JP6869712B2 - Composition for improving vascular endothelial function - Google Patents

Description

The present invention relates to a composition for improving vascular endothelial function. More specifically, the present invention relates to a composition for improving vascular endothelial function, which comprises a rice bran enzyme-treated product.

The vascular endothelium exists in the innermost layer of blood vessels, and has various vascular permeability such as blood components, contraction and relaxation reactions of blood vessels, regulation of blood coagulation by being involved in the coagulation / fibrinolytic system, control of platelet function, etc. It has a function. These functions are deeply involved in the production of various bioactive factors by vascular endothelial cells.

Nuclear factor Erythrocyte 2 related factor 2 (Nuclear factor erythroid 2-related factor 2; Nrf2) plays an important role in protecting the vascular endothelium from oxidative stress. Nrf2 normally exists in a state of being bound to the inhibitory factor Keap1, and when cells are exposed to oxidative stress or electrophiles, the inhibition by Keap1 is released, and antioxidant enzymes and the like can be induced. Are known. It is also known that Nrf2-Kep1 is stimulated by nitric oxide as well as oxidative stress, and Nrf2 is activated. Furthermore, Nrf2 is known to improve vascular endothelial function by inducing heme oxygenase (HO-1) in the vascular endothelium.

In addition, sirtuin is known to be involved in suppression of cell aging due to calorie restriction, prolongation of life span, etc., but cell aging due to oxidative stress is caused by overexpression of the sirtuin family Sirtu1 in vascular endothelial cells. It is known that induction can be avoided.

The applicant has found that pressed refined rice bran oil (Patent Document 1), extract of Seiyoyanagi (Patent Document 2), etc. obtained by steam distillation of pressed rice bran oil are useful for improving vascular endothelial function. Is found.

Japanese Unexamined Patent Publication No. 2012-206958 Japanese Unexamined Patent Publication No. 2013-173693

The present invention has been made in view of the above-mentioned current state of the prior art, and an object of the present invention is to provide a novel composition for improving vascular endothelial function capable of improving vascular endothelial function through the expression promoting action of Nrf2. And.

As a result of repeated diligent studies to achieve the above-mentioned object, the present inventors have surprisingly found that the rice bran enzyme-treated product has an excellent Nrf2 expression promoting action. Furthermore, the present inventors have also found that the rice bran enzyme-treated product has an action of promoting the expression of Sirt1. The present inventors have completed the present invention by repeating further research based on such findings.

That is, the present invention includes the subjects described in the following sections.
Item 1.
A composition for improving vascular endothelial function, which comprises a rice bran enzyme-treated product.
Item 2.
Item 2. The composition according to Item 1, wherein the rice bran enzyme-treated product is a rice bran protease-treated product.
Item 3.
Item 3. The composition according to Item 1 or 2, wherein the rice bran enzyme-treated product is a rice bran thermolysin-treated product.
Item 4.
The composition according to any one of Items 1 to 3 above, which is an oral composition.
Item 5.
The composition according to any one of Items 1 to 4, which is a food composition or a pharmaceutical composition.

According to the present invention, the vascular endothelial function can be improved.

Hereinafter, the present invention will be described in detail.

The present invention includes a composition for improving vascular endothelial function. The composition for improving vascular endothelial function of the present invention contains a rice bran enzyme-treated product. In the present specification, the composition for improving vascular endothelial function of the present invention may be simply referred to as "the composition of the present invention".

The rice bran enzyme-treated product contained in the composition of the present invention is obtained by treating rice bran with an enzyme.

The rice bran is not particularly limited, but is preferably a defatted rice bran. Further, it is preferable that the degreased rice bran is degreased by squeezing (squeezed defatted rice bran), and the lipase is inactivated by heating the rice bran (raw rice bran) obtained after milling, and the squeezing obtained by further squeezing treatment. Solvent degreased rice bran is more preferable. Further, defatted rice bran having a lipid content of 5 to 20% by mass is preferable. A defatted rice bran having a lipid content in this range is preferably obtained by squeezing defatting (particularly, squeezing defatting after lipase deactivation treatment).

The lipase inactivation treatment is carried out for the purpose of preventing oxidative deterioration of rice bran (raw rice bran) containing about 18 to 20% by mass of lipid. Usually, it is carried out by heating and roasting raw rice bran at about 70 to 130 ° C. The squeezing treatment is carried out by a known squeezing method, for example, rice bran that has been heat roasted and has reached about 100 to 115 ° C. is squeezed by a low temperature continuous squeezing machine (for example, a miracle chamber sold by TechnoSigma). The squeezing is preferably carried out until the lipid content in the defatted rice bran after squeezing is about 5 to 20% by weight. Further, the drying treatment may be carried out for the purpose of reducing the water content of the rice bran before the lipase deactivation treatment and the degreased rice bran after the oil extraction. Further, it is preferable that the defatted rice bran obtained by the pressing treatment is further crushed and classified by a known method to be powdered. At this time, the average particle size of the powder is preferably 5 to 200 μm, more preferably 10 to 150 μm, and most preferably 40 to 100 μm. The lipid content in the defatted rice bran is preferably 5 to 20% by weight, more preferably 7 to 15% by weight, as described above. It is also possible to purchase and use a commercially available degreased rice bran as described above. Examples of such commercially available degreased rice bran include Nucup (manufactured by Tokyo Blef Co., Ltd.) and Hi-Bref (manufactured by Saint-Blanc Co., Ltd.).

The rice bran in the present application also includes a water extract of rice bran. The water extract of rice bran is obtained by extracting the rice bran with water. The water extract of rice bran is preferably a liquid composition (composed of water and the extracted rice bran component) obtained by extracting the rice bran with water. A rice bran enzyme-treated product can also be obtained by treating the water extract of rice bran with an enzyme described later. The water extraction conditions are not particularly limited, and examples thereof include a method of immersing (or stirring) rice bran in water at about 10 to 50 ° C. for about 1 to 24 hours.

The enzyme used for preparing the rice bran enzyme-treated product is not particularly limited as long as it is a proteolytic enzyme (protease: including peptidase) capable of degrading proteins and polypeptides contained in rice bran, and a known enzyme is used. be able to. Examples of such enzymes include trypsin (EC 3.4.21.4), papain (EC 3.4.22.2), thermolysin (EC 3.4.24.27), and chymotrypsin (EC 3.4.21.). 1, EC 3.4.21.2.) And the like. The description in parentheses is the enzyme number (Enzyme Commission numbers). Of these, thermolysin is preferred. In this specification, the rice bran enzyme-treated product may be referred to as, for example, a rice bran protease treated product, a rice bran thermolysin-treated product, a rice bran chymotrypsin-treated product, or the like, depending on the enzyme used for the enzyme treatment.

As the enzyme used for the enzyme treatment of rice bran, for example, a commercially available product can be purchased and used. For example, thermolysin can be purchased from Peptide Institute, Ltd., Yamato Kasei Co., Ltd., Nacalai Tesque Co., Ltd., Amano Enzyme, Wako Pure Chemical Industries, Ltd., etc.

The enzyme treatment conditions for obtaining the rice bran enzyme-treated product are not particularly limited as long as the activity of the enzyme to be used can be obtained, and can be appropriately determined. For example, when thermolysin is used as an enzyme, the treatment can be carried out by mixing rice bran and thermolysin with water and reacting them at 30 ° C. to 70 ° C. with gentle stirring for about 1 to 30 hours. The optimum conditions for the reaction temperature and reaction time differ depending on the structure and function of the manufacturing equipment used and the purity of the enzyme used. For example, the optimum temperature of thermolysin is around 65 ° C., but industrially, it is preferable to determine the reaction temperature around 35 to 60 ° C. (more preferably 50 to 60 ° C.). After the treatment, the enzyme may be inactivated for the purpose of stabilizing the quality. The enzyme inactivation treatment is performed by heating, for example (conditions differ depending on the enzyme used, but for example, when an enzyme preparation containing about 3% thermolysin is used, the treatment is performed at 70 to 100 ° C. for about 1 to 20 minutes. Can be adopted) and so on. Further, after the treatment, the filtrate or the like may be recovered by filtration, centrifugation or the like, or the obtained filtrate or the like may be purified by concentration, drying, column treatment or the like.

As described above, the rice bran enzyme-treated product is obtained by enzymatically treating rice bran, and the rice bran enzyme-treated product contains enzymatic decomposition products of proteins and polypeptides contained in rice bran.

The composition of the present invention may be the above-mentioned rice bran processed product itself, or may contain other components in addition to the rice bran enzyme treated product. Other ingredients include pharmaceutically or food hygiene-acceptable carriers and the like.

The composition of the present invention can be used as an oral composition. As the oral composition, for example, it can be used as a food composition, a pharmaceutical composition (including quasi-drugs), and the like. Further, the rice bran enzyme-treated product contained in the composition of the present invention not only has an excellent Nrf2 expression-promoting action, but also has a Sirtuin expression-promoting action, and therefore is preferably used as a composition for improving vascular endothelial function. Can be done. Furthermore, since the composition of the present invention can improve vascular endothelial function, it can be used for diseases associated with a decrease in vascular endothelial function (for example, hypertension, arteriosclerosis, dyslipidemia, diabetes, diabetic complications, etc.). It can be preferably used for prevention and / or treatment. Further, for the purpose of alleviating the onset and symptoms of these diseases, it can be preferably used in applications for people who develop these diseases or who have a high risk of developing them.

The composition of the present invention can be ingested once or in a plurality of times (preferably 2 to 3 times). In addition, although humans are preferable as the ingestion target, non-human mammals other than humans may be used.

Further, since the rice bran enzyme-treated product contained in the composition of the present invention has an excellent Nrf2 expression promoting action, it can exert an antioxidant action, more specifically, an antioxidant action on the vascular endothelium. The composition of the present invention can be preferably used for the purpose of such an action. For example, the composition of the present invention can be used for antioxidants, more specifically for antioxidants of vascular endothelium.

Furthermore, since the rice bran enzyme-treated product contained in the composition of the present invention has an action of promoting the expression of Sirtu1, it is possible to exert actions such as anti-cell aging action and anti-arteriosclerosis action, and the composition of the present invention can exert an action. It can be preferably used for applications aimed at such an action. For example, the composition of the present invention can be used for anti-aging, anti-arteriosclerosis and the like.

The form of the composition of the present invention is not particularly limited, but when it is used as an oral composition, it may be in the form of, for example, a hard capsule, a soft capsule, a supplement, a chewable tablet, a beverage, a powdered beverage, granules, or a film. it can. When used as a food or drink, for example, a concentrated type that is diluted with tea-based beverages, sports beverages, beauty beverages, fruit juice beverages, carbonated beverages, alcoholic beverages, soft drinks, jelly beverages, water, hot water, carbonated water, etc. Beverages such as beverages, powders and granules to be drunk by dissolving or suspending in water or hot water, dried solids such as tablets, tablet confectionery, jellies, snacks, baked confectionery, fried confectionery, cakes, chocolate, It can also be in the form of foods such as sweets such as gum, candy and gummy, soups, noodles, rice and cereals. Of these, supplement types, chewable tablets, one-shot drink types, etc. are preferable when used in normal life, and beverage forms such as sports drinks are most preferable when ingested for the purpose of enhancing exercise effects. .. In addition, these oral compositions can be provided to consumers as packaged foods filled in containers. The container is not particularly limited as long as it can be sealed.

In the case of oral solid preparations such as tablets, granules and capsules, and oral liquid preparations such as oral liquids and syrups, the composition of the present invention is 0. It may be blended in an amount of about 01 to 95% by mass, preferably about 5 to 90% by mass, and most preferably about 10 to 80% by mass. In the case of an oral composition other than the above, the composition of the present invention may be blended in an amount of about 0.001 to 50% by mass, preferably about 0.005 to 30% by mass, and most preferably about 0.01 to 10% by mass. Good.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the following examples.

Production example: Preparation of rice bran enzyme-treated product In this production example, an enzyme preparation containing Hybref (manufactured by Saint-Blanc) as rice bran and 3% thermolysin as an enzyme was used. The high bref is a squeezed degreased rice bran that has been degreased by squeezing the rice bran. Rice bran was mixed with water to a concentration of 30 g / L, and thermolysin was added to the mixture to a concentration of 0.0045 g / L. Then, the mixture was stirred at 55 ° C. for 15 hours. Then, the mixture was stirred at 80 ° C. for 5 minutes to inactivate the enzyme. Then, the obtained treatment liquid was filtered using diatomaceous earth and then concentrated, and the concentrated liquid was further sterilized at 121 ° C. for 15 minutes. The sterilized filtrate thus obtained was freeze-dried to obtain a rice bran enzyme-treated product.

Test Example: The rice bran enzyme-treated product prepared in the above production example was used as a sample for examining the effects of Nrf2 and Sirtu1 on gene expression. H ₂ O ₂ and resveratrol, as well as DMSO were used as controls. Human umbilical vein endothelial cells (HUVECs, manufactured by Sanko Junyaku Co., Ltd.) were cultured in 800 μL of 2% FBS medium using a 12-well plate, and various samples were adjusted to 5 times the final concentration shown in Table 1 below. 200 μL of dissolved 2% FBS medium was added and cultured for 6 hours and 24 hours. The Total RNA was then extracted using the Total RNA Mini Kit. Single-strand cDNA (ssDNA) was synthesized from 0.35 μg of Total RNA using the PrimeScript RT reagent Kit. When the amount of total RNA recovered was insufficient, the dilution rate of ssDNA was changed according to the amount of recovery, and then the mRNA was subjected to quantitative analysis. Quantitative analysis of Nrf2 and Sirtuin mRNAs was performed using a real-time PCR system (Applied Biosystems). In addition, Premix Ex Taq (manufactured by Takara Bio Inc.), Assay-on-Demand, and Gene Expression Products (Hs00232352_m1 for NFE2L2 (Nrf2), Hs0109005_m1 for Sirt1) Quantitatively, Hs0109005_m1 for Quantitative data were corrected by ΔΔCt method at the expression level of Ribosomal protein S18 (RPS18), respectively. The results are shown in Table 1 below.

From Table 1, it was found that the rice bran enzyme-treated product had an excellent Nrf2 expression promoting action and a Sirt1 expression promoting action. From these results, it is considered that the rice bran enzyme-treated product exerts an excellent vascular endothelial function improving action, an antioxidant action, an anti-arteriosclerosis action, and the like by promoting the expression of Nrf2 and Sirtu1.

Claims (3)

A composition for improving vascular endothelial function, which comprises a thermolysin-treated product of rice bran. It is an oral composition, The composition of claim 1. The composition according to claim 1 or 2 , which is a food composition or a pharmaceutical composition.