularly preferably 1,000 to 10,000 mg / kg.

[0039] Similarly, the single intake amount of the composition of the present invention per body weight is not particularly limited. For example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the dose is preferably 0.3 to 1,000 mg / kg, more preferably 1.0 to 200 mg / kg, based on the user's body weight, and particularly preferably 1.6 to 100 mg / kg, from the viewpoint of improving vascular flexibility and vascular endothelial function. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the dose is preferably 66 to 20,000 mg / kg, more preferably 200 to 15,000 mg / kg, based on the user's body weight, and particularly preferably 333 to 10,000 mg / kg, from the viewpoint of improving vascular flexibility and vascular endothelial function.

[0040] The daily intake of the composition of the present invention is not particularly limited, and for example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the daily intake is preferably 0.05 to 50 g, more preferably 0.1 to 10 g, and from the viewpoint of improving vascular flexibility and vascular endothelial function, particularly preferably 0.2 to 5 g. For example, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the daily intake is preferably 10 to 1000 g, more preferably 30 to 750 g, and from the viewpoint of improving vascular flexibility and vascular endothelial function, particularly preferably 50 to 500 g.

[0041] The amount of the composition of the present invention to be taken at one time is not particularly limited, and for example, when the composition of the present invention is a solid formulation (powder, granules, tablets, capsules, chewable tablets, etc.), the amount is preferably 0.01 to 50 g, more preferably 0.03 to 10 g, and from the viewpoint of improving vascular flexibility and vascular endothelial function, particularly preferably 0.06 to 5 g. Furthermore, when the composition of the present invention is a liquid formulation (liquid, gel, paste, etc.), the amount is preferably 3.3 to 1000 g, more preferably 10 to 750 g, and from the viewpoint of improving vascular flexibility and vascular endothelial function, particularly preferably 16 to 500 g.

[0042] The composition of the present invention may contain only components selected from the group consisting of pine bark, ginger, elastin, and vitamin E, or may contain other components in addition to these. Examples of such other components include proteins, dietary fiber (e.g., soluble dietary fiber, insoluble dietary fiber), vitamins other than vitamin E, minerals, plants or plant products other than pine bark and ginger, algae, lactic acid bacteria, yeast, and other microorganisms. Furthermore, as needed, commonly used ingredients in the food industry, such as dextrin, starch, and other sugars, oligosaccharides, sweeteners, acidulants, colorants, thickeners, glazing agents, excipients, nutritional supplements, binders, lubricants, stabilizers, diluents, bulking agents, emulsifiers, food additives, and seasonings, may also be included. The content of these other components can be appropriately selected depending on the form of the composition of the present invention.

[0043] The composition of the present invention can be used, for example, as a pharmaceutical product (including quasi-drugs), a so-called health food such as a functional food whose efficacy has been approved by a designated organization, such as a food for specified health uses, a food with nutrient function claims, or a food with functional claims, or as a general food, a food additive, feed, etc. The composition of the present invention may be taken before meals, between meals, after meals, or simultaneously with meals, but from the viewpoint of the effect of improving vascular flexibility and vascular endothelial function, it is preferable to take it before meals, after meals, or simultaneously with meals, and more preferably before meals.

[0044] The composition of the present invention is not particularly limited as long as it can be distinguished from other products in terms of its use in improving vascular flexibility or vascular endothelial function. For example, the scope of the present invention includes products that display the function of improving vascular endothelial function on the body, packaging, instructions, or promotional materials of the product of the present invention. Examples of such products include so-called health foods, such as pharmaceuticals (including quasi-drugs), functional foods such as foods for specified health uses, foods with nutrient functions, and foods with functional claims that have been approved by designated organizations, and animal feeds. Examples of so-called health foods include those that display claims such as "helps maintain vascular flexibility (vasodilation after vascular constriction) that declines with age," "enhance vascular function," "maintain vascular function," "enhance vascular endothelial function," "maintain vascular endothelial function," "enhance vascular endothelial function of arteries that declines with age," "maintain vascular endothelial function of arteries that declines with age," "enhance vasodilation after vasodilation," "maintain vasodilation after vasoconstriction," "enhance vascular flexibility," "maintain vascular flexibility," "enhance vascular flexibility," "maintain vascular flexibility," and the like.

[0045] The form of the composition of the present invention is not particularly limited and can be any form. For example, forms suitable for oral use, specifically, powder, granules, tablets, liquid, gel, paste, capsules such as hard capsules and soft capsules, caplets, tablets, gels, jellies, gummies, wafers, biscuits, cookies, cakes, chewable tablets, syrups, sticks, etc., can be included. Depending on the form used, various excipients, binders, lubricants, stabilizers, diluents, bulking agents, thickeners, gelling agents, emulsifiers, colorants, flavors, sweeteners, additives, etc. can be blended. In terms of the inhibitory effect on postprandial uric acid level increase, the composition of the present invention is preferably in the form of granules, tablets, capsules, or liquid, and more preferably in the form of granules, tablets, or capsules. Here, granules refer to a composition obtained by granulating powder, and may be consumed directly or dissolved in a liquid such as water and consumed. Furthermore, the term "gel-like" refers to a composition that contains water and a gelling agent and has viscosity or elasticity.

[0046] The packaging form of the composition of the present invention is not particularly limited and can be appropriately selected depending on the dosage form, etc., and examples include blister packs such as PTPs, strip packaging, heat seals, aluminum pouches, film packaging using plastics or synthetic resins, glass containers such as vials, and plastic containers such as ampoules. [Example]

[0047] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples, and the present invention can take various forms as long as the object of the present invention can be achieved.

[0048] <Tests> Evaluation of the effects of improving vascular flexibility and vascular endothelial function In this study, the expression level of NO synthase gene was evaluated. When NO produced by NO synthase (eNOS) in the vascular endothelium diffuses and reaches vascular smooth muscle, the muscle relaxes and the blood vessels dilate. When the expression level of NO synthase gene increases, the amount of NO synthase increases, and the amount of NO produced increases, which is expected to improve vascular flexibility and vascular endothelial function. Therefore, by evaluating the expression level of NO synthase gene, it is possible to evaluate the effects of improving vascular flexibility and vascular endothelial function.

[0049] (1) Test substance The pine bark used was a hot water extract of pine bark containing OPCs (proanthocyanidins with a degree of polymerization of 2 to 4), the ginger used was crushed Kintoki ginger, the elastin used was elastin derived from bonito, and the vitamin E used was d-α-tocopherol. Chlorogenic acid was also used as a food ingredient known to improve vascular flexibility and vascular endothelial function. The pine bark, ginger, elastin, vitamin E, and chlorogenic acid were all commercially available. These ingredients were prepared in EGM-2 medium (manufactured by Lonza) in the proportions shown in Table 1 to prepare test substance-containing media for the Examples and Comparative Examples.

[0050] [Table 1]

[0051] (2)Cell culture Human umbilical vein endothelial cells (Lonza) were cultured in EGM-2 medium (Lonza). 1 × 10 cells were plated onto a collagen-coated 96-well plate. 4 The cells were seeded at 100 μL / well so that there were 10 cells / well, and the cells were pre-cultured for 24 hours in a 5% CO incubator at 37°C. After removing the medium from each well, 100 μL / well of medium containing a test substance listed in Table 1 was added, and the cells were cultured for 24 hours.

[0052] (3) Measurement of NO synthase gene expression level After removing the medium from each well and washing once with PBS, RNA was recovered using an RNeasy Mini Kit (QIAGEN). One-step real-time PCR was performed on the obtained RNA using the One Step TB Green® PrimeScript RT-PCR Kit II (Perfect Real Time). NOS3 (eNOS) gene expression levels were measured using NOS3 (eNOS) primers (Takara). GAPDH gene expression levels were measured as an endogenous control using GAPDH primers (QIAGEN).

[0053] (4) Evaluation of NO synthase gene expression levels The expression level of the NO synthase gene was calculated relative to the expression level of the group not administered the test substance (control), which was set to 1. The results are shown in FIG.

[0054] The groups administered with pine bark and at least one selected from the group consisting of ginger, elastin, and vitamin E (Examples 1 to 7) all showed an increase in NO synthase gene expression compared to the group administered with pine bark alone (Comparative Example 1). No increase in NO synthase gene expression was observed in the group administered with ginger alone (Comparative Example 2), the group administered with elastin alone (Comparative Example 3), or the group administered with vitamin E alone (Comparative Example 4). This demonstrates that the intake of a composition containing at least one selected from the group consisting of pine bark, ginger, elastin, and vitamin E increases NO synthase gene expression, thereby improving vascular flexibility and vascular endothelial function. In particular, the groups administered with two or more ingredients selected from the group consisting of pine bark, ginger, elastin, and vitamin E (Examples 4 to 7) showed a more excellent increase in NO synthase gene expression, and the group administered with pine bark, ginger, elastin, and vitamin E (Example 7) showed a significantly excellent increase in NO synthase gene expression. These results demonstrate that the composition of the present invention, containing at least one ingredient selected from the group consisting of pine bark, ginger, elastin, and vitamin E, exhibits excellent vascular flexibility-improving effects and vascular endothelial function-improving effects. Furthermore, when the composition contains two or more ingredients selected from the group consisting of pine bark, ginger, elastin, and vitamin E, the vascular flexibility-improving effects and vascular endothelial function-improving effects are more pronounced. Furthermore, when the composition contains pine bark, ginger, elastin, and vitamin E, the vascular flexibility-improving effects and vascular endothelial function-improving effects are significantly pronounced. On the other hand, although the group administered only chlorogenic acid (Comparative Example 5) showed an increase in NO synthase gene expression compared to the control, the group administered pine bark and chlorogenic acid (Comparative Example 6) did not show an increase in NO synthase gene expression compared to the group administered only pine bark (Comparative Example 1) or the group administered only chlorogenic acid (Comparative Example 5), indicating that the combination of pine bark and chlorogenic acid does not have the effect of improving vascular flexibility or vascular endothelial function.

[0055] (Manufacturing example) Based on the results of the Examples, the following Production Examples of the present invention are given below.

[0056] [Production Example 1-4: Granules] According to the formulation in Table 2, pine bark, at least one selected from the group consisting of ginger, elastin, and vitamin E, and other ingredients were mixed, and then fluidized bed granulation was performed using a granulator to produce the granules described in Production Examples 1-4. The granules described in Production Examples 1-4 only need to be taken at a dose of 3 g per day, and may be taken by dissolving in 100 ml of water or other solvent, or may be taken as is. All of the granules in Production Examples 1-4 are effective in improving vascular flexibility and vascular endothelial function.

[0057] [Table 2]

[0058] [Manufacturing Example 5-8: Tablets] As shown in Table 3, pine bark, at least one selected from the group consisting of ginger, elastin, and vitamin E, and other ingredients were mixed, and then tableted using a rotary tablet press to produce the tablets of Production Examples 5-8. The tablets were manufactured with a tablet diameter of 8 mm, a tablet thickness of 4.5 mm, a weight of 250 mg, and a hardness of 5 kgf or more. One to two tablets of Production Examples 5-8 can be taken per day with 100 ml of water, etc. All tablets of Production Examples 5-8 are effective in improving vascular flexibility and vascular endothelial function.

[0059] [Table 3]

[0060] [Manufacturing Example 9-12: Hard Capsules] As shown in Table 4, pine bark, at least one selected from the group consisting of ginger, elastin, and vitamin E, and other ingredients were mixed, and the mixture was then coated with a film containing gelatin or hydroxypropyl cellulose to produce hard capsules. Each hard capsule was 300 mg. One to two capsules per day could be taken with 100 ml of water, for example. Furthermore, all of the hard capsules in Production Examples 9-12 were effective in improving vascular flexibility and vascular endothelial function.

[0061] [Table 4]

[0062] [Manufacturing example 13-16: PET beverage] As shown in Table 5, a liquid containing pine bark, at least one selected from the group consisting of ginger, elastin, and vitamin E, and other ingredients was mixed and packed into a PET container to produce a PET beverage. Each PET beverage was produced in a 500ml bottle. One bottle should be consumed per day. Furthermore, all of the PET beverages in Production Examples 13-16 are effective in improving vascular flexibility and vascular endothelial function.

[0063] [Table 5] [Industrial Applicability]

[0064] The composition of the present invention exhibits excellent effects of improving vascular flexibility and vascular endothelial function, and is therefore highly useful industrially.

Claims

1. An oral composition for improving blood vessel flexibility, which contains pine bark as an active ingredient for improving blood vessel flexibility, and further contains elastin and vitamin E.

2. An oral composition for improving vascular endothelial function, which contains pine bark as an active ingredient for improving vascular endothelial function, and further contains elastin and vitamin E.

3. An oral composition for improving blood vessel flexibility, which contains pine bark as an active ingredient that helps maintain the flexibility of blood vessels, which decreases with age, and further contains elastin and vitamin E.

Images