JP3553477B2 - Active oxygen scavenging enzyme activity promoter - Google Patents
Active oxygen scavenging enzyme activity promoter Download PDFInfo
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- JP3553477B2 JP3553477B2 JP2000272718A JP2000272718A JP3553477B2 JP 3553477 B2 JP3553477 B2 JP 3553477B2 JP 2000272718 A JP2000272718 A JP 2000272718A JP 2000272718 A JP2000272718 A JP 2000272718A JP 3553477 B2 JP3553477 B2 JP 3553477B2
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Description
【０００１】
【発明の属する技術分野】
本発明は、活性酸素消去酵素の活性促進剤に関し、特に紫外線や加齢などにより低下した活性酸素消去酵素の活性を促進することにより皮膚をはじめとする生体の酸化予防に有効な活性酸素消去酵素の活性促進剤に関する。
【０００２】
【従来の技術】
皮膚は生体の最外層に位置し、紫外線等の影響により活性酸素が発生しやすい臓器であり、絶えずその酸素ストレスに曝されている。一方、皮膚細胞内には活性酸素消去酵素が存在しており、その能力を超える活性酸素が発生しないかぎり活性酸素の傷害から皮膚細胞を防衛している。ところが、皮膚細胞内活性酸素消去酵素の活性は加齢とともに低下することが知られており、活性酸素による傷害がその防御反応を凌駕したとき、皮膚は酸化され、細胞機能が劣化して老化してゆくと考えられる。そこで、活性酸素による傷害からの防御を目的として特開平９−１１８６３０や特開平９−２０８４８４などのように活性酸素消去剤や抗酸化剤が検討され、ＳＯＤやカタラーゼ等の活性酸素消去酵素、ＳＯＤ様活性物質などの活性酸素消去剤や抗酸化剤を配合した食品、化粧品、医薬部外品または医薬品等が開発されている。
【０００３】
【発明が解決しようとする課題】
しかしながら、ＳＯＤやカタラーゼ等の活性酸素消去酵素は安定性に問題があり、活性酸素消去剤や抗酸化剤は、その効果が充分でないものが多いため、安定性に優れ、効果の高い活性酸素消去酵素の活性促進剤が求められている。
【０００４】
【課題を解決するための手段】
このような事情により、本発明者らは活性酸素による傷害からの防御作用に優れた活性酸素消去酵素の活性促進剤を得るために鋭意検討した結果、メハジキ、カンゾウ、ジオウ、ロクジョウの抽出物が、安定かつ皮膚細胞が本来持っている活性酸素消去酵素の活性を高めることを見出し、本発明を完成するに至った。
【０００５】
すなわち、本発明は、メハジキ、カンゾウ、ジオウ、ロクジョウの抽出物を含有する活性酸素消去酵素の活性促進剤である。
【０００６】
本発明で用いられるメハジキ（学名：Ｌｅｏｎｕｒｕｓ ｓｉｂｉｒｉｃｕｓ Ｌ．）は、シソ科メハジキ属に属する草本で、本州、四国、九州、朝鮮及び中国から東南アジアにかけて山野の日当りの良い場所に自生する。メハジキの地上部の全草は益母草（ヤクモソウ）の名称で、生薬として産後の止血、月経不順、めまい及び腹痛等に用いられている。
【０００７】
本発明で用いられるカンゾウ（学名：Ｇｌｙｃｙｒｒｈｉｚａ ｕｒａｌｅｎｓｉｓ）は、マメ科カンゾウ属に属する草本で、アフリカ、ヨーロッパからアジア、オーストラリア、北米西部に分布する。根や走出茎（ストロン）が漢方薬で甘草と呼ばれ利用されている。近縁のスペインカンゾウ（学名：Ｇｌｙｃｙｒｒｈｉｚａ ｇｌａｂｒａ）を用いてもよい。
【０００９】
本発明で用いられるジオウ（学名：Ｒｅｈｍａｎｎｉａ ｇｌｕｔｉｎｏｓａ Ｌｉｂｏｓｃｈｉｔｚ）は、ゴマノハグサ科ジオウ属に属する草本で、中国中北部、朝鮮に分布する。ジオウの根を乾燥したものは地黄と呼ばれ生薬として用いられる。
【００１０】
本発明で用いられるロクジョウは、シカ科のマンシュウアカジカ（学名：Ｃｅｒｖｕｓ （Ｃｅｒｖｕｓ） ｅｌａｐｈｕｓ Ｌ． ｖａｒ． ｘａｎｔｈｏｐｙｇｕｓ Ｍｉｌｎｅ−Ｅｄｗａｒｄｓ）およびマンシュウジカ（学名： Ｃｅｒｖｕｓ （Ｓｉｋａ） ｎｉｐｐｏｎ Ｔｅｍｍｉｎｃｋ ｖａｒ． ｍａｎｔｃｈｕｒｉｃｕｓ Ｓｗｉｎｈｏｅ）の雄のまだ角化していない、もしくはわずかに角化した幼角（袋角）である。
【００１１】
本発明で使用するメハジキの抽出物とは、メハジキの葉、茎、樹皮、花、実、根等の植物体の一部又は全草から抽出したものである。好ましくは、生薬として用いられるメハジキの地上部の全草から抽出して得られるものが良い。カンゾウの抽出物とは、カンゾウの葉、茎、樹皮、花、実、根等の植物体の一部又は全草から抽出したものである。好ましくは、生薬として用いられるカンゾウの根およびストロンを乾燥したものから抽出したものが良い。ジオウの抽出物とは、ジオウの葉、茎、樹皮、花、実、根等の植物体の一部又は全草から抽出したものである。好ましくは、生薬として用いられるジオウの根を乾燥したものが良い。ロクジョウの抽出物とは、マンシュウアカジカおよびマンシュウジカの袋角から抽出したものである。その調製方法は特に限定されず、例えば、加熱抽出したものであっても良いし、常温抽出したものであっても良い。また、上記抽出物の中には市販されているものもあり、これらを用いることができる。
【００１２】
抽出する溶媒としては、例えば、水、低級１価アルコール類（メタノール、エタノール、１−プロパノール、２−プロパノール、１−ブタノール、２−ブタノール等）、液状多価アルコール（１，３−ブチレングリコール、プロピレングリコール、グリセリン等）、ケトン類（アセトン、メチルエチルケトン等）、アセトニトリル、エステル類（酢酸エチル、酢酸ブチル等）、炭化水素類（ヘキサン、ヘプタン、流動パラフィン等）、エーテル類（エチルエーテル、テトラヒドロフラン、プロピルエーテル等）が挙げられる。好ましくは、水、低級１価アルコール及び液状多価アルコールが良く、特に好ましくは、水、エタノール、１，３−ブチレングリコール及びプロピレングリコールが良い。これらの溶媒は１種でも２種以上を混合して用いても良い。
【００１３】
また、メハジキ、カンゾウ、ジオウ、ロクジョウの抽出物は、抽出した溶液のまま用いても良く、必要に応じて、濃縮、希釈、濾過等の処理をして用いても良い。更には、抽出した溶液を濃縮乾固、噴霧乾燥、凍結乾燥等の処理を行い、乾燥物として用いても良い。
【００１４】
本発明の活性酸素消去酵素の活性促進剤は、上記抽出物をそのまま使用しても良く、メハジキ、カンゾウ、ジオウ、ロクジョウの抽出物の効果を損なわない範囲内で、通常の食品、化粧品、医薬部外品、医薬品に用いられる成分である油脂類、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、金属石鹸、pH調整剤、防腐剤、香料、保湿剤、粉体、紫外線吸収剤、増粘剤、色素、酸化防止剤、美白剤、キレート剤、賦形剤、安定剤、保存剤、結合剤、崩壊剤等の成分を配合することもできる。
【００１５】
本発明の活性酸素消去酵素の活性促進剤は食品、化粧品、医薬部外品及び医薬品のいずれにも用いることができ、その剤型としては、例えば、錠菓、飲料、化粧水、クリーム、乳液、ゲル剤、エアゾール剤、軟膏、パップ剤、ペースト剤、プラスター剤、エッセンス、パック、洗浄剤、浴用剤、ファンデーション、打粉、口紅、散剤、丸剤、錠剤、注射剤、坐剤、乳剤、カプセル剤、顆粒剤、液剤（チンキ剤、流エキス剤、酒精剤、懸濁剤、リモナーデ剤などを含む）等が挙げられる。
【００１６】
【実施例】
次に本発明を詳細に説明するため、本発明に用いる抽出物の製造例及び実験例を挙げるが、本発明はこれに限定されるものではない。
【００１７】
製造例１ メハジキの熱水抽出物
メハジキの地上部の全草２０ｇに４００ｍＬの精製水を加え、９５〜１００℃で２時間抽出した後、濾過し、その濾液を濃縮し、凍結乾燥してメハジキの熱水抽出物を２．２ｇ得た。
【００１８】
製造例２ カンゾウの熱水抽出物
カンゾウの根の乾燥物２０ｇに４００ｍＬの精製水を加え、９５〜１００℃で２時間抽出した後、濾過し、その濾液を濃縮し、凍結乾燥してカンゾウの熱水抽出物を１．０ｇ得た。
【００２０】
製造例４ ジオウの熱水抽出物
ジオウの根の乾燥物２０ｇに４００ｍＬの精製水を加え、９５〜１００℃で２時間抽出した後、濾過し、その濾液を濃縮し、凍結乾燥してジオウの熱水抽出物を１．２ｇ得た。
【００２１】
製造例５ ロクジョウの熱水抽出物
マンシュウジカの袋角の乾燥物２０ｇに４００ｍＬの精製水を加え、９５〜１００℃で２時間抽出した後、濾過し、その濾液を濃縮し、凍結乾燥してロクジョウの熱水抽出物を０．３ｇ得た。
【００２２】
製造例６ メハジキのエタノール抽出物
メハジキの全草の乾燥物１００ｇに９００ｍＬのエタノールを加え、常温で７日間抽出した後、濾過し、その濾液を濃縮乾固して、メハジキのエタノール抽出物を７．５ｇ得た。
【００２３】
製造例７ カンゾウのエタノール抽出物
カンゾウの全草の乾燥物１００ｇに９００ｍＬのエタノールを加え、常温で７日間抽出した後、濾過し、その濾液を濃縮乾固して、カンゾウのエタノール抽出物を６．０ｇ得た。
【００２５】
製造例９ ジオウのエタノール抽出物
ジオウの全草の乾燥物１００ｇに９００ｍＬのエタノールを加え、常温で７日間抽出した後、濾過し、その濾液を濃縮乾固して、ジオウのエタノール抽出物を６．２ｇ得た。
【００２６】
製造例１０ ロクジョウのエタノール抽出物
マンシュウアカジカの袋角の乾燥物１００ｇに９００ｍＬのエタノールを加え、常温で７日間抽出した後、濾過し、その濾液を濃縮乾固して、ロクジョウのエタノール抽出物を０．８ｇ得た。
【００２７】
製造例１１ メハジキの１，３−ブチレングリコール水溶液抽出物
メハジキの地上部全草の乾燥物１００ｇに１ｋｇの精製水及び１ｋｇの１，３−ブチレングリコールを加え、常温で７日間抽出した後、濾過し、メハジキの１，３−ブチレングリコール水溶液抽出物を１．９ｋｇ得た。
【００２８】
製造例１２ カンゾウの１，３−ブチレングリコール水溶液抽出物
カンゾウの根の乾燥物１００ｇに１ｋｇの精製水及び１ｋｇの１，３−ブチレングリコールを加え、常温で７日間抽出した後、濾過し、カンゾウの１，３−ブチレングリコール水溶液抽出物を１．９ ｋｇ得た。
【００３０】
製造例１４ ジオウの１，３−ブチレングリコール水溶液抽出物
ジオウの根の乾燥物１００ｇに１ｋｇの精製水及び１ｋｇの１，３−ブチレングリコールを加え、常温で７日間抽出した後、濾過し、ジオウの１，３−ブチレングリコール水溶液抽出物を１．９ｋｇ得た。
【００３１】
製造例１５ ロクジョウの１，３−ブチレングリコール水溶液抽出物
マンシュウジカの袋角の乾燥物１００ｇに１ｋｇの精製水及び１ｋｇの１，３−ブチレングリコールを加え、常温で７日間抽出した後、濾過し、ロクジョウの１，３−ブチレングリコール水溶液抽出物を１．９ｋｇ得た。
【００３２】
次に、本発明の効果を詳細に説明するため、実験例を挙げる。
【００３３】
実験例１ 表皮細胞内活性酸素消去酵素の活性促進作用
マウスケラチノサイト由来Pam212細胞1×105個を6cmディッシュに播種し、メハジキ、カンゾウ、ジオウ、ロクジョウの熱水抽出物を1μg/ml濃度に添加した10%
FCSを含むEagle’s MEMにて5% CO2、37℃条件下で4日間培養した。次に、細胞をPBS(-)にて2回洗浄した後、ラバーポリスマンにて剥離し、500μlのPBS(-)とともに超音波破砕したものを酵素試料液とした。酵素試料液中のSOD活性の測定はSODテストワコー（和光純薬工業株式会社製）を用いて行った。すなわち、酵素試料液100μl、発色試液450μlと酵素液（キサンチンオキシダーゼ）またはブランク液（キサンチンオキシダーゼの溶解に用いたリン酸緩衝液）450μlを混合し、37℃で20分間反応させた後、反応停止液1mlを添加し、560nmにおける吸光度を測定した。また、カタラーゼ活性の測定は過酸化水素の分解量を指標に行った。すなわち、10mMの過酸化水素を含む50mMリン酸緩衝液1mlに50μlの酵素試料液を添加し、240nmにおける1分間の吸光度変化を測定した。コントロールとしては試料無添加で培養した細胞を超音波破砕したものを用いた。なお、今回の実験で培地中に添加したメハジキ、カンゾウ、ジオウ、ロクジョウの熱水抽出物自体が酵素試料液のSODおよびカタラーゼ活性の測定結果に影響を及ぼさないことを確認している。
【００３４】
これらの試験結果は各試料を添加して培養した細胞のSODおよびカタラーゼ活性をコントロールを100とした場合の値で表１に示した。メハジキ、カンゾウ、ジオウ、ロクジョウの熱水抽出物はマウスケラチノサイト由来Pam212細胞におけるSODおよびカタラーゼ活性の促進作用を示した。特に、メハジキの熱水抽出物は SOD 活性の促進作用が高く、カンゾウ、ジオウ、ロクジョウの熱水抽出物はカタラーゼ活性の促進作用が高かった。
【００３５】
【表１】
製造例６〜１５についても同様の試験を行い、同様のＳＯＤおよびカタラーゼ活性の促進作用を示した。以上の実験例１の結果において、特にメハジキ及びジオウの抽出物が活性酸素消去酵素の活性促進作用が強かった。また、上記のいずれの抽出物も安定性に優れていた。
【００３７】
【発明の効果】
以上のことから、本発明のメハジキを含有する活性酸素消去酵素の活性促進剤、またはメハジキ、カンゾウ、ジオウ、ロクジョウの抽出物を一種又は二種以上含有するカタラーゼ活性促進剤は安定性に優れ、優れた活性酸素消去酵素の活性促進作用を示した。これらは、皮膚のみならず、活性酸素消去酵素が絡むあらゆる臓器についても同様である。[0001]
BACKGROUND OF THE INVENTION
TECHNICAL FIELD The present invention relates to an activity promoter for active oxygen scavenging enzyme, and in particular, active oxygen scavenging enzyme effective for preventing oxidation of living bodies including skin by promoting the activity of active oxygen scavenging enzyme which has been reduced by ultraviolet rays or aging. It relates to an activity promoter.
[0002]
[Prior art]
The skin is located in the outermost layer of the living body, is an organ in which active oxygen is easily generated due to the influence of ultraviolet rays and the like, and is constantly exposed to the oxygen stress. On the other hand, active oxygen scavenging enzymes exist in the skin cells, and protect the skin cells from injury of active oxygen unless active oxygen exceeding that capacity is generated. However, it is known that the activity of scavenging reactive oxygen scavenging enzymes in skin cells decreases with aging, and when the injury due to active oxygen surpasses its protective response, the skin is oxidized and the cellular function deteriorates and ages. It is thought to go. Accordingly, active oxygen scavengers and antioxidants have been studied as disclosed in JP-A-9-118630 and JP-A-9-208484 for the purpose of protecting against active oxygen injury, and active oxygen scavenging enzymes such as SOD and catalase, SOD Foods, cosmetics, quasi-drugs or pharmaceuticals containing active oxygen scavengers and antioxidants such as active substances have been developed.
[0003]
[Problems to be solved by the invention]
However, active oxygen scavenging enzymes such as SOD and catalase have problems in stability, and many active oxygen scavengers and antioxidants are not effective enough, so they are excellent in stability and highly effective in scavenging active oxygen. There is a need for enzyme activity promoters.
[0004]
[Means for Solving the Problems]
Under such circumstances, as a result of intensive investigations to obtain an active oxygen scavenging enzyme activity promoter excellent in protective action against active oxygen injury, the present inventors have found that extracts of Japanese pheasant, licorice, elephant and rouge are The inventors have found that the activity of the active oxygen scavenging enzyme inherent in the skin cells is stable and the present invention has been completed.
[0005]
That is, the present invention is an active oxygen scavenging enzyme activity promoter containing an extract of Japanese pheasant, licorice, dianthus, and rhododendron .
[0006]
The pheasant used in the present invention (scientific name: Leonurus sibiricus L.) is a herb belonging to the genus Lamiaceae, and grows naturally in sunny places in Yamano from Honshu, Shikoku, Kyushu, Korea and China to Southeast Asia. The whole plant of the territory of Mehajiki is the name of Yakumosou, and is used as a herbal medicine for postpartum hemostasis, menstrual irregularities, dizziness and abdominal pain.
[0007]
The licorice used in the present invention (scientific name: Glycyrrhiza uralensis) is a herb belonging to the genus Leguminosae and is distributed from Africa, Europe to Asia, Australia and western North America. Roots and running stems (strons) are used in Chinese medicine as licorice. You may use closely related licorice (scientific name: Glycyrrhiza glabra).
[0009]
Giant (scientific name: Rehmannia glutinosa Liboschitz) used in the present invention is a herb belonging to the genus Giantiaceae, and is distributed in North China and Korea. What dried the roots of Jiou is called ground yellow and is used as a herbal medicine.
[0010]
Locust used in the present invention is a deer family of Mandeka deer (scientific name: Cervus (Cervus) elephus L. var. Xanthhopygus Milne-Edwards) and Manshu deer (scientific name: Cervus (Sika) nippin t. It is a young horn (bag angle) that is not cornified yet or slightly keratinized.
[0011]
The extract of Japanese pheasant used in the present invention is extracted from a part or whole plant of a plant such as a leaf, stem, bark, flower, fruit, root, etc. Preferably, the one obtained by extracting from the whole plant of the above-ground part of a pheasant used as a crude drug is good. The extract of licorice is extracted from a part of the plant body such as leaves, stems, bark, flowers, fruits, roots or whole plants of licorice. Preferably, licorice roots and strons used as herbal medicines are extracted from dried ones. Diou extract is extracted from a part of the plant body such as leaves, stems, bark, flowers, fruits, roots, or the whole grass. Preferably, the dried roots of sage used as a crude drug are good. The extract of Rokujou is extracted from the horn of Manshu deer and Manchu deer. The preparation method is not particularly limited, and for example, it may be extracted by heating or may be extracted at room temperature. In addition, some of the above extracts are commercially available, and these can be used.
[0012]
Examples of the solvent to be extracted include water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohols (1,3-butylene glycol, Propylene glycol, glycerin, etc.), ketones (acetone, methyl ethyl ketone, etc.), acetonitrile, esters (ethyl acetate, butyl acetate, etc.), hydrocarbons (hexane, heptane, liquid paraffin, etc.), ethers (ethyl ether, tetrahydrofuran, Propyl ether, etc.). Water, lower monohydric alcohol and liquid polyhydric alcohol are preferable, and water, ethanol, 1,3-butylene glycol and propylene glycol are particularly preferable. These solvents may be used alone or in combination of two or more.
[0013]
In addition, the extract of Japanese pheasant, licorice, elephant, and rhododendron may be used as it is, or may be used after being subjected to treatments such as concentration, dilution, and filtration as necessary. Further, the extracted solution may be subjected to a treatment such as concentration to dryness, spray drying, freeze drying, etc., and used as a dried product.
[0014]
The active oxygen scavenging enzyme activity promoter of the present invention may use the above extract as it is, and within the range that does not impair the effects of the extract of pheasant, licorice, territory, and rhododendron , ordinary foods, cosmetics, pharmaceuticals Fats and oils, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, pH adjusters, antiseptics, perfumes, moisturizers, powders Components such as a body, an ultraviolet absorber, a thickener, a dye, an antioxidant, a whitening agent, a chelating agent, an excipient, a stabilizer, a preservative, a binder, and a disintegrant can also be blended.
[0015]
The active oxygen scavenging enzyme activity promoter of the present invention can be used in any of foods, cosmetics, quasi-drugs, and pharmaceuticals. Examples of the dosage form include tablets, beverages, lotions, creams, and emulsions. , Gel, aerosol, ointment, poultice, paste, plaster, essence, pack, detergent, bath preparation, foundation, dusting, lipstick, powder, pill, tablet, injection, suppository, emulsion, capsule Agents, granules, liquids (including tinctures, fluid extracts, spirits, suspensions, limonades, etc.).
[0016]
【Example】
Next, in order to describe the present invention in detail, examples of production and experimental examples of the extract used in the present invention are given, but the present invention is not limited thereto.
[0017]
Production Example 1 Hot water extract of Japanese pheasant 400 mL of purified water was added to 20 g of the whole plant of the terrestrial pheasant, extracted at 95-100 ° C. for 2 hours, filtered, the filtrate was concentrated, freeze-dried, Of hot water extract was obtained.
[0018]
Production Example 2 Hot water extract of licorice 400 mL of purified water was added to 20 g of dried licorice root, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, freeze-dried and dried with licorice. 1.0 g of hot water extract was obtained.
[0020]
Production Example 4 Geothermal hot water extract 400 g of purified water was added to 20 g of dried geous root, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate was concentrated, lyophilized and dried. 1.2 g of hot water extract was obtained.
[0021]
Production Example 5 Extract from hot water of Antarcticus 400 mL of purified water is added to 20 g of dried sachet of Manchu deer, extracted at 95-100 ° C. for 2 hours, filtered, and the filtrate is concentrated and freeze-dried. 0.3 g of hot water extract of Rokujou was obtained.
[0022]
Manufacture example 6: Extract of a Japanese reptile, added to 900 g of dried dry vegetative herb, and extracted at room temperature for 7 days, filtered, and concentrated to dryness. .5 g was obtained.
[0023]
Production Example 7 Licorice ethanol extract To 100 g of dried licorice whole plant, 900 mL of ethanol was added, extracted at room temperature for 7 days, filtered, the filtrate was concentrated to dryness, and licorice ethanol extract 6 0.0 g was obtained.
[0025]
Production Example 9 Diou ethanol extract 900 g of ethanol was added to 100 g of a dried product of Diou whole plant, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain an ethanol extract of Diou. 0.2 g was obtained.
[0026]
Production Example 10 Ethanol extract of Rokujou Add 900 mL of ethanol to 100 g of dried sardine of Manchua deer, extract for 7 days at room temperature, filter, concentrate the filtrate to dryness, 0.8g was obtained.
[0027]
Production Example 11 Extract from 1,3-butylene glycol aqueous solution of pheasant fish 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried terrestrial herb whole grass, extracted for 7 days at room temperature, and then filtered. As a result, 1.9 kg of a 1,3-butylene glycol aqueous solution extract of swordfish was obtained.
[0028]
Production Example 12 Extract of 1,3-butylene glycol aqueous solution of licorice 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried licorice root, extracted for 7 days at room temperature, filtered, and licorice 1.9 kg of 1,3-butylene glycol aqueous solution extract was obtained.
[0030]
Production Example 14 Diou 1,3-butylene glycol aqueous solution extract 1 kg of purified water and 1 kg of 1,3-butylene glycol were added to 100 g of dried diou root, and extracted at room temperature for 7 days. 1.9 kg of a 1,3-butylene glycol aqueous solution extract was obtained.
[0031]
Production Example 15 1,3-Butyleneglycol aqueous solution extract of Aspergillus added 1 kg of purified water and 1 kg of 1,3-butyleneglycol to 100 g of dried sachet of Manju deer, extracted for 7 days at room temperature, and then filtered. 1.9 kg of a 1,3-butylene glycol aqueous solution extract of Rokujou was obtained.
[0032]
Next, experimental examples will be given to explain the effects of the present invention in detail.
[0033]
Experimental Example 1 Promoting activity of reactive oxygen scavenging enzyme in epidermal cells 1 × 10 5 mouse keratinocyte-derived Pam212 cells were seeded in a 6cm dish, and hot water extracts of Japanese pheasant , licorice, elephant and rouge were added to a concentration of 1μg / ml 10%
The cells were cultured in Eagle's MEM containing FCS for 4 days under conditions of 5% CO 2 and 37 ° C. Next, the cells were washed twice with PBS (−), detached with a rubber policeman, and ultrasonically disrupted with 500 μl of PBS (−) as an enzyme sample solution. The SOD activity in the enzyme sample solution was measured using SOD Test Wako (manufactured by Wako Pure Chemical Industries, Ltd.). That is, 100 μl of enzyme sample solution, 450 μl of color reagent and 450 μl of enzyme solution (xanthine oxidase) or blank solution (phosphate buffer used to dissolve xanthine oxidase) were mixed and reacted at 37 ° C. for 20 minutes, and then the reaction was stopped. 1 ml of the solution was added, and the absorbance at 560 nm was measured. Catalase activity was measured using the amount of hydrogen peroxide decomposed as an index. That is, 50 μl of enzyme sample solution was added to 1 ml of 50 mM phosphate buffer containing 10 mM hydrogen peroxide, and the change in absorbance at 240 nm for 1 minute was measured. As a control, cells obtained by sonicating cells cultured without addition of a sample were used. In this experiment, it has been confirmed that the hot water extract of Japanese pheasant, licorice, terrestrial and cypress added to the medium itself does not affect the measurement results of SOD and catalase activity of the enzyme sample solution.
[0034]
These test results are shown in Table 1 with the SOD and catalase activity of the cells cultured with the addition of each sample as 100. The hot-water extracts of Japanese pheasant , licorice, Giant and Locust showed SOD and catalase activity in mouse keratinocyte-derived Pam212 cells. In particular, the hot-water extract of pheasant has a high SOD activity promoting effect, and the hot water extract of licorice, dianthus, and rhododendron has a high promoting activity of catalase activity.
[0035]
[Table 1]
The same tests were conducted on Production Examples 6 to 15 and showed similar SOD and catalase activity promoting effects. In the results of Experimental Example 1 described above, particularly, the extract of Japanese pheasant and dianthus had a strong activity promoting effect on the active oxygen scavenging enzyme. In addition, any of the above extracts was excellent in stability.
[0037]
【The invention's effect】
From the above, the active oxygen scavenging enzyme activity promoter containing the oysterfish of the present invention , or the catalase activity promoter containing one or more extracts of the oysterfish, licorice, dianthus, and rhododendron are excellent in stability, Excellent activity of scavenging enzyme for active oxygen scavenging enzyme was shown. The same applies not only to the skin but also to all organs that involve the active oxygen scavenging enzyme.
Claims (5)
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| JP5024978B2 (en) * | 2001-09-14 | 2012-09-12 | 丸善製薬株式会社 | Antioxidant |
| DE102005060880B4 (en) * | 2005-12-20 | 2012-04-05 | Universität Leipzig | Special extract for use as antiarrhythmic agent in cardiac arrhythmias and to improve coronary perfusion and its production |
| JP5464779B2 (en) * | 2006-04-26 | 2014-04-09 | 株式会社マンダム | Catalase activity inhibitor |
| KR100954386B1 (en) | 2007-09-17 | 2010-04-26 | 광동제약 주식회사 | The Manufacturing Process of extraction of the deer velvet and the deer velvet extract manufactured by the same method |
| JP2011020954A (en) * | 2009-07-15 | 2011-02-03 | Food Industry Research & Development Inst | Extract of eleutherococcus spp, method for preparing the same and use |
| CN102895334A (en) * | 2011-07-29 | 2013-01-30 | 苏州知微堂生物科技有限公司 | Preparation technology and production method for integrated new formulation of licorice and monkshood decoction |
| CN113768833A (en) * | 2021-08-16 | 2021-12-10 | 楚香(上海)生物科技有限公司 | Pure natural deacetylase activator and application thereof |
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| JP3195684B2 (en) * | 1993-03-01 | 2001-08-06 | 丸善製薬株式会社 | Skin cosmetics |
| JP3513872B2 (en) * | 1999-01-26 | 2004-03-31 | 株式会社コーセー | External preparation for skin |
| JP2000327550A (en) * | 1999-05-19 | 2000-11-28 | Kose Corp | Skin preparation for external use |
| JP2001181147A (en) * | 1999-12-27 | 2001-07-03 | Lion Corp | Cosmetic for hair |
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