JP2011001300A - Skin aging inhibitor, inhibitor to frequency decrease of epidermis cell division, and inhibitor to epidermis thickness decrease - Google Patents
Skin aging inhibitor, inhibitor to frequency decrease of epidermis cell division, and inhibitor to epidermis thickness decrease Download PDFInfo
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Abstract
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本発明は、皮膚老化防止剤、表皮細胞***回数減少に対する抑制剤、及び表皮厚減少に対する抑制剤に関する。 The present invention relates to a skin anti-aging agent, an inhibitor for reducing the number of epidermal cell divisions, and an inhibitor for reducing epidermal thickness.
加齢、紫外線曝露等により、皮膚の表皮細胞がダメージを受ける。例えば、表皮細胞数が減少し表皮細胞の代謝が低下して皮膚のターンオーバー速度が遅くなる。その結果、しわの形成、弾性の低下といった皮膚の老化の原因となることが知られている。これらの進行を防止あるいは改善するため、アンチエイジング剤として皮膚老化防止剤が提案されてきた。最近では消費者の商品に対する自然志向及び植物志向を反映して、表皮細胞の代謝促進、賦活化する植物抽出成分等の検索が盛んに行われている。 Skin epithelial cells are damaged by aging, exposure to ultraviolet rays, and the like. For example, the number of epidermal cells decreases, the metabolism of the epidermal cells decreases, and the turnover speed of the skin becomes slow. As a result, it is known to cause skin aging such as formation of wrinkles and decrease in elasticity. In order to prevent or improve these progressions, anti-aging agents for skin have been proposed as anti-aging agents. In recent years, the search for plant extract components and the like that promote and activate epidermal cell metabolism has been actively conducted, reflecting the natural orientation and plant orientation of consumer products.
このような皮膚老化防止剤として、例えば、特許文献1には代謝促進作用を有する皮膚老化防止剤が開示されている。さらに、特許文献2〜3には、細胞賦活作用を有する皮膚老化防止剤が開示されている。 As such a skin antiaging agent, for example, Patent Document 1 discloses a skin antiaging agent having a metabolism promoting action. Furthermore, Patent Documents 2 to 3 disclose skin antiaging agents having a cell activation effect.
本発明は、皮膚の老化を防止できる皮膚老化防止剤の提供を課題とする。また、本発明は、表皮細胞***回数の減少を抑制できる表皮細胞***回数減少に対する抑制剤の提供を課題とする。さらに、本発明は、表皮厚の減少を抑制できる表皮厚減少に対する抑制剤の提供を課題とする。 It is an object of the present invention to provide a skin antiaging agent capable of preventing skin aging. Another object of the present invention is to provide an inhibitor against a decrease in the number of epidermal cell divisions that can suppress a decrease in the number of epidermal cell divisions. Furthermore, this invention makes it a subject to provide the inhibitor with respect to the skin thickness reduction | decrease which can suppress the reduction | decrease of skin thickness.
本発明者等は上記課題に鑑み、鋭意検討を行った。その結果、ニワシロユリ(Lilium candidum)の抽出物が表皮細胞***回数減少、表皮厚減少等の皮膚の老化に対して抑制・防止することができることを見い出した。本発明はこれらの知見に基づいて完成するに至ったものである。 In view of the above problems, the present inventors have conducted intensive studies. As a result, it was found that an extract of Lilium candidum can suppress and prevent skin aging such as a decrease in the number of epidermal cell divisions and a decrease in the thickness of the epidermis. The present invention has been completed based on these findings.
本発明は、ニワシロユリ(Lilium candidum)の抽出物を有効成分として含有する皮膚老化防止剤に関する。
また、本発明は、ニワシロユリ(Lilium candidum)の抽出物を有効成分として含有する表皮細胞***回数減少に対する抑制剤に関する。
さらに、本発明は、ニワシロユリ(Lilium candidum)の抽出物を有効成分として含有する表皮厚減少に対する抑制剤に関する。
The present invention relates to an anti-aging agent for skin containing an extract of Lilium candidum as an active ingredient.
Moreover, this invention relates to the inhibitor with respect to the reduction | decrease in the frequency | count of epidermal cell division containing the extract of a sardine lily ( Lilium candidum ) as an active ingredient.
Furthermore, this invention relates to the inhibitor with respect to the skin thickness reduction | decrease containing the extract of a sardine lily ( Lilium candidum ) as an active ingredient.
本発明の皮膚老化防止剤によれば、効果的に皮膚の老化を防止できる。また、本発明の表皮細胞***回数減少に対する抑制剤によれば、優れた表皮細胞***回数減少抑制作用を有するので、皮膚の老化を防止することができる。さらに、本発明の表皮厚減少に対する抑制剤によれば、優れた表皮厚減少抑制作用を有するので、皮膚の老化を防止することができる。 According to the skin aging inhibitor of the present invention, skin aging can be effectively prevented. Moreover, according to the inhibitor with respect to the reduction | decrease in the number of epidermal cell division of this invention, since it has the outstanding inhibitory effect on the reduction in the number of epidermal cell division, it can prevent skin aging. Furthermore, according to the inhibitor for reducing the skin thickness of the present invention, since it has an excellent inhibitory effect on reducing the skin thickness, aging of the skin can be prevented.
以下、本発明について、その好ましい実施態様に基づき詳細に説明する。
本発明の表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤は、ニワシロユリ(Lilium candidum)の抽出物を有効成分として含有する。
ヒトの表皮は細胞***により絶えず細胞が入れ替わる臓器であり、細胞の状態が皮膚の機能発揮に大きく関わる。しかし、細胞には***限界が存在し、細胞***回数が一定回数を過ぎると、細胞***は停止してしまう。表皮細胞においても***限界が存在し、***停止が表皮老化に関与していることが知られている(例えば、Arch.Gerontol.Geriatr.,1994,Suppl.4,p.185-196;Biochem.Biophys.Res.Comm.,2000,268,p.268-274参照)。また、時間の経過とともに、皮膚の老化現象の1つである表皮の薄化が進む(例えば、J.Dermatol.Sci.,2006,44(3),p.145-152参照)。
前記ニワシロユリの抽出物は、表皮細胞***回数減少、表皮厚減少等の作用を通して皮膚の老化を抑制・防止することができる。前記植物の抽出物を含有する本発明の皮膚老化防止剤、表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤は、優れた表皮細胞***回数減少抑制作用及び表皮厚減少抑制作用を有し、皮膚の老化を防止することができる。
Hereinafter, the present invention will be described in detail based on preferred embodiments thereof.
The inhibitor for reducing the number of epidermal cell divisions and the inhibitor for reducing the epidermal thickness of the present invention contains an extract of sardine lily ( Lilium candidum ) as an active ingredient.
The human epidermis is an organ in which cells are constantly replaced by cell division, and the state of the cells is largely involved in the functioning of the skin. However, cells have division limits, and cell division stops when the number of cell divisions exceeds a certain number. There is also a division limit in epidermal cells, and it is known that mitotic arrest is involved in epidermal aging (see, for example, Arch. Gerontol. Geriatr., 1994, Suppl. 4, p. 185-196; Biochem. Biophys.Res.Com., 2000, 268, p.268-274). In addition, with the passage of time, the epidermis thinning, which is one of the skin aging phenomena, proceeds (for example, see J. Dermatol. Sci., 2006, 44 (3), p.145-152).
The sardine lily extract can suppress / prevent skin aging through actions such as reducing the number of epidermal cell divisions and reducing the thickness of the epidermis. The anti-aging agent for skin according to the present invention containing the plant extract, the inhibitor for reducing the number of epidermal cell divisions, and the inhibitor for reducing the epidermal thickness have an excellent inhibitory effect on reducing the epidermal cell division number and epidermal thickness reduction. In addition, skin aging can be prevented.
ニワシロユリの生理作用については、皮脂分泌抑制作用、アストリンゼント(肌の引きしめ)作用等が知られており(例えば、特表2001−506240号公報等参照)、ニワシロユリの抽出物を含有する皮脂分泌抑制効果を奏する化粧用クレンジング・スキンケア調剤が開示されている(特表2001−506240号公報参照)。しかし、ニワシロユリ抽出物の細胞***回数減少に対する抑制作用、表皮厚減少に対する抑制作用等の皮膚老化防止作用については知られていなかった。 As for the physiological action of sardine lily, sebum secretion inhibitory action, astringent (skin tightening) action, etc. are known (see, for example, JP 2001-506240 A), and sebum secretion inhibitory effect containing an extract of sardine lily A cleansing and skin care preparation for cosmetics is disclosed (see JP-T-2001-506240). However, the anti-skin aging effects such as the inhibitory action on the decrease in the number of cell divisions and the inhibitory action on the reduction in the epidermis thickness of the sardine extract have not been known.
本発明におけるニワシロユリ(Lilium candidum)は、ユリ科(Liliaceae)に属する植物である。バルカン半島及び西アジアを原産とし、別名マドンナリリーともいう。 The sardine lily ( Lilium candidum ) in this invention is a plant which belongs to a lily family ( Liliaceae ). Originating from the Balkans and West Asia, also known as Madonna Lily.
本発明において、前記植物の全ての任意の部分が使用可能である。例えば、上記植物の全木、又は任意の部位(根、根茎、鱗茎、幹、枝、茎、葉、樹皮、樹液、樹脂、花、果実、種子等)、及びそれらの組み合わせのいずれか1つ又は複数を使用することができる。 In the present invention, any arbitrary part of the plant can be used. For example, any one of the whole tree of the above plant, or any part (root, rhizome, bulb, stem, branch, stem, leaf, bark, sap, resin, flower, fruit, seed, etc.) and combinations thereof Or more than one can be used.
本発明において、ニワシロユリの抽出物を得るためには、前記植物の鱗茎を用いるのが好ましい。 In the present invention, in order to obtain an extract of sardine lily, it is preferable to use the bulb of the plant.
本発明において用いる、前記植物の抽出物は、適当な溶媒を用いた常法の抽出方法によって調製することができる。 The plant extract used in the present invention can be prepared by a conventional extraction method using an appropriate solvent.
本発明において、前記植物の抽出物の調製に、上記植物をそのまま、又は乾燥粉砕して用いることもできるが、その水蒸気蒸留物又は圧搾物を用いることもでき、これらは精油等、より精製したものを用いることもでき、また市販品を利用することもできる。上記植物又はその水蒸気蒸留物若しくは圧搾物は、いずれかを単独で、又は2種以上を組み合わせて使用してもよい。 In the present invention, the above plant can be used as it is or after being dried and pulverized for the preparation of the plant extract, but its steam distillate or compressed product can also be used. A thing can also be used and a commercial item can also be utilized. You may use the said plant or its steam distillate or pressing material individually or in combination of 2 or more types.
抽出に用いる溶媒としては、通常植物成分の抽出に用いられるもの、例えば水、石油エーテル、n−ヘキサン、トルエン、クロロホルム、エーテル、酢酸エチル、アセトン、メタノール、エタノール、プロパノール、ブタノール、エチレングリコール、プロピレングリコール、ブチレングリコール等が挙げられ、特に水、エタノール、プロピレングリコール、ブチレングリコールが好ましい。これらは単独で又は2種以上を組み合わせて使用できる。また抽出条件も通常の条件を適用でき、例えば上記植物を0〜100℃で1時間〜30日間浸漬又は加熱還流すればよい。上記植物の抽出物は、そのまま使用できるが、さらに適当な分離手段、例えばゲル濾過、クロマトグラフィー、精密蒸留、活性炭処理等により活性の高い画分を分画して用いることもできる。 Solvents used for extraction are those usually used for extraction of plant components, such as water, petroleum ether, n-hexane, toluene, chloroform, ether, ethyl acetate, acetone, methanol, ethanol, propanol, butanol, ethylene glycol, propylene. Examples include glycol, butylene glycol, and water, ethanol, propylene glycol, and butylene glycol are particularly preferable. These can be used alone or in combination of two or more. Moreover, normal conditions can also be applied as extraction conditions. For example, the plant may be immersed or heated to reflux at 0 to 100 ° C. for 1 hour to 30 days. The plant extract can be used as it is, but it is also possible to fractionate and use a fraction having high activity by an appropriate separation means such as gel filtration, chromatography, precision distillation, activated carbon treatment and the like.
本発明において、前記植物の抽出物はそのまま用いてもよい。あるいは、当該抽出物を希釈、濃縮又は凍結乾燥した後、粉末又はペースト状に調製して用いることもできる。 In the present invention, the plant extract may be used as it is. Alternatively, the extract can be diluted, concentrated, or lyophilized and then prepared into a powder or paste.
本発明において、前記植物の抽出物はそのまま表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤として用いてもよい。あるいは、上記抽出物に、例えば酸化チタン、炭酸カルシウム、蒸留水、乳糖、デンプン等の適当な液体又は固体の賦形剤又は増量剤を加えて用いてもよい。この場合、前記植物の抽出物の量は特に制限されないが、前記抽出物が固形分換算で0.000001〜1重量%含まれるのが好ましく、0.0001〜0.01重量%含まれるのが特に好ましい。 In the present invention, the plant extract may be used as it is as an inhibitor for reducing the number of epidermal cell divisions and an inhibitor for reducing the epidermal thickness. Alternatively, an appropriate liquid or solid excipient or extender such as titanium oxide, calcium carbonate, distilled water, lactose, starch or the like may be added to the above extract. In this case, the amount of the plant extract is not particularly limited, but the extract is preferably contained in an amount of 0.000001 to 1% by weight, particularly preferably 0.0001 to 0.01% by weight in terms of solid content.
本発明の表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤は、表皮細胞***回数減少抑制効果及び表皮厚減少抑制効果を有し、例えば、老化防止用の化粧料や皮膚外用剤等の用途に用いることができる。
本発明の表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤を、上記化粧料や皮膚外用剤に使用する場合、その使用量は、有効成分の含有量により異なるが、例えばクリーム状、軟膏状の場合、皮膚面1cm2当たり1mg〜20mg、液状製剤の場合、同じく1mg〜20mg使用するのが好ましい。
The inhibitor for reducing the number of epidermal cell divisions and the inhibitor for reducing the thickness of the epidermis of the present invention has an effect of suppressing the reduction in the number of epidermal cell divisions and an effect of reducing the thickness of the epidermis. It can be used for
When the inhibitor for reducing the number of epidermal cell divisions and the inhibitor for reducing the skin thickness of the present invention is used in the cosmetic or the external preparation for skin, the amount used varies depending on the content of the active ingredient, for example, cream, In the case of an ointment, it is preferable to use 1 mg to 20 mg per 1 cm 2 of the skin surface, and in the case of a liquid preparation, 1 mg to 20 mg is also used.
本発明の表皮細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤を、老化防止用化粧料や皮膚外用剤に用いる場合、通常の皮膚化粧料等に配合される薬効成分、例えば、ジヒドロキシアセトン、P−MCX、P−1789、微粒子酸化亜鉛、酸化チタン等の紫外線吸収剤、アスコルビン酸等のビタミン類、ヒアルロン酸等の保湿剤、ホルモン剤等を含有させることができる。
また、老化防止用化粧料や皮膚外用剤の形態としては、例えば、クリーム、ローション、乳剤、軟膏、ゲル、パック、フォーム、エッセンス、スティック、パウダー等が挙げられる。
When the inhibitor for reducing the number of epidermal cell divisions and the inhibitor for reducing the skin thickness of the present invention is used for anti-aging cosmetics or skin external preparations, a medicinal component incorporated in normal skin cosmetics, for example, dihydroxyacetone , P-MCX, P-1789, UV absorbers such as fine particle zinc oxide and titanium oxide, vitamins such as ascorbic acid, humectants such as hyaluronic acid, hormone agents, and the like.
Examples of forms of anti-aging cosmetics and skin external preparations include creams, lotions, emulsions, ointments, gels, packs, foams, essences, sticks, and powders.
上記成分のほかに、通常の化粧料又は皮膚外用剤に用いられる各種成分、例えばチョーク、タルク、フラー土、カオリン、デンプン、ゴム、コロイドシリカナトリウムポリアクリレート等の粉体;例えば鉱油、植物油、シリコーン油等の油又は油状物質;例えばソルビタントリオレエート、ソルビタントリステアレート、グリセロールモノオレエート、高分子シリコーン界面活性剤等の乳化剤;パラ−ヒドロキシベンゾエートエステル等の防腐剤;ブチルヒドロキシトルエン等の酸化防止剤;グリセロール、ソルビトール、2−ピロリドン−5−カルボキシレート、ジブチルフタレート、ゼラチン、ポリエチレングリコール等の湿潤剤;トリエタノールアミン又は水酸化ナトリウムのような塩基を伴う乳酸等の緩衝剤;グリセロールエーテル及び合成、動物性又は植物性セラミド等の界面活性剤;密ろう、オゾケライトワックス、パラフィンワックス等のワックス類;増粘剤;活性増強剤;着色料;香料等、を必要に応じ適宜組合せて用いることができる。 In addition to the above components, various components used in normal cosmetics or external preparations for skin, such as chalk, talc, fuller's earth, kaolin, starch, rubber, colloidal silica sodium polyacrylate, etc .; for example, mineral oil, vegetable oil, silicone Oils or oily substances such as oils; for example, emulsifiers such as sorbitan trioleate, sorbitan tristearate, glycerol monooleate, polymeric silicone surfactants; preservatives such as para-hydroxybenzoate esters; antioxidants such as butylhydroxytoluene Agents; humectants such as glycerol, sorbitol, 2-pyrrolidone-5-carboxylate, dibutyl phthalate, gelatin, polyethylene glycol; buffers such as lactic acid with a base such as triethanolamine or sodium hydroxide; glycerol ether A combination of surfactants such as synthetic, animal or plant ceramides; waxes such as beeswax, ozokerite wax, paraffin wax; thickeners; activity enhancers; coloring agents; Can be used.
以下、本発明を実施例に基づきさらに詳細に説明するが、本発明はこれに限定されるものではない。 EXAMPLES Hereinafter, although this invention is demonstrated further in detail based on an Example, this invention is not limited to this.
試験例1 細胞***回数減少抑制試験
正常ヒト新生児表皮角化細胞(商品名:KK−4009、KURABO社製)を、HuMedia−KG増殖添加セット(商品名:KK−6150、KURABO社製)を添加した表皮角化細胞基礎培地(商品名:EpiLife−KG2、KURABO社製)で37℃、5%CO2条件のインキュベーター内で行った。培地は2、3日置きに交換した。
Test Example 1 Cell division frequency reduction suppression test Normal human newborn epidermis keratinocytes (trade name: KK-4009, manufactured by KURABO) and HuMedia-KG proliferation addition set (trade name: KK-6150, manufactured by KURABO) are added. The keratinocyte basal medium (trade name: EpiLife-KG2, manufactured by KURABO) was used in an incubator at 37 ° C. and 5% CO 2 . The medium was changed every few days.
十分数まで増殖させた細胞を、0.4×104個/cm2の濃度となるように25cm2フラスコに播種した。37℃、5%CO2インキュベーターで50〜60%コンフルエントまで培養した後、評価培地に交換し、さらにサブコンフルエントになるまで約1〜2日間培養した。それぞれのフラスコから細胞を回収し、細胞数を計測し、表皮細胞の限界***回数(PDL:Population Doubling Level)を下記の式で算出した。
PDL=(logB−logA)/log2
A:播種した細胞数 B:回収した細胞数
Cells grown to a sufficient number were seeded in a 25 cm 2 flask to a concentration of 0.4 × 10 4 cells / cm 2 . After culturing to 50-60% confluence in a 37 ° C., 5% CO 2 incubator, the culture medium was replaced with an evaluation medium, and further cultured for about 1-2 days until it became subconfluent. Cells were collected from each flask, the number of cells was counted, and the limit number of divisions (PDL: Population Doubling Level) of epidermal cells was calculated by the following formula.
PDL = (logB-logA) / log2
A: Number of cells seeded B: Number of recovered cells
前記評価培地としては、下記に示す組成の培地を用いた。
<本発明>
基礎培地:Epilife
インスリン(10μg/ml)
ハイドロコルチゾン(0.5μg/ml)
抗菌剤(ゲンタマイシン 50μg/ml、アンフォテリシンB 50ng/ml)
0.00003体積%ニワシロユリ抽出物(香栄興業社製)
<比較例>
基礎培地:Epilife
インスリン(10μg/ml)
ハイドロコルチゾン(0.5μg/ml)
抗菌剤(ゲンタマイシン 50μg/ml、アンフォテリシンB 50ng/ml)
As the evaluation medium, a medium having the following composition was used.
<Invention>
Basal medium: Epilife
Insulin (10 μg / ml)
Hydrocortisone (0.5μg / ml)
Antibacterial agent (gentamicin 50 μg / ml, amphotericin B 50 ng / ml)
0.00003 vol% sardine lily extract (manufactured by Koei Kogyo Co., Ltd.)
<Comparative example>
Basal medium: Epilife
Insulin (10 μg / ml)
Hydrocortisone (0.5μg / ml)
Antibacterial agent (gentamicin 50 μg / ml, amphotericin B 50 ng / ml)
回収したそれぞれの細胞を再び0.4×104個/cm2となるように播種し、細胞がコンフルエントになる前のサブコンフルエントの段階で、以下の方法で継代を行った。生理食塩水で細胞を数回洗浄した後、プロナーゼ溶液(極東製薬社製)を加えて37℃で5分間インキュベートすることで、細胞を分散させた。培地を加えて反応停止後、1000rpm、5分間遠心して細胞を回収した。上清を除いて新たな培地で細胞を再懸濁し、細胞数を計測後、一定数を新しい培養フラスコへ播種した。このようにして、細胞***が停止するまで継代培養を繰り返し、各継代数でのPDLを算出した。なお、PDLは継代を重ねるごとの積算値で示した。
ここで用いた培地とは、下記に示す組成の培地である。
基礎培地:Epilife
インスリン(10μg/ml)
ヒト組換え型上皮成長因子(hEGF)(0.1ng/ml)
ハイドロコルチゾン(0.5μg/ml)
ウシ脳下垂体抽出液(BPE)(0.4体積%)
その結果を表1に示す。
Each of the collected cells was seeded again at 0.4 × 10 4 cells / cm 2, and subcultured at the subconfluent stage before the cells became confluent by the following method. After the cells were washed several times with physiological saline, a pronase solution (manufactured by Kyokuto Pharmaceutical Co., Ltd.) was added and incubated at 37 ° C. for 5 minutes to disperse the cells. After stopping the reaction by adding a medium, the cells were collected by centrifugation at 1000 rpm for 5 minutes. The supernatant was removed, the cells were resuspended in a new medium, the number of cells was counted, and a fixed number was seeded in a new culture flask. In this way, subculture was repeated until cell division was stopped, and PDL at each passage number was calculated. In addition, PDL was shown by the integrated value for every passage.
The medium used here is a medium having the composition shown below.
Basal medium: Epilife
Insulin (10 μg / ml)
Human recombinant epidermal growth factor (hEGF) (0.1 ng / ml)
Hydrocortisone (0.5μg / ml)
Bovine pituitary extract (BPE) (0.4% by volume)
The results are shown in Table 1.
表1から明らかなように、ニワシロユリ抽出物を添加した場合、各継代数で表皮細胞のPDLが増加した。この結果から、ニワシロユリ抽出物は、表皮細胞の***が停止するまでの細胞***回数を増加させ表皮細胞の***寿命を延長させる効果、すなわち、表皮細胞の***回数減少抑制効果を有することが示された。 As is apparent from Table 1, when the sardine lily extract was added, the PDL of epidermal cells increased at each passage number. From this result, it is shown that sardine lily extract has the effect of increasing the number of cell divisions until the epidermal cell division stops and extending the division life of the epidermal cells, that is, the effect of suppressing the decrease in the number of divisions of the epidermal cells. It was.
試験例2 表皮厚減少抑制試験
ニワシロユリ抽出物(香栄興業社製)を濃度0.0001体積%となるように添加した培地(商品名:ユリ抽出液、香栄興業株式会社製)で3次元培養皮膚モデルTestskin(商品名、東洋紡社製)を37℃、5%CO2条件のインキュベーター内で6日間培養した。培地は3日目に交換した。なお、比較例としてニワシロユリエキスを添加しない培地を用いて、3次元培養皮膚モデルを培養した。
Test Example 2 Skin Thickness Reduction Inhibition Test Three-dimensional cultured skin with medium (trade name: lily extract, manufactured by Koei Kogyo Co., Ltd.) supplemented with sardine lily extract (manufactured by Koei Kogyo Co., Ltd.) to a concentration of 0.0001% by volume. Model Testskin (trade name, manufactured by Toyobo Co., Ltd.) was cultured in an incubator at 37 ° C. and 5% CO 2 for 6 days. The medium was changed on the third day. As a comparative example, a three-dimensional cultured skin model was cultured using a medium to which no sardine lily extract was added.
培養後の3次元培養皮膚モデルを直径6mmの生検パンチを用いて皮膚片を得た。中性パラホルムアルデヒドで皮膚片を固定しパラフィン切片を作製し、ヘマトキシリン・エオジン染色を行った。
作製したヘマトキシリン・エオジン染色切片を光学顕微鏡で観察し、400倍の倍率で写真撮影し、画像データを取得した。取得した画像データ上で、真皮−表皮の境界線及び表皮−角層の境界線を目視で判別して線を引き、画像解析ソフトImage Pro Plus(商品名、Media Cybernetics社製)で2線間の距離の平均を算出し、表皮厚とした。1つの皮膚モデルについて4視野の写真を撮影して表皮厚を測定し、それらの平均値を算出した。その結果を表2に示す。
Skin pieces were obtained from the cultured three-dimensional cultured skin model using a biopsy punch with a diameter of 6 mm. The skin pieces were fixed with neutral paraformaldehyde, paraffin sections were prepared, and stained with hematoxylin and eosin.
The prepared hematoxylin / eosin-stained section was observed with an optical microscope and photographed at 400 × magnification to obtain image data. On the acquired image data, the boundary line between the dermis and epidermis and the boundary line between the epidermis and the stratum corneum are visually discriminated and drawn, and a line is drawn with image analysis software Image Pro Plus (trade name, manufactured by Media Cybernetics). The average of the distances was calculated as the skin thickness. For one skin model, photographs of 4 fields of view were taken to measure the epidermis thickness, and the average value thereof was calculated. The results are shown in Table 2.
表2から明らかなように、表皮細胞は時間の経過とともに、老化現象の1つである薄化が進む。しかし、ニワシロユリ抽出物を添加することにより、表皮の薄化を抑制することができる。従って、ニワシロユリ抽出物は、表皮厚減少抑制効果を有することが示された。 As is apparent from Table 2, the epidermis cells become thinner as time passes, which is one of the aging phenomena. However, thinning of the epidermis can be suppressed by adding a sardine lily extract. Therefore, it was shown that the sardine lily extract has an effect of suppressing the decrease in the skin thickness.
以上の結果から、ニワシロユリ抽出物は、皮膚の老化現象である表皮細胞***回数減少及び表皮厚減少を抑制することができる。したがって、ニワシロユリ抽出物を有効成分として含有する、本発明の皮膚老化防止剤、細胞***回数減少に対する抑制剤及び表皮厚減少に対する抑制剤は、皮膚の老化を防止できることが示された。 From the above results, the sardine lily extract can suppress the decrease in the number of epidermal cell divisions and the decrease in the epidermal thickness, which are skin aging phenomena. Therefore, it was shown that the skin aging preventive agent of the present invention, the inhibitor for reducing the number of cell divisions and the inhibitor for reducing the epidermis thickness, which contains a scallop extract as an active ingredient, can prevent skin aging.
(処方例)
ニワシロユリ抽出物を有効成分として、下記に示す組成のローション及びクリームを常法により各々調製した。
(Prescription example)
Lotions and creams having the compositions shown below were prepared by conventional methods using sardine lily extract as an active ingredient.
1.ローションの調製
A.に属する成分を溶解し、これとは別にBに属する成分を溶解する。AにBを添加して均一に攪拌混合し、ローションを得た。
A.
(組成) (配合:質量%)
ポリオキシエチレン硬化ひまし油 0.8
エタノール 30.0
B.
(組成) (配合:質量%)
ニワシロユリ抽出物(香栄興業社製) 0.1(乾燥固形分)
ドデシル硫酸ナトリウム 0.12
ドデシルメチルアミンオキシド 0.18
イソプロピルアルコール 15.0
ベンジルアルコール 15.0
グリセリン 2.0
精製水 残量
1. Preparation of lotion A. In addition to this, the component belonging to B is dissolved, and separately, the component belonging to B is dissolved. B was added to A and stirred and mixed uniformly to obtain a lotion.
A.
(Composition) (Composition: Mass%)
Polyoxyethylene hydrogenated castor oil 0.8
Ethanol 30.0
B.
(Composition) (Composition: Mass%)
Sardine lily extract (manufactured by Koei Kogyo Co., Ltd.) 0.1 (dry solid content)
Sodium dodecyl sulfate 0.12
Dodecylmethylamine oxide 0.18
Isopropyl alcohol 15.0
Benzyl alcohol 15.0
Glycerin 2.0
Purified water remaining
2.クリームの調製
A.に属する成分を溶解し、これとは別にBに属する成分を溶解する。AにBを添加して均一に攪拌混合し、乳化後、冷却して、クリームを得た。
A.
(組成) (配合:質量%)
流動パラフィン 10.0
スクワラン 7.0
ホホバ油 3.0
固形パラフィン 3.0
ポリオキシエチレンセチルエーテル 2.0
ソルビタンセスキオレエート 1.0
水酸化カリウム 0.1
B.
(組成) (配合:質量%)
ニワシロユリ抽出物(香栄興業社製) 0.1(乾燥固形分)
グリセリン 3.0
エチルパラベン 0.1
精製水 残量
2. Preparation of cream A. In addition to this, the component belonging to B is dissolved, and separately, the component belonging to B is dissolved. B was added to A and stirred and mixed uniformly. After emulsification, the mixture was cooled to obtain a cream.
A.
(Composition) (Composition: Mass%)
Liquid paraffin 10.0
Squalane 7.0
Jojoba oil 3.0
Solid paraffin 3.0
Polyoxyethylene cetyl ether 2.0
Sorbitan sesquioleate 1.0
Potassium hydroxide 0.1
B.
(Composition) (Composition: Mass%)
Sardine lily extract (manufactured by Koei Kogyo Co., Ltd.) 0.1 (dry solid content)
Glycerin 3.0
Ethylparaben 0.1
Purified water remaining
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JP2013203725A (en) * | 2012-03-29 | 2013-10-07 | Shiseido Co Ltd | Heparan sulphate production promoter |
JP2018184349A (en) * | 2017-04-24 | 2018-11-22 | ポーラ化成工業株式会社 | Eyelid skin surface impression improver and screening method therefor |
CN109481382A (en) * | 2018-12-29 | 2019-03-19 | 株洲千金药业股份有限公司 | A kind of moisture-regulating agent of the water containing bermuda lily and preparation method thereof and facial mask application |
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JP2006111545A (en) * | 2004-10-13 | 2006-04-27 | Nippon Menaade Keshohin Kk | Glutathione reductase activity-enhancing agent |
JP2006232768A (en) * | 2005-02-28 | 2006-09-07 | Nippon Menaade Keshohin Kk | Atp production promoter |
JP2008037812A (en) * | 2006-08-08 | 2008-02-21 | Croda Japan Kk | Cosmetic composition |
JP2008530994A (en) * | 2005-02-22 | 2008-08-14 | フラクソム サイエンシス エイ/エス | Metabolically modified cells for producing resveratrol or oligomeric derivatives or glycoside-linked derivatives thereof |
JP2009161494A (en) * | 2008-01-09 | 2009-07-23 | Noevir Co Ltd | Sirt1 activator |
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JPH0761916A (en) * | 1993-06-30 | 1995-03-07 | Sansho Seiyaku Co Ltd | External agent for skin |
JP2006111545A (en) * | 2004-10-13 | 2006-04-27 | Nippon Menaade Keshohin Kk | Glutathione reductase activity-enhancing agent |
JP2008530994A (en) * | 2005-02-22 | 2008-08-14 | フラクソム サイエンシス エイ/エス | Metabolically modified cells for producing resveratrol or oligomeric derivatives or glycoside-linked derivatives thereof |
JP2006232768A (en) * | 2005-02-28 | 2006-09-07 | Nippon Menaade Keshohin Kk | Atp production promoter |
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JP2013203725A (en) * | 2012-03-29 | 2013-10-07 | Shiseido Co Ltd | Heparan sulphate production promoter |
JP2018184349A (en) * | 2017-04-24 | 2018-11-22 | ポーラ化成工業株式会社 | Eyelid skin surface impression improver and screening method therefor |
CN109481382A (en) * | 2018-12-29 | 2019-03-19 | 株洲千金药业股份有限公司 | A kind of moisture-regulating agent of the water containing bermuda lily and preparation method thereof and facial mask application |
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