JP2013194020A - Antioxidant - Google Patents
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- JP2013194020A JP2013194020A JP2012065104A JP2012065104A JP2013194020A JP 2013194020 A JP2013194020 A JP 2013194020A JP 2012065104 A JP2012065104 A JP 2012065104A JP 2012065104 A JP2012065104 A JP 2012065104A JP 2013194020 A JP2013194020 A JP 2013194020A
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- Cosmetics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
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- Anti-Oxidant Or Stabilizer Compositions (AREA)
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Abstract
Description
本発明は天然物を有効成分とする抗酸化剤に関し、より詳しくは、フリーラジカル捕捉能を有し、安全性が高く、飲食品や外用剤中に配合されて用いられる抗酸化剤に関するものである。 The present invention relates to an antioxidant containing a natural product as an active ingredient, and more particularly to an antioxidant having a free radical scavenging ability, high safety, and blended and used in foods and beverages and external preparations. is there.
近年、特に生体成分を酸化させる要因として、活性酸素が注目されており、その生体への悪影響が問題となっている。活性酸素は、生体細胞内のエネルギー代謝過程で生じるものであり、活性酸素としては、スーパーオキサイド(すなわち酸素分子の一電子還元で生じるスーパーオキシドアニオン:・O2 -)、過酸化水素(H2O2)、ヒドロキシラジカル(・OH)及び一重項酸素(1O2)等が挙げられる。これらの活性酸素は、食細胞の殺菌機構にとって必須であり、ウィルスや癌細胞の除去に重要な働きを果たしている。 In recent years, active oxygen has attracted attention as a factor that particularly oxidizes biological components, and its adverse effect on living organisms has become a problem. Active oxygen is generated in the process of energy metabolism in living cells. Examples of active oxygen include superoxide (that is, superoxide anion generated by one-electron reduction of oxygen molecules: .O 2 − ), hydrogen peroxide (H 2 ). O 2 ), hydroxy radical (.OH), singlet oxygen ( 1 O 2 ) and the like. These active oxygens are essential for the phagocytic sterilization mechanism and play an important role in removing viruses and cancer cells.
しかしながら、活性酸素の過剰な生成は、生体内の膜や組織を構成する生体内分子を攻撃し、各種疾患を誘発する。通常、生体内で生産され、他の活性酸素の出発物質ともなっているスーパーオキサイドは、細胞内に含まれているスーパーオキサイドジスムターゼ(SOD)の触媒作用により逐次消去されているが、スーパーオキサイドの産生が過剰である場合、又はSODの作用が低下している場合には、スーパーオキサイドの消去が不十分となり、スーパーオキサイド濃度が高くなり、これが関節リウマチやベーチェット病等の組織障害、心筋梗塞、脳卒中、白内障、しわ、糖尿病、動脈硬化、肩凝り、冷え性等を引き起こす。 However, excessive production of active oxygen attacks in vivo molecules constituting membranes and tissues in the living body and induces various diseases. Normally, superoxide produced in vivo and used as a starting material for other active oxygen is sequentially eliminated by the catalytic action of superoxide dismutase (SOD) contained in the cells. Is excessive or when the SOD action is reduced, superoxide elimination is insufficient, and the superoxide concentration increases, which may cause tissue damage such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke Cause cataracts, wrinkles, diabetes, arteriosclerosis, stiff shoulders, coldness.
特に皮膚は、酸素や紫外線等の外界からの刺激も直接受けることから、活性酸素による酸化ストレスのダメージが大きな組織である。近年では、紫外線により発生した種々の活性酸素が、皮脂や脂質の過酸化、蛋白変性、酵素阻害等を引き起こし、それが、短期的には皮膚の炎症などを誘発する。また、長期的には、老化やガンなどの原因となると考えられている。さらに活性酸素や過酸化脂質は、アトピー性皮膚炎や接触皮膚炎、乾癬などの皮膚疾患にも関与すると考えられている。このように、皮膚老化や皮膚疾患には、活性酸素(フリーラジカル)が深く関与している。 In particular, the skin is directly affected by external stimuli such as oxygen and ultraviolet rays, and is a tissue that is greatly damaged by oxidative stress due to active oxygen. In recent years, various active oxygens generated by ultraviolet rays cause sebum and lipid peroxidation, protein denaturation, enzyme inhibition, etc., which in the short term induce skin inflammation and the like. In the long term, it is thought to cause aging and cancer. Furthermore, active oxygen and lipid peroxide are considered to be involved in skin diseases such as atopic dermatitis, contact dermatitis, and psoriasis. Thus, active oxygen (free radical) is deeply involved in skin aging and skin diseases.
したがって、活性酸素や生体内ラジカルの生成を阻害・抑制することにより、しわ形成や弾力低下等の皮膚の老化や、関節リウマチやベーチェット病等の組織障害、心筋梗塞、脳卒中、白内障、糖尿病、動脈硬化、肩凝り、冷え性等の活性酸素が関与する各種障害を予防、治療又は改善できるものと考えられる。 Therefore, by inhibiting and suppressing the generation of active oxygen and in vivo radicals, skin aging such as wrinkle formation and reduced elasticity, tissue disorders such as rheumatoid arthritis and Behcet's disease, myocardial infarction, stroke, cataracts, diabetes, arteries It is considered that various disorders involving active oxygen such as hardening, stiff shoulders, and coldness can be prevented, treated or improved.
上記のような活性酸素による酸化を防止するため、従来様々な抗酸化剤が開発されている。例えば、酸化防止剤として知られているビタミンEやビタミンCは、生体内におけるフリーラジカル捕捉型抗酸化物質である。また、BHTやBHAの合成抗酸化物質も知られている。また、植物由来の酸化防止剤としては、シイタケ、エノキタケ、シメジ、カワラタケ、マツタケ、マンネンタケ、ホウウロクタケ、ナメコ、その他の担子菌類の抽出物が報告されている(特許文献1〜3)。さらに、ゴマノハグサ科モウズイカ属植物の抽出物からなる抗酸化剤や(特許文献4)、ムラサキ科カキバチシャノキ属植物の抽出物からなる抗酸化剤(特許文献5)、松樹皮抽出物とセサミンからなる抗酸化剤(特許文献6)、サガリバナ、ナンヨウザクラ、キントラノオ等の抽出物からなる抗酸化剤(特許文献7)、羅漢果花部の抽出物からなる抗酸化剤(特許文献8)などが報告されている。
In order to prevent oxidation due to active oxygen as described above, various antioxidants have been conventionally developed. For example, vitamin E and vitamin C known as antioxidants are free radical scavenging antioxidants in vivo. In addition, synthetic antioxidants such as BHT and BHA are also known. In addition, as plant-derived antioxidants, extracts of shiitake, enokitake, shimeji, kawatake, matsutake, garlic mushroom, spinach, nameko, and other basidiomycetes have been reported (
このように、植物由来の抗酸化剤については、新しい植物で新たな抗酸化力を奏する植物を見出すことが求められている。そこで本発明者らはこのような従来の事情に対処して、新しい植物由来の抗酸化剤を提供することを目的とする。 Thus, for plant-derived antioxidants, it is required to find new plants that exhibit new antioxidant power. Therefore, the present inventors have dealt with such a conventional situation and an object thereof is to provide a new plant-derived antioxidant.
本発明者等は、このような現状に鑑み、鋭意研究を重ねた結果、特定の植物抽出物に優れた抗酸化作用があることを見出し、本発明を完成するに至った。 As a result of intensive studies in view of such a current situation, the present inventors have found that a specific plant extract has an excellent antioxidant action, and have completed the present invention.
本発明は、(a)マメ科シカクマメ(Psophocarpus)属に属する植物の中から選ばれる1種または2種以上の植物の抽出物を有効成分とする抗酸化剤である。 The present invention is (a) an antioxidant comprising as an active ingredient an extract of one or more plants selected from plants belonging to the genus Psophocarpus.
また本発明は、(a)マメ科シカクマメ(Psophocarpus)属に属する植物の中から選ばれる1種または2種以上の植物の抽出物および(b)ローヤルゼリーを有効成分とする抗酸化剤である。 Further, the present invention is (a) an extract of one or more plants selected from plants belonging to the genus Psophocarpus, and (b) an antioxidant containing royal jelly as an active ingredient.
本発明の抗酸化剤は、フリーラジカル捕捉能を有し、優れた抗酸化作用を有すると共に、天然物由来成分であるから安全性が高く、経口摂取、経皮投与のいずれも可能である。 The antioxidant of the present invention has a free radical scavenging ability, has an excellent antioxidant action, and is a natural product-derived component, so it is highly safe and can be taken orally or transdermally.
本発明で用いられる(a)マメ科シカクマメ(Psophocarpus)属に属する植物は、マメ科の熱帯産つる性の草本である。本発明ではシカクマメ属に属する植物であれば特に限定されるものではなく、任意に用いることができる。本発明では中でも、シカクマメ(学名:Psophocarpus tetragonolobus)が好ましく用いられる。改良品種として「ウリズン」等が知られる。市販品としては、その種子をサカタのタネなどで購入できる。
シカクマメ属植物抽出物は、従来、ラミニン5産生促進作用や細胞増殖促進作用等を有する成分としては知られているが(特開2003−313135号公報、特開2010−24211号公報参照)、抗酸化効果については知られておらず、本発明者が初めて見出したものである。
The plant belonging to the genus (a) leguminous winged bean (Psophocarpus) used in the present invention is a herbaceous tropical vine. In this invention, if it is a plant which belongs to the winged genus, it will not specifically limit, It can use arbitrarily. In the present invention, winged bean (scientific name: Psophocarpus tetragonolobus) is preferably used. “Urisun” is known as an improved variety. As a commercial product, the seeds can be purchased from Sakata Seeds.
The winged plant extract is conventionally known as a component having a laminin 5 production promoting action, a cell growth promoting action or the like (see Japanese Patent Application Laid-Open Nos. 2003-313135 and 2010-24211). The oxidation effect is not known and has been found for the first time by the present inventors.
本発明に用いられるシカクマメ(Psophocarpus)属植物の抽出物は、シカクマメ(Psophocarpus)属植物の葉、茎、枝、花、樹皮、種子、さや、根茎または植物全草等を抽出溶媒と共に浸漬または加熱還流した後、濾過し濃縮して得られる。使用部位としては、植物体の任意の部位を用いることができるが、種子を用いるのが特に好ましい。 The extract of the plant belonging to the genus Psophocarpus used in the present invention is obtained by immersing or heating leaves, stems, branches, flowers, bark, seeds, pods, rhizomes or whole plants of the plant of the genus Psophocarpus together with an extraction solvent. After refluxing, it is obtained by filtration and concentration. Although any part of the plant can be used as the part to be used, it is particularly preferable to use seeds.
シカクマメ(Psophocarpus)属植物の抽出物は、常法により得ることができ、例えばシカクマメ(Psophocarpus)属植物を抽出溶媒とともに浸漬または加熱還流した後、濾過し、濃縮して得ることができる。抽出溶媒としては、通常抽出に用いられる溶媒であれば任意に用いることができ、例えば、水、メタノール、エタノール、プロピレングリコール、1,3−ブチレングリコール、グリセリン等のアルコール類、含水アルコール類、クロロホルム、ジクロルエタン、四塩化炭素、アセトン、酢酸エチル、ヘキサン等の有機溶媒等を、それぞれ単独あるいは組み合わせて用いることができる。上記溶媒で抽出して得た抽出液をそのまま、あるいは濃縮したエキスを吸着法、例えばイオン交換樹脂を用いて不純物を除去したものや、ポーラスポリマー(例えばアンバーライトXAD−2)のカラムにて吸着させた後、メタノールまたはエタノールで溶出し、濃縮したものも使用することができる。また分配法、例えば水/酢酸エチルで抽出した抽出物等も用いられる。 An extract of a genus Psophocarpus can be obtained by a conventional method. For example, a genus Psophocarpus can be obtained by dipping or heating under reflux with an extraction solvent, followed by filtration and concentration. As the extraction solvent, any solvent that is usually used for extraction can be used. For example, water, methanol, ethanol, propylene glycol, 1,3-butylene glycol, glycerin and other alcohols, hydrous alcohols, chloroform , Organic solvents such as dichloroethane, carbon tetrachloride, acetone, ethyl acetate, hexane and the like can be used alone or in combination. The extract obtained by extraction with the above solvent is used as it is, or the concentrated extract is adsorbed by an adsorption method, for example, by removing impurities using an ion exchange resin, or by a porous polymer (for example, Amberlite XAD-2) column. Then, it can be used after elution with methanol or ethanol. Further, a partitioning method, for example, an extract extracted with water / ethyl acetate can be used.
こうして得られる抽出液は、そのまま、あるいはエタノール等でさらに希釈し、または固化後、乾燥物をそのまま、もしくは乾燥物を例えばエタノールに再溶解して、抗酸化剤として用いることができる。 The extract thus obtained can be used as an antioxidant as it is or after further dilution with ethanol or the like, or after solidification, and the dried product can be used as it is, or the dried product can be redissolved in, for example, ethanol.
(ローヤルゼリー)
ローヤルゼリーは、ミツバチの働き蜂から女王蜂に与えられる特別食である。ミツバチの下咽頭腺、大顎腺から分泌される乳白色糊状の物質で、主な成分はタンパク質、炭水化物、脂肪、無機物、ビタミン等である。
本発明のローヤルゼリーとしては、女王蜂を育てるために蜜蜂が分泌するものを収集したものを用いることもでき、また一般に市販されているものを用いることもできる。本発明のローヤルゼリーとしては、ローヤルゼリーをそのまま用いても良く、或いは、ローヤルゼリーを適宜前処理して得た乾燥物を用いても良い。
本発明で用いられるローヤルゼリーは、蜜蜂の若い働き蜂の頭部にある分泌腺から分泌される乳白色の粘液性物質であるローヤルゼリーを、アルコール、多価アルコール、その他各種有機溶媒、水、水性アルコール等の抽出溶媒で抽出したものを使用することが好ましい。
(Royal jelly)
Royal jelly is a special meal given to queen bees by bee workers. It is a milky white paste-like substance secreted from the hypopharyngeal gland and the maxillary gland of bees. The main ingredients are proteins, carbohydrates, fats, minerals, vitamins and so on.
As the royal jelly of the present invention, those collected by bees secreted for raising queen bees can be used, and those generally available on the market can also be used. As the royal jelly of the present invention, the royal jelly may be used as it is, or a dried product obtained by appropriately pretreating the royal jelly may be used.
The royal jelly used in the present invention is a milky white mucous substance secreted from a secretory gland in the head of a young worker bee, such as alcohol, polyhydric alcohol, other various organic solvents, water, aqueous alcohol, etc. It is preferable to use one extracted with an extraction solvent.
(b)ローヤルゼリーについては、高麗人参等と併用して肌荒れ防止効果を奏すること(特開平9−286709号公報)が知られている。また、ローヤルゼリー由来物が抗酸化活性を有することも知られている(特開2005−263782号公報、特開2009−274960号公報等)。
しかし本発明者らの実験によれば、ローヤルゼリー単独で抗酸化活性は認められず、シカクマメ属植物とローヤルゼリーとを併用することではじめて優れた抗酸化活性を有するという結果が得られている。
(B) As for royal jelly, it has been known that the effect of preventing rough skin is obtained in combination with ginseng (JP-A-9-286709). It is also known that royal jelly-derived products have antioxidant activity (Japanese Patent Application Laid-Open No. 2005-263788, Japanese Patent Application Laid-Open No. 2009-274960, etc.).
However, according to experiments by the present inventors, royal jelly alone has no antioxidant activity, and it has been shown that it has excellent antioxidant activity only when a winged genus plant and royal jelly are used in combination.
本発明の抗酸化剤は(a)マメ科シカクマメ(Psophocarpus)属に属する植物の中から選ばれる1種または2種以上の植物の抽出物、あるいは該抽出物と、(b)ローヤルゼリーとからなることが好ましいが、本発明の効果を損なわない範囲において他の種々の成分を含有することが出来る。 The antioxidant of the present invention comprises (a) an extract of one or more plants selected from plants belonging to the genus Psophocarpus, or the extract, and (b) royal jelly. Although it is preferable, other various components can be contained within a range not impairing the effects of the present invention.
また本発明の抗酸化剤は(a)マメ科シカクマメ(Psophocarpus)属に属する植物の中から選ばれる1種または2種以上の植物の抽出物と、(b)ローヤルゼリーとを含むことで、抗酸化効果は一層高められる。 The antioxidant of the present invention contains (a) an extract of one or more plants selected from plants belonging to the genus Psophocarpus, and (b) a royal jelly. The oxidation effect is further enhanced.
本発明の抗酸化剤において、(a)マメ科シカクマメ(Psophocarpus)属植物抽出物と、(b)ローヤルゼリーを含む場合、その配合比率は、1:10〜80:1の割合(乾燥質量での質量比)で組合せて配合することが好ましい。この範囲内であると抗酸化効果が特に優れる。 In the antioxidant of the present invention, when (a) a leguminous winged bean (Psophocarpus) plant extract and (b) royal jelly are included, the blending ratio is 1:10 to 80: 1 (in dry mass) (Mass ratio) is preferably combined. Within this range, the antioxidant effect is particularly excellent.
本発明に記載の抗酸化剤は、そのまま、または種々の成分を加えて、飲食品類、化粧品類として経口または経皮で用いることができる。 The antioxidants described in the present invention can be used orally or percutaneously as they are or as various foods and cosmetics with various components added.
飲食品類としては、本発明に記載の抗酸化剤をそのまま、または種々の栄養成分を加えて、食用に適した形態、例えば、粉末状・粒状・顆粒状・液状・ペースト状・クリーム状・タブレット状・カプセル状・カプレット状・ソフトカプセル状・錠剤状・棒状・板状・ブロック状・丸薬状・固形状・ゲル状・ゼリー状・グミ状・ウエハース状・ビスケット状・飴状・チュアブル状・シロップ状・スティック状などに成形して食品素材として提供することができる。また、水、牛乳、豆乳、果汁飲料、乳清飲料、清涼飲料、青汁、ヨーグルトなどに添加して使用してもよい。 As foods and beverages, the antioxidant described in the present invention is used as it is or various nutritional components are added, and the food is suitable, for example, powder, granule, granule, liquid, paste, cream, tablet , Capsule, caplet, soft capsule, tablet, rod, plate, block, pill, solid, gel, jelly, gummy, wafer, biscuit, bowl, chewable, syrup It can be formed into a shape or stick shape and provided as a food material. Further, it may be used by adding to water, milk, soy milk, fruit juice drink, whey drink, soft drink, green juice, yogurt and the like.
化粧品類としては、例えば、化粧水、化粧クリーム、乳液、クリーム、パック、ヘアト
ニック、ヘアクリーム、シャンプー、ヘアリンス、トリートメント、ボディシャンプー、
洗顔剤、石鹸、ファンデーション、整髪料などの形態で化粧品素材として使用できる。
As cosmetics, for example, lotion, cosmetic cream, milky lotion, cream, pack, hair tonic, hair cream, shampoo, hair rinse, treatment, body shampoo,
It can be used as a cosmetic material in the form of facial cleansers, soaps, foundations, hair styling, etc.
本発明の抗酸化剤を食品に配合して用いる場合、本発明の抗酸化剤の配合量(乾燥質量)は、それらの種類、目的、形態、利用方法などに応じて、適宜決めることができ、例えば、食品全量中に0.1〜90質量%程度とすることができる。特に、保健用飲食品等として利用する場合には、本発明の有効成分を所定の効果が充分発揮されるような量で含有させることが好ましい。 When the antioxidant of the present invention is used in a food, the amount (dry mass) of the antioxidant of the present invention can be appropriately determined according to the type, purpose, form, method of use and the like. For example, about 0.1-90 mass% can be made into food whole quantity. In particular, when used as a health food or drink, it is preferable to contain the active ingredient of the present invention in such an amount that a predetermined effect is sufficiently exhibited.
また、本発明の抗酸化剤は、大人では(a)マメ科シカクマメ(Psophocarpus)属に属する植物の中から選ばれる1種または2種以上の植物の抽出物を1日当たり50mg以上、好ましくは50mg〜1000mgを投与することが好ましい。また、(b)ローヤルゼリーは、1日当たり生ローヤルゼリーとして10mg以上、好ましくは50mg〜2000mgを投与することが好ましい。1日当たりの投与量が少なすぎると効果が十分に発揮されない。1日当たりの投与量の上限は本発明の効果を損なわない範囲において特に限定されないが、過剰に配合しても増量に見合った顕著な効果が得られない。 In addition, the antioxidant of the present invention is 50 mg or more, preferably 50 mg, of an extract of one or more kinds of plants selected from among the plants belonging to the genus (a) leguminous family (Psophocarpus) in adults. It is preferred to administer ~ 1000 mg. In addition, (b) Royal jelly is preferably administered in an amount of 10 mg or more, preferably 50 mg to 2000 mg as raw royal jelly per day. If the dose per day is too small, the effect is not sufficiently exhibited. The upper limit of the dose per day is not particularly limited as long as the effect of the present invention is not impaired, but a remarkable effect commensurate with the increase in dosage cannot be obtained even if it is added in excess.
経皮吸収の形態としては、例えば、皮膚化粧料とすることができる。特に制限はなく、各種用途から適宜選択することができ、例えば、軟膏、クリーム、乳液、ローション、パック、ゼリー、リップクリーム、口紅、入浴剤、アストリンゼント、などが挙げられる。 As a form of transdermal absorption, for example, a skin cosmetic can be used. There is no restriction | limiting in particular and it can select suitably from various uses, For example, ointment, cream, emulsion, lotion, pack, jelly, lip balm, lipstick, bath agent, astringent, etc. are mentioned.
本発明による抗酸化剤の前記皮膚化粧料全体に対する配合量は、皮膚化粧料の種類や抽出物の生理活性等によって適宜調整することができるが、乾燥残分として0.0001〜10質量%が好ましく、0.001〜1質量%がより好ましい。 The blending amount of the antioxidant according to the present invention with respect to the entire skin cosmetic can be adjusted as appropriate depending on the type of skin cosmetic, the physiological activity of the extract, etc., but the dry residue is 0.0001 to 10% by mass. Preferably, 0.001-1 mass% is more preferable.
必要に応じて本発明の効果を損なわない範囲で、その皮膚化粧料の製造に通常使用される各種主剤及び助剤、その他成分を使用することができる。
その他の成分としては、抗酸化作用の妨げにならない限り、特に制限はなく、目的に応じて適宜選択した成分が挙げられ、例えば、収斂剤、殺菌剤、抗菌剤、紫外線吸収剤、細胞賦活剤、油脂類、ロウ類、炭化水素類、脂肪酸類、アルコール類、エステル類、界面活性剤、香料、などが挙げられる。これらの成分は、本体重減少化剤と共に併用した場合、相乗的に作用して、通常期待される以上の優れた作用効果をもたらすことがある。
If necessary, various main ingredients and auxiliaries usually used in the production of the skin cosmetics and other components can be used as long as the effects of the present invention are not impaired.
The other components are not particularly limited as long as they do not hinder the antioxidant action, and include components appropriately selected according to the purpose. Examples include astringents, bactericides, antibacterial agents, ultraviolet absorbers, cell activators. , Oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, fragrances, and the like. These components, when used in combination with the weight loss agent, may act synergistically to provide superior effects than would normally be expected.
本発明について以下に実施例を挙げてさらに詳述するが、本発明はこれによりなんら限定されるものではない。配合量は特記しない限り質量%で示す。 The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto. Unless otherwise specified, the amount is shown in mass%.
試験例1〜4
A.試料の調製
(1)シカクマメ抽出物溶液の調製
シカクマメの種子50gを、室温で1週間90%エタノールに浸漬し、抽出液を濃縮し、シカクマメ90%エタノール抽出物(乾燥物)2.9gを得た。
得られたシカクマメ抽出物をジメチルスルホキシド(DMSO)mlに溶解し、この溶液を用いて実分の終濃度が0.005%(ローヤルゼリー溶液と併用する場合),0.01%(ローヤルゼリー溶液と併用しない場合)となるように培地中に添加した。
Test Examples 1-4
A. Preparation of sample (1) Preparation of winged bean extract 50 wings of winged bean seeds were immersed in 90% ethanol for 1 week at room temperature, and the extract was concentrated to obtain 2.9 g of winged bean 90% ethanol extract (dried). It was.
The obtained winged bean extract is dissolved in dimethyl sulfoxide (DMSO) ml, and using this solution, the final concentration of the actual component is 0.005% (when used in combination with royal jelly solution), 0.01% (used in combination with royal jelly solution) If not, it was added to the medium.
(2)ローヤルゼリー溶液の調製
ローヤルゼリー50gに85%エタノール溶液500mLを加え、10分間攪拌し、1時間超音波処理を行った。不溶成分を限外ろ過膜によりろ過してローヤルゼリー溶液を得た(生ローヤルゼリー換算20%、乾燥残分7%)。
得られた溶液を培地に溶解して実分終濃度0.005%(シカクマメ抽出物と併用する場合),0.01%(シカクマメ抽出物と併用しない場合)となるように添加した。
(2) Preparation of Royal Jelly Solution 500 mL of 85% ethanol solution was added to 50 g of royal jelly, stirred for 10 minutes, and subjected to ultrasonic treatment for 1 hour. The insoluble component was filtered through an ultrafiltration membrane to obtain a royal jelly solution (raw royal jelly equivalent 20%, dry residue 7%).
The obtained solution was dissolved in a medium and added so as to have a final concentration of 0.005% (when used together with winged bean extract) and 0.01% (when not used together with winged bean extract).
B.抗酸化活性(フリーラジカル捕捉活性)の測定
ヒト表皮角化細胞を、表皮角化細胞専用培地(KGM)にて1×105cells/mlの濃度で24穴プレートに播種した。細胞接着後、0.1%BSA含有基礎培地(KBM)に薬剤(試験試料)を添加したものに置換し一晩放置した。
翌日水溶性ラジカル開始剤AAPH(2,2'-Azobis-(2-Amidinopropane)Dihydrochloride)を80mM添加した0.1%BSA含有基礎培地(KBM)に置換し、3.5時間後、それぞれの細胞生存率についてラジカル開始剤を添加していないものを100とした時の値で比較した。
B. Measurement of Antioxidant Activity (Free Radical Scavenging Activity) Human epidermal keratinocytes were seeded in a 24-well plate at a concentration of 1 × 10 5 cells / ml in an epidermis keratinocyte-dedicated medium (KGM). After the cell adhesion, the basal medium (KBM) containing 0.1% BSA was replaced with a drug (test sample) added and left overnight.
The next day, the basal medium (KBM) containing 0.1% BSA supplemented with 80 mM of water-soluble radical initiator AAPH (2,2'-Azobis- (2-Amidinopropane) dihydrochloride) was replaced. The survival rate was compared with the value when 100 was not added the radical initiator.
C.結果
結果を表1および図1に示す。
C. Results The results are shown in Table 1 and FIG.
図1は、表1の結果をグラフにしたものである。
上記結果から分かるように、薬剤を何も添加しない場合はラジカル開始剤の添加により細胞生存率は47.1%に低下した(試験例1)。シカクマメ抽出物を0.01%添加するとその低下が有意に抑制された(試験例2)。シカクマメ抽出物とローヤルゼリー溶液を共に(各0.005%)添加した場合、さらに生存率は高まった(試験例3)。
一方、ラジカル開始剤存在下でローヤルゼリー溶液0.01%のみを加えても、ラジカル開始剤による生存率の低下を抑制することはできなかった(試験例4)。
FIG. 1 is a graph of the results in Table 1.
As can be seen from the above results, when no drug was added, the cell viability decreased to 47.1% by the addition of the radical initiator (Test Example 1). When 0.01% winged bean extract was added, the decrease was significantly suppressed (Test Example 2). When both winged bean extract and royal jelly solution were added (0.005% each), the survival rate further increased (Test Example 3).
On the other hand, even when only 0.01% of the royal jelly solution was added in the presence of the radical initiator, the decrease in the survival rate due to the radical initiator could not be suppressed (Test Example 4).
これらの結果から、シカクマメエキスの添加はラジカル開始剤による生存率低下を抑制し、さらにローヤルゼリーを加えるとその効果はさらに高まることが明らかとなった。 From these results, it was clarified that the addition of winged bean extract suppressed the decrease in the survival rate due to the radical initiator, and the effect was further enhanced by adding royal jelly.
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