JP4083111B2 - β-endorphin production promoter - Google Patents
β-endorphin production promoter Download PDFInfo
- Publication number
- JP4083111B2 JP4083111B2 JP2003424336A JP2003424336A JP4083111B2 JP 4083111 B2 JP4083111 B2 JP 4083111B2 JP 2003424336 A JP2003424336 A JP 2003424336A JP 2003424336 A JP2003424336 A JP 2003424336A JP 4083111 B2 JP4083111 B2 JP 4083111B2
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- JP
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- Prior art keywords
- endorphin
- production
- extract
- production promoter
- terminaria
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
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Description
本発明は、β−エンドルフィンの産生を促進するβ−エンドルフィン産生促進剤に関する。さらに詳しくは、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を含有するβ−エンドルフィン産生促進剤に関する。 The present invention relates to a β-endorphin production promoter that promotes the production of β-endorphin. More specifically, the present invention relates to a β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp.
β−エンドルフィンは、脳下垂体の副腎皮質刺激ホルモン（ＡＣＴＨ）及びメラニン細胞刺激ホルモン（ＭＳＨ）と共通の前駆体タンパク質であるプレプロオピオメラノコルチン（ＰＯＭＣ）から生合成される内因性モルヒネ様ペプチド（オピオタイドペプチド）の一種であり、鎮痛作用や抗ストレス作用を有することから脳内快楽物質として知られている。β−エンドルフィンは、脳下垂体中葉・後葉に多く含まれ、ストレスなどの侵害要因によって血中にも分泌されることが知られているが、近年の研究によって、皮膚においてもＰＯＭＣが合成され、表皮ケラチノサイトよりβ−エンドルフィンが遊離することが明らかとされている（非特許文献１，２参照）。 β-endorphin is an endogenous morphine-like peptide (opio) that is biosynthesized from preproopiomelanocortin (POMC), a precursor protein in common with pituitary adrenocorticotropic hormone (ACTH) and melanocyte stimulating hormone (MSH). It is a kind of tide peptide) and is known as a pleasure substance in the brain because it has analgesic action and anti-stress action. β-endorphin is abundantly contained in the pituitary midlobe and posterior lobe, and is known to be secreted into the blood by noxious factors such as stress, but recent research has synthesized POMC in the skin, It has been clarified that β-endorphin is released from epidermal keratinocytes (see Non-Patent Documents 1 and 2).
しかし、β−エンドルフィンの皮膚における生理的作用については、未だ不明な点も多く、詳細な検討はなされていない現状であった。そこで、本発明者らは、ストレスによって分泌されるβ−エンドルフィンが皮膚に対して何らかの有利な作用を発揮しているのではないかとの仮説のもとに種々の検討を行った。 However, the physiological action of β-endorphin on the skin is still unclear and has not been studied in detail. Therefore, the present inventors have made various studies under the hypothesis that β-endorphin secreted by stress may exert some advantageous action on the skin.
これらの検討の結果、本発明者らは、β−エンドルフィンが皮膚において細胞賦活作用と美白作用を発揮することを見出した。 As a result of these studies, the present inventors have found that β-endorphin exhibits cell activation and whitening effects in the skin.
β−エンドルフィンが皮膚において細胞賦活作用と美白作用を発揮することが明らかとなったことから、β−エンドルフィンの産生を促進することにより、細胞賦活作用や美白作用による皮膚症状の予防や改善が可能であることが示された。このため、本発明者らはβ−エンドルフィンの産生を促進する成分を見出し、種々の皮膚症状に応用することが可能なβ−エンドルフィン産生促進剤を提供することを目的に種々の検討を行った。 Since β-endorphin has been shown to exert cell activation and whitening effects in the skin, it is possible to prevent and improve skin symptoms due to cell activation and whitening by promoting the production of β-endorphin. It was shown that. For this reason, the present inventors have found a component that promotes the production of β-endorphin, and have made various studies for the purpose of providing a β-endorphin production promoter that can be applied to various skin conditions. .
したがって、本発明の目的は、β−エンドルフィンの産生を促進する成分を見出し、種々の皮膚症状に応用することが可能なβ−エンドルフィン産生促進剤を提供することにある。 Accordingly, an object of the present invention is to provide a β-endorphin production promoter capable of finding a component that promotes the production of β-endorphin and applying it to various skin conditions.
優れた効果を発揮するβ−エンドルフィン産生促進剤を見出すために、本発明者らは、天然由来の種々の成分に関して、β−エンドルフィンの産生促進作用についての検討を行った。その結果、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより得られる抽出物に優れたβ−エンドルフィン産生促進作用を見出し、さらに種々の検討を行い、本発明を完成するに至った。 In order to find a β-endorphin production promoter exhibiting an excellent effect, the present inventors examined the production promotion action of β-endorphin with respect to various naturally derived components. As a result, an excellent β-endorphin production-promoting action was found in the extract obtained from Terminaria, Hypericum perforatum, Ginseng, Giant kelp, and various studies were conducted to complete the present invention.
すなわち、本発明は、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を含有するβ−エンドルフィン産生促進剤に関する。 That is, this invention relates to the beta-endorphin production promoter containing 1 type, or 2 or more types of extracts chosen from Terminaria, Hypericum perforatum, Periwinkle, Giant kelp.
本発明によれば、優れた効果を発揮するβ−エンドルフィン産生促進剤を得ることができる。得られたβ−エンドルフィン産生促進剤は、皮膚におけるβ−エンドルフィンの産生を促進することにより、皮膚にリラックス効果を与えるとともに、ストレスに起因する種々の皮膚症状を予防あるいは改善することが可能である。特に、細胞賦活作用や美白作用に基づく皮膚症状の予防や改善を図ることができる。本発明により予防や改善が図られる皮膚症状の例としては、しわ，たるみ，しみ，くすみ，乾燥，肌荒れなどを挙げることができる。 According to the present invention, a β-endorphin production promoter that exhibits excellent effects can be obtained. The obtained β-endorphin production promoter promotes the production of β-endorphin in the skin, thereby giving a relaxing effect to the skin and preventing or improving various skin symptoms caused by stress. . In particular, it is possible to prevent or improve skin symptoms based on cell activation and whitening. Examples of skin symptoms that can be prevented or ameliorated by the present invention include wrinkles, sagging, spots, dullness, dryness, and rough skin.
本発明の原料として用いられる植物としては、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプを挙げることができる。テルミナリア（Ｔｅｒｍｉｎａｌｉａ ｓｅｒｉｃｅａ）は、シクンシ科モモタマナ属の植物であり、セイヨウオトギリソウ（Ｈｙｐｅｒｉｃｕｍ ｐｅｒｆｏｒａｔｕｍ）は、オトギリソウ科オトギリソウ属の植物である。トウキンセンカ（Ｃａｌｅｎｄｕｌａ ｏｆｆｉｃｉｎａｌｉｓ）は、キク科カレンデュラ属の植物であり、ジャイアントケルプとは、レッソニア科に属するマクロシスティス・ピリフェラ(Ｍａｃｒｏｃｙｓｔｉｓ ｐｙｒｉｆｅｒａ)、マクロシスティス・インテグリフォリア（Ｍａｃｒｏｃｙｓｔｉｓ ｉｎｔｅｇｒｉｆｏｌｉａ）、ネオシティス・ルエトケアーナ(Ｎｅｒｅｏｃｙｓｔｉｓ ｌｕｅｔｋｅａｎａ)などの褐藻である。 Examples of the plant used as a raw material of the present invention include Terminaria, Hypericum perforatum, Ginseng, Giant kelp. Terminaria ( Terminalia sericea ) is a plant belonging to the genus Motamamana , and Hypericum perforatum is a plant belonging to the genus Hypericum . Deng Calendula (Calendula officinalis) is a Asteraceae Calendula plant of the genus, the giant kelp, macro cis-Sevilla-Pirifera belonging to Lessonia family (Macrocystis pyrifera), macro-cis Sevilla integrase Li Folia (Macrocystis integrifolia), Neoshitisu - It is a brown algae such as Ruetkeana ( Neerocystis luetkeana ).
これらの植物を使用する際は、抽出物を用いるのが簡便である。抽出には、植物の全草，花，茎，樹皮，根，芽，種子あるいは胞子等のいずれの部位を用いても構わないが、有効性や簡便性の点から、テルミナリアの場合は、樹皮あるいは根を用いるのが望ましく、セイヨウオトギリソウの場合は、全草を用いるのが望ましく、トウキンセンカの場合は、頭花を用いるのが望ましく、ジャイアントケルプの場合は、全藻を用いるのが望ましい。抽出の際は、生のまま用いてもよいが、抽出効率を考えると、細切，乾燥，粉砕等の処理を行った後に抽出を行うことが好ましい。抽出は、抽出溶媒に浸漬するか、超臨界流体や亜臨界流体を用いた抽出方法でも行うことができる。抽出効率を上げるため、撹拌や抽出溶媒中でホモジナイズしてもよい。抽出温度としては、５℃程度から抽出溶媒の沸点以下の温度とするのが適切である。抽出時間は抽出溶媒の種類や抽出温度によっても異なるが、１時間〜１４日間程度とするのが適切である。 When using these plants, it is convenient to use an extract. For extraction, any part of the plant such as whole grass, flowers, stems, bark, roots, buds, seeds or spores may be used, but from the viewpoint of effectiveness and convenience, in the case of Terminaria, the bark is used. Alternatively, it is desirable to use roots. In the case of Hypericum perforatum, it is desirable to use whole grass. In the extraction, it may be used as it is, but considering the extraction efficiency, it is preferable to perform the extraction after performing processing such as shredding, drying, and pulverization. The extraction can be performed by immersing in an extraction solvent or by an extraction method using a supercritical fluid or a subcritical fluid. In order to increase the extraction efficiency, homogenization may be performed in stirring or an extraction solvent. The extraction temperature is suitably about 5 ° C. to the boiling point of the extraction solvent. The extraction time varies depending on the type of extraction solvent and the extraction temperature, but it is appropriate to set it to about 1 hour to 14 days.
抽出溶媒としては、水の他、メタノール，エタノール，プロパノール，イソプロパノール等の低級アルコール、１，３−ブチレングリコール，プロピレングリコール，ジプロピレングリコール，グリセリン等の多価アルコール、エチルエーテル，プロピルエーテル等のエーテル類、酢酸ブチル，酢酸エチル等のエステル類、アセトン，エチルメチルケトン等のケトン類などの溶媒を用いることができ、これらより１種又は２種以上を選択して用いる。また、生理食塩水，リン酸緩衝液，リン酸緩衝生理食塩水等を用いてもよい。さらに、水や二酸化炭素，エチレン，プロピレン，エタノール，メタノール，アンモニアなどの１種又は２種以上の超臨界流体や亜臨界流体を用いてもよい。 Extraction solvents include water, lower alcohols such as methanol, ethanol, propanol, and isopropanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, dipropylene glycol, and glycerin, and ethers such as ethyl ether and propyl ether. , Solvents such as esters such as butyl acetate and ethyl acetate, and ketones such as acetone and ethyl methyl ketone can be used, and one or more of these are selected and used. Further, physiological saline, phosphate buffer, phosphate buffered saline, or the like may be used. Furthermore, you may use 1 type, or 2 or more types of supercritical fluids and subcritical fluids, such as water, a carbon dioxide, ethylene, propylene, ethanol, methanol, ammonia.
各植物の上記溶媒による抽出物は、そのままでも使用することができるが、濃縮，乾固した物を水や極性溶媒に再度溶解したり、或いはこれらの生理作用を損なわない範囲で脱色，脱臭，脱塩等の精製処理を行ったり、カラムクロマトグラフィー等による分画処理を行った後に用いてもよく、前記抽出物やその処理物及び分画物は、各処理及び分画後に凍結乾燥し、用時に溶媒に溶解して用いることもできる。 Extracts of the above-mentioned solvents of each plant can be used as they are, but the concentrated and dried products are dissolved again in water or a polar solvent, or decolorized, deodorized as long as these physiological functions are not impaired, It may be used after performing purification treatment such as desalting or fractionation by column chromatography, etc., and the extract and its treated product and fraction are lyophilized after each treatment and fractionation, It can also be used by dissolving in a solvent at the time of use.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物は、優れたβ−エンドルフィン産生促進作用を有し、β−エンドルフィン産生促進剤として利用することができる。 One or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp have an excellent β-endorphin production promoting action and can be used as a β-endorphin production promoter.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤は、単独でも使用することが出来るが、β−エンドルフィン産生促進剤として医薬品，医薬部外品，飲食品，化粧品などの種々の組成物に配合することにより、β−エンドルフィン産生促進作用を有する組成物を得ることが出来る。 A β-endorphin production promoter comprising one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp as an active ingredient can be used alone, but a β-endorphin production promoter As described above, a composition having a β-endorphin production promoting action can be obtained by blending with various compositions such as pharmaceuticals, quasi drugs, foods and drinks, and cosmetics.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤を配合する組成物の剤型は任意であるが、組成物が皮膚外用剤，洗浄剤，浴用剤などの場合には、ローションなどの可溶化系，乳液などの乳化系，カラミンローションなどの分散系，噴射剤と共に充填したエアゾール，軟膏剤，粉末，顆粒などの種々の剤型として提供することができる。また、組成物が経口用医薬品や飲食品の場合には、ドリンク剤・点滴剤などの液剤，ガム・飴のような固形剤，カプセル，粉末，顆粒，錠剤などの一般的な剤型とすることができる。 The form of the composition containing the β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant Kelp as an active ingredient is arbitrary. In the case of external preparations for skin, cleaning agents, bath preparations, etc., solubilizing systems such as lotions, emulsifying systems such as emulsions, dispersing systems such as calamine lotions, aerosols filled with propellants, ointments, powders, granules It can be provided as various dosage forms. In addition, when the composition is an oral drug or food and drink, it should be a general dosage form such as liquid preparations such as drinks and drops, solid preparations such as gums and candy, capsules, powders, granules and tablets. be able to.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤を配合する組成物には、これらの他に、必要に応じて、通常医薬品，医薬部外品，皮膚化粧料，浴用剤，洗浄剤，食品などに配合される油性成分，保湿剤，粉体，色素，乳化剤，可溶化剤，洗浄剤，紫外線吸収剤，増粘剤，薬剤，香料，樹脂，アルコール類，栄養強化物質，調味料などを適宜配合することができ、さらに他のβ−エンドルフィン産生促進剤との併用も可能である。 In addition to these, a composition containing a β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp as an active ingredient Ordinary pharmaceuticals, quasi-drugs, skin cosmetics, bath preparations, cleaning agents, oily ingredients blended in foods, moisturizers, powders, pigments, emulsifiers, solubilizers, cleaning agents, UV absorbers, A sticking agent, a drug, a fragrance, a resin, an alcohol, a nutrition enhancing substance, a seasoning, and the like can be blended as appropriate, and can also be used in combination with other β-endorphin production promoters.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤の種々の組成物への配合量は、組成物の種類や目的等によって調整することができるが、有効性や使用性などの点から、組成物の全量に対して０．０００１〜７５重量％が好ましく、より好ましくは０．００１〜５０重量％であり、最も好ましくは０．０１〜２５重量である。 The amount of β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp as an active ingredient is the type and purpose of the composition. However, it is preferably 0.0001 to 75% by weight, more preferably 0.001 to 50% by weight with respect to the total amount of the composition in terms of effectiveness and usability. Preferably it is 0.01-25 weight.
テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤、あるいは該β−エンドルフィン産生促進剤を配合したβ−エンドルフィン産生促進用組成物は、ストレスに起因する種々の皮膚症状を予防あるいは改善することが可能である。特に、細胞賦活作用や美白作用に基づく皮膚症状の予防や改善を図ることができ、その例としては、しわ，たるみ，しみ，くすみ，乾燥，肌荒れなどの皮膚症状を挙げることができる。 Β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp as an active ingredient, or β-endorphin production promotion containing the β-endorphin production promoter The composition for use can prevent or ameliorate various skin symptoms caused by stress. In particular, it is possible to prevent or improve skin symptoms based on cell activation and whitening effects, and examples include skin symptoms such as wrinkles, sagging, spots, dullness, dryness, and rough skin.
以下に、抽出物の製造例、作用を評価するための試験について詳細に説明するが、本発明の技術的範囲はこれによってなんら限定されるものではない。 Hereinafter, the production example of the extract and the test for evaluating the action will be described in detail, but the technical scope of the present invention is not limited thereto.
［製造例１］ テルミナリア抽出物の調製
テルミナリアの樹皮及び根１ｋｇにメタノールを９リットル加え、室温で７日間浸漬した。抽出液をろ過して回収し、溶媒を除去した後、抽出乾燥物を得た。得られた抽出乾燥物を０．８重量％となるように１，３−ブチレングリコール水溶液に溶解し、テルミナリア抽出物を得た。
[Production Example 1] Preparation of terminaria extract 9 liters of methanol was added to 1 kg of terminaria bark and roots, and immersed for 7 days at room temperature. The extract was collected by filtration, and after removing the solvent, a dried extract was obtained. The obtained dried extract was dissolved in 1,3-butylene glycol aqueous solution so as to be 0.8% by weight to obtain a terminaria extract.
［製造例２］ セイヨウオトギリソウ抽出物の調製
セイヨウオトギリソウの全草１ｋｇに５０重量％１，３−ブチレングリコール水溶液を９リットル加え、室温で７日間浸漬した。抽出液をろ過して回収し、溶媒を除去した後、抽出乾燥物を得た。得られた抽出乾燥物を２．０重量％となるように５０重量％１，３−ブチレングリコール水溶液に溶解し、セイヨウオトギリソウ抽出物を得た。
[Production Example 2] Preparation of Hypericum perforatum extract 9 kg of a 50 wt% aqueous 1,3-butylene glycol solution was added to 1 kg of whole Hypericum perforatum and soaked at room temperature for 7 days. The extract was collected by filtration, and after removing the solvent, a dried extract was obtained. The obtained dried extract was dissolved in 50 wt% 1,3-butylene glycol aqueous solution so as to be 2.0 wt% to obtain a hypericum perforatum extract.
［製造例３］ トウキンセンカ抽出物の調製
トウキンセンカの花１ｋｇに５０重量％エタノール水溶液を９リットル加え、室温で７日間浸漬した。抽出液をろ過して回収し、溶媒を除去した後、抽出乾燥物を得た。得られた抽出乾燥物を２．０重量％となるように５０重量％１，３−ブチレングリコール水溶液に溶解し、トウキンセンカ抽出物を得た。
[Manufacturing Example 3] Preparation of a milkflower extract 9 kg of 50 wt% ethanol aqueous solution was added to 1 kg of a flower of milkflower, and immersed for 7 days at room temperature. The extract was collected by filtration, and after removing the solvent, a dried extract was obtained. The obtained dried extract was dissolved in 50% by weight 1,3-butylene glycol aqueous solution so as to be 2.0% by weight, to obtain a milk extract.
［製造例４］ ジャイアントケルプ抽出物の調製
ジャイアントケルプ（Ｍａｃｒｏｃｙｓｔｉｓ ｐｙｒｉｆｅｒａ）の全草１ｋｇに３％塩化ナトリウム水溶液を９リットル加え、室温でホモジナイズした。抽出液をろ過して回収し、溶媒を除去した後、抽出乾燥物を得た。
[Production Example 4] Preparation of giant kelp extract 9 liters of a 3% aqueous sodium chloride solution was added to 1 kg of whole plant of giant kelp ( Macrocystis pyrifera ) and homogenized at room temperature. The extract was collected by filtration, and after removing the solvent, a dried extract was obtained.
［β−エンドルフィン産生促進作用の評価］
試料には、製造例１〜４にて調製したテルミナリア抽出物（試料１）、セイヨウオトギリソウ抽出物（試料２）、トウキンセンカ抽出物（試料３）、及びジャイアントケルプ抽出物（試料４）を用いた。評価は、以下の手順で行った。ヒト表皮細胞を１ウェル当たり２．０×１０４個となるように９６穴マイクロプレートに播種した。播種培地には、市販のダルベッコ改変イーグル培地（ＤＭＥＭ）に５％のウシ胎児血清を添加したものを用いた。２４時間培養後、任意の濃度の試料を添加した試験培地に交換し、さらに４８時間培養した。培養後に培養上清中に分泌されたβ−エンドルフィン産生量をＥＬＩＳＡ法により定量した。また、同時に細胞数を計測し、細胞当たりのβ−エンドルフィン産生量を算出した。各試料の評価結果を、ブランクの細胞あたりのβ−エンドルフィン産生量を１００とした場合の相対値にて表１及び表２に示す。なお、表中の**は、ｔ検定における有意確率Ｐ値に対し、有意確率１％未満の危険率（Ｐ＜０．０１）で有意差が認められたものを表したものである。
[Evaluation of β-endorphin production promoting effect]
For the samples, the terminaria extract (sample 1), Hypericum perforatum extract (sample 2), pearl millet extract (sample 3), and giant kelp extract (sample 4) prepared in Production Examples 1 to 4 were used. It was. The evaluation was performed according to the following procedure. Human epidermal cells were seeded in a 96-well microplate so as to be 2.0 × 10 4 cells per well. As the seeding medium, a commercially available Dulbecco's modified Eagle medium (DMEM) with 5% fetal bovine serum added was used. After culturing for 24 hours, the culture medium was replaced with a test medium to which a sample having an arbitrary concentration was added, and further cultured for 48 hours. The amount of β-endorphin produced secreted into the culture supernatant after culturing was quantified by ELISA. At the same time, the number of cells was counted, and the amount of β-endorphin produced per cell was calculated. The evaluation results of each sample are shown in Tables 1 and 2 as relative values when the β-endorphin production amount per blank cell is 100. In addition, ** in the table indicates that a significant difference was recognized with a risk rate (P <0.01) having a significance probability of less than 1% with respect to the significance probability P value in the t test.
表１及び表２より、試料を添加した培地において、有意なβ−エンドルフィン産生促進作用が認められた。特に、試料１〜３を０．２５〜１．０％添加した場合、及び試料４を０．２５〜１．０ｍｇ／ｍＬ添加した場合には、ブランクと比較して、危険率１％未満で有意なβ−エンドルフィン産生促進作用が認められた。このことから、β−エンドルフィンは、優れたβ−エンドルフィン産生促進作用を有することが明らかとなった。 From Tables 1 and 2, a significant β-endorphin production promoting action was observed in the medium to which the sample was added. In particular, when samples 1 to 3 were added at 0.25 to 1.0% and sample 4 was added at 0.25 to 1.0 mg / mL, the risk rate was less than 1% compared to the blank. A significant β-endorphin production promoting action was observed. From this, it was revealed that β-endorphin has an excellent β-endorphin production promoting action.
次に、本発明者らが明らかにしたβ−エンドルフィンの皮膚における生理的作用の検討について示す。 Next, examination of the physiological action of β-endorphin on the skin, which has been clarified by the present inventors, will be described.
［β−エンドルフィンによる表皮細胞賦活作用の評価］
評価は、試料としてヒトβ−エンドルフィンを用い、以下の手順で行った。ヒト表皮細胞を１ウェル当たり２．０×１０４個となるように９６穴マイクロプレートに播種した。播種培地には、市販のダルベッコ改変イーグル培地（ＤＭＥＭ）に５％のウシ胎児血清を添加したものを用いた。２４時間培養後、任意の濃度の試料を添加した試験培地に交換し、さらに２４時間培養した。次いで３−(４,５−ジメチル−２−チアゾリル)−２,５−ジフェニルテトラゾリウムブロミド（ＭＴＴ）を１００μｇ／ｍＬ含有する培地に交換して２時間培養し、テトラゾリウム環の開環により生じるフォルマザンを２−プロパノールにて抽出し、マイクロプレートリーダーにて５５０ｎｍの吸光度を測定した。同時に濁度として６５０ｎｍにおける吸光度を測定し、両測定値の差により細胞賦活作用を評価した。評価結果を、試料無添加のブランクにおける細胞賦活作用を１００とした場合の相対値にて表３に示す。なお、表中の**は、ｔ検定における有意確率Ｐ値に対し、有意確率１％未満の危険率（Ｐ＜０．０１）で有意差が認められたものを表したものである。
[Evaluation of epidermal cell activation by β-endorphin]
Evaluation was performed in the following procedure using human β-endorphin as a sample. Human epidermal cells were seeded in a 96-well microplate so as to be 2.0 × 10 4 cells per well. As the seeding medium, a commercially available Dulbecco's modified Eagle medium (DMEM) with 5% fetal bovine serum added was used. After culturing for 24 hours, the culture medium was replaced with a test medium to which a sample having an arbitrary concentration was added, and further cultured for 24 hours. Next, the medium containing 100 μg / mL of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) was exchanged and cultured for 2 hours. Formazan generated by the opening of the tetrazolium ring was removed. Extraction was performed with 2-propanol, and absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. The evaluation results are shown in Table 3 as relative values when the cell activation effect in the blank with no sample added is taken as 100. In addition, ** in the table indicates that a significant difference was recognized with a risk rate (P <0.01) having a significance probability of less than 1% with respect to the significance probability P value in the t-test.
表３より明らかなように、β−エンドルフィンを添加した培地において、有意な表皮細胞賦活作用が認められた。特に、β−エンドルフィンを０．１２５〜０．５ｎｇ／ｍＬ添加した場合には、ブランクと比較して、危険率１％未満で有意な表皮細胞賦活作用が認められた。このことから、β−エンドルフィンは、優れた表皮細胞賦活作用を有することが明らかとなった。 As is clear from Table 3, a significant epidermal cell activation effect was observed in the medium supplemented with β-endorphin. In particular, when β-endorphin was added in an amount of 0.125 to 0.5 ng / mL, a significant epidermal cell activation effect was observed at a risk rate of less than 1% compared to the blank. From this, it was revealed that β-endorphin has an excellent epidermal cell activation effect.
［β−エンドルフィンによる真皮線維芽細胞賦活作用の評価］
評価は、試料としてヒトβ−エンドルフィンを用い、以下の手順で行った。正常ヒト真皮線維芽細胞を１ウェル当たり２．０×１０４個となるように９６穴マイクロプレートに播種した。播種培地には、ダルベッコ改変イーグル培地（ＤＭＥＭ）に１％のウシ胎児血清を添加したものを用いた。２４時間培養後、任意の濃度の試料を添加した試験培地に交換し、さらに４８時間培養した。次いで３−(４,５−ジメチル−２−チアゾリル)−２,５−ジフェニルテトラゾリウムブロミド（ＭＴＴ）を４００μｇ／ｍＬ含有する培地に交換して２時間培養し、テトラゾリウム環の開環により生じるフォルマザンを２−プロパノールにて抽出し、マイクロプレートリーダーにて５５０ｎｍの吸光度を測定した。同時に濁度として６５０ｎｍにおける吸光度を測定し、両測定値の差により細胞賦活作用を評価した。評価結果を、試料無添加のブランクにおける細胞賦活作用を１００とした相対値にて表４に示す。なお、表中の**は、ｔ検定における有意確率Ｐ値に対し、有意確率１％未満（Ｐ＜０．０１）を表したものである。
[Evaluation of dermal fibroblast activation by β-endorphin]
Evaluation was performed in the following procedure using human β-endorphin as a sample. Normal human dermal fibroblasts were seeded in a 96-well microplate at 2.0 × 10 4 cells per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% fetal bovine serum. After culturing for 24 hours, the culture medium was replaced with a test medium to which a sample having an arbitrary concentration was added, and further cultured for 48 hours. Next, the medium containing 400 μg / mL of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) was exchanged and cultured for 2 hours. Formazan produced by the opening of the tetrazolium ring was removed. Extraction was performed with 2-propanol, and absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. The evaluation results are shown in Table 4 as relative values with the cell activation effect in the blank with no sample as 100. In addition, ** in the table represents a significance probability of less than 1% (P <0.01) with respect to the significance probability P value in the t test.
表４より明らかなように、β−エンドルフィンを添加した培地において、有意な真皮線維芽細胞賦活作用が認められた。特に、β−エンドルフィンを０．１２５〜０．５ｎｇ／ｍＬ添加した場合には、ブランクと比較して、危険率１％未満で有意な真皮線維芽細胞賦活作用が認められた。このことから、β−エンドルフィンは、優れた真皮線維芽細胞賦活作用を有することが明らかとなった。 As is clear from Table 4, a significant dermal fibroblast activation effect was observed in the medium supplemented with β-endorphin. In particular, when β-endorphin was added in an amount of 0.125 to 0.5 ng / mL, a significant dermal fibroblast activation effect was observed at a risk rate of less than 1% compared to the blank. From this, it was revealed that β-endorphin has an excellent dermal fibroblast activation effect.
［β−エンドルフィンによるチロシナーゼ活性阻害作用の評価］
評価は、試料としてヒトβ−エンドルフィンを用い、以下の手順で行った。正常ヒト表皮メラニン細胞を１ウェル当り３．０×１０４個となるように９６ウェルマイクロプレートに播種した。播種培地にはクラボウ社製Ｍｅｄｉｕｍ１５４Ｓを用いた。２４時間後にＭｅｄｉｕｍ１５４Ｓによって各濃度に調整した試料液に交換し、さらに４８時間培養した。次に、１重量％Ｔｒｉｔｏｎ−Ｘ含有リン酸緩衝液７５μＬに交換し、細胞を完全に溶解させた。その５０μＬを粗酵素液とし、これに基質となる５０μＬの０．０５重量％Ｌ−ドーパ含有リン酸緩衝液を加え、３７℃で２時間静置した。基質添加直後と反応終了時の４０５ｎｍの吸光度をマイクロプレートリーダーにて測定し、生成したドーパメラニン量を測定した。同時に各試料におけるタンパク量を測定し、タンパク量当たりのドーパメラニン生成量を算出した。チロシナーゼ活性阻害作用として、試料無添加のブランクにおけるタンパク量あたりのドーパメラニン生成量を１００とした相対値にて表５に示した。なお、表中の**は、ｔ検定における有意確率Ｐ値に対し、有意確率１％未満（Ｐ＜０．０１）を表したものである。
[Evaluation of Tyrosinase Activity Inhibitory Effect by β-Endorphin]
Evaluation was performed in the following procedure using human β-endorphin as a sample. Normal human epidermal melanocytes were seeded in a 96-well microplate so as to be 3.0 × 10 4 cells per well. As a seeding medium, Medium154S manufactured by Kurabo Industries Co., Ltd. was used. After 24 hours, the medium was replaced with a sample solution adjusted to each concentration by Medium154S, and further cultured for 48 hours. Next, it was replaced with 75 μL of 1 wt% Triton-X-containing phosphate buffer to completely lyse the cells. 50 μL of this was used as a crude enzyme solution, and 50 μL of 0.05 wt% L-dopa-containing phosphate buffer as a substrate was added thereto, and the mixture was allowed to stand at 37 ° C. for 2 hours. The absorbance at 405 nm immediately after the addition of the substrate and at the end of the reaction was measured with a microplate reader, and the amount of produced dopamelanin was measured. At the same time, the amount of protein in each sample was measured, and the amount of dopamelanin produced per amount of protein was calculated. As a tyrosinase activity inhibitory action, it shows in Table 5 by the relative value which set the dopamelanin production amount per protein amount in the blank without a sample to 100. In addition, ** in the table represents a significance probability of less than 1% (P <0.01) with respect to the significance probability P value in the t test.
表５より明らかなように、β−エンドルフィンを添加した培地を用いた場合に、有意なチロシナーゼ活性阻害作用が認められた。特に、β−エンドルフィンを２．５〜３．５ｎｇ／ｍＬ添加した場合には、ブランクと比較して、危険率１％未満で有意なチロシナーゼ活性阻害作用が認められた。このことから、β−エンドルフィンは、チロシナーゼ活性阻害作用を有し、優れた美白作用を発揮することが明らかとなった。 As is clear from Table 5, when a medium supplemented with β-endorphin was used, a significant tyrosinase activity inhibitory effect was observed. In particular, when β-endorphin was added in an amount of 2.5 to 3.5 ng / mL, a significant tyrosinase activity inhibitory effect was observed at a risk rate of less than 1% compared to the blank. From this, it was revealed that β-endorphin has a tyrosinase activity inhibitory action and exhibits an excellent whitening action.
上記の検討により、β−エンドルフィンが皮膚において細胞賦活作用と美白作用を発揮することが明らかとされ、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤を用いてβ−エンドルフィンの産生を促進することにより、β−エンドルフィンの有する細胞賦活作用や美白作用を増強し、しわ，たるみ，しみ，くすみ，乾燥，肌荒れなど皮膚症状の予防や改善が可能であることが明らかとなった。 From the above studies, it has been clarified that β-endorphin exerts cell activation and whitening effects in the skin, and one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant Kelp are effective. Using β-endorphin production promoter as a component to promote the production of β-endorphin enhances cell activation and whitening action of β-endorphin, wrinkles, sagging, blotches, dullness, dryness, rough skin, etc. It was revealed that skin symptoms can be prevented and improved.
最後に、テルミナリア、セイヨウオトギリソウ、トウキンセンカ、ジャイアントケルプより選ばれる１種又は２種以上の抽出物を有効成分とするβ−エンドルフィン産生促進剤を含有する組成物の処方例として、皮膚外用剤及び食品の処方例を示すが、本発明のβ−エンドルフィン産生促進剤を配合する組成物は、これらに限定されるものではない。なお、各処方例に配合したβ−エンドルフィン産生促進剤には、製造例１〜４にて製造した各抽出物を用いた。 Finally, as a prescription example of a composition containing a β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Cinnamon and Giant Kelp as an active ingredient, Although the formulation example of a foodstuff is shown, the composition which mix | blends the beta-endorphin production promoter of this invention is not limited to these. In addition, each extract manufactured by manufacture example 1-4 was used for the beta-endorphin production promoter mix | blended with each prescription example.
［処方例１］クリーム
（１）スクワラン １０．０（重量％）
（２）ステアリン酸 ２．０
（３）水素添加パーム核油 ０．５
（４）水素添加大豆リン脂質 ０．１
（５）セタノール ３．６
（６）親油型モノステアリン酸グリセリン ２．０
（７）グリセリン １０．０
（８）パラオキシ安息香酸メチル ０．１
（９）アルギニン（２０重量％水溶液） １５．０
（10）精製水 ４０．７
（11）カルボキシビニルポリマー（１重量％水溶液） １５．０
（12）β−エンドルフィン産生促進剤 １．０
製法：（１）〜（６）の油相成分を８０℃にて加熱溶解する。一方（７）〜（10）の水相成分を８０℃にて加熱溶解する。これに前記油相成分を攪拌しながら加え、ホモジナイザーにより均一に乳化する。乳化終了後、（11）を加え、冷却を開始し、４０℃にて（12）を加え、均一に混合する。
[Formulation Example 1] Cream (1) Squalane 10.0 (% by weight)
(2) Stearic acid 2.0
(3) Hydrogenated palm kernel oil 0.5
(4) Hydrogenated soybean phospholipid 0.1
(5) Cetanol 3.6
(6) Lipophilic glyceryl monostearate 2.0
(7) Glycerin 10.0
(8) Methyl paraoxybenzoate 0.1
(9) Arginine (20% by weight aqueous solution) 15.0
(10) Purified water 40.7
(11) Carboxyvinyl polymer (1% by weight aqueous solution) 15.0
(12) β-endorphin production promoter 1.0
Production method: The oil phase components (1) to (6) are heated and dissolved at 80 ° C. On the other hand, the aqueous phase components (7) to (10) are dissolved by heating at 80 ° C. The oil phase component is added to this while stirring and uniformly emulsified with a homogenizer. After the emulsification is completed, add (11), start cooling, add (12) at 40 ° C., and mix uniformly.
［処方例２］内服液
（１）β−エンドルフィン産生促進剤 ３．０(重量％)
（２）クエン酸 ０．１
（３）ステビア ０．０１
（４）精製水 ９３．８９
（５）エリスリトール ３．０
製法：（１）〜（５）を均一に混合する。
[Prescription Example 2] Internal liquid (1) β-endorphin production promoter 3.0 (% by weight)
(2) Citric acid 0.1
(3) Stevia 0.01
(4) Purified water 93.89
(5) Erythritol 3.0
Production method: (1) to (5) are mixed uniformly.
Claims (1)
A β-endorphin production promoter containing one or more extracts selected from Terminaria, Hypericum perforatum, Ginseng, Giant kelp.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| JP2009215272A (en) * | 2008-03-06 | 2009-09-24 | Ivy Cosmetics Corp | Skin care preparation for ameliorating roughened skin |
| JP5860577B2 (en) * | 2008-04-17 | 2016-02-16 | 丸善製薬株式会社 | Claudin production promoter and occludin production promoter |
| JP5611755B2 (en) * | 2010-10-18 | 2014-10-22 | 日本メナード化粧品株式会社 | TGF-β inhibitor |
| JP2016006021A (en) * | 2014-06-20 | 2016-01-14 | 株式会社ノエビア | Thioredoxin-related factor expression promoter |
| CN116686990B (en) * | 2023-08-09 | 2023-10-31 | 内蒙古蒙牛乳业（集团）股份有限公司 | Functional composition, functional beverage and preparation method thereof |
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