JP6513469B2 - The external preparation for skin and the internal preparation containing the extract of the Chinese cabbage which is grown by irradiating the light which has a specific wavelength range. - Google Patents
The external preparation for skin and the internal preparation containing the extract of the Chinese cabbage which is grown by irradiating the light which has a specific wavelength range. Download PDFInfo
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- JP6513469B2 JP6513469B2 JP2015092736A JP2015092736A JP6513469B2 JP 6513469 B2 JP6513469 B2 JP 6513469B2 JP 2015092736 A JP2015092736 A JP 2015092736A JP 2015092736 A JP2015092736 A JP 2015092736A JP 6513469 B2 JP6513469 B2 JP 6513469B2
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Landscapes
- Cultivation Of Plants (AREA)
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- Medicines Containing Plant Substances (AREA)
Description
本発明は、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果、セラミド産生促進効果などに優れた新規な皮膚外用剤又は内用剤に関する。 TECHNICAL FIELD The present invention relates to a novel external skin or internal preparation excellent in antioxidative effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect, ceramide production promoting effect and the like.
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、皮膚細胞内の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。また、皮膚以外の臓器においても、その活性酸素消去能を越える活性酸素に曝されたとき、機能低下が起こり老化したり、ガンや心筋梗塞など様々な生活習慣病が発症すると考えられる。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、医薬部外品及び医薬品等が開発されている（特許文献１，２参照）。 The skin is located in the outermost layer of a living body, is an organ which is apt to generate active oxygen under the influence of ultraviolet light and the like, and is constantly exposed to the oxygen stress. On the other hand, there is an active oxygen scavenging enzyme in skin cells, and it protects skin cells from active oxygen damage unless active oxygen exceeding its capacity is generated. However, it is known that the activity of active oxygen scavenging enzymes in skin cells decreases with age, and when injury by active oxygen surpasses its protective response, the skin is oxidized, cell function is deteriorated and aging is caused. It is thought that it will continue. In addition, in organs other than the skin, when exposed to active oxygen exceeding its active oxygen scavenging ability, it is thought that functional decline occurs and aging or various lifestyle-related diseases such as cancer and myocardial infarction develop. Therefore, active oxygen scavenging agents and antioxidants are studied for the purpose of protection from injury by active oxygen, and reactive oxygen scavenging enzymes such as SOD and catalase, SOD-like active substances, etc. are blended with antioxidants. Food, cosmetics, quasi-drugs, medicines, etc. have been developed (see Patent Documents 1 and 2).
皮膚は、紫外線、乾燥、寒冷、熱、薬物等の様々な物理的及び化学的ストレスに日々曝されている。その結果、皮膚の機能低下が引き起こされ、様々な皮膚の老化現象が顕在化する。皮膚の老化現象の一つに、しわがある。しわには、表皮性のしわと、真皮性のしわの二種類が存在することが知られている。表皮性のしわは小じわと呼ばれ、皮膚の乾燥により、表皮角質層中の水分量が低下することによって一時的に生じるしわである。小じわの改善方法としては、保湿効果を有する化粧品の使用が一般的である。一方、真皮性のしわは、太陽光線に含まれる紫外線や加齢によって形成されるしわである。その形成メカニズムとしては、紫外線や加齢による真皮線維芽細胞におけるコラーゲン合成能の低下や、マトリックスメタロプロテアーゼ（ＭＭＰ）の増加によるコラーゲンの分解促進が挙げられる。 The skin is exposed daily to various physical and chemical stresses such as UV light, dryness, cold, heat, drugs and the like. As a result, a decrease in the function of the skin is caused, and various skin aging phenomena become apparent. One of the skin aging phenomena is wrinkles. Two types of wrinkles are known: epidermal wrinkles and dermal wrinkles. Epidermal wrinkles are called fine wrinkles, which are wrinkles temporarily caused by the reduction of the amount of water in the stratum corneum of the epidermis due to dryness of the skin. The use of cosmetics having a moisturizing effect is common as a method of improving fine lines. On the other hand, dermal wrinkles are wrinkles formed by ultraviolet rays contained in sunlight or by aging. The formation mechanism includes a decrease in the ability to synthesize collagen in dermal fibroblasts due to ultraviolet light and aging, and promotion of collagen degradation due to an increase in matrix metalloproteinase (MMP).
乾燥に起因する表皮性のしわと真皮性のしわでは、組織学的形態、発症メカニズム、治療方法が異なり、紫外線や加齢により生じる真皮性のしわは、保湿効果を有する化粧品の使用によっては改善できない。 Epidermal wrinkles caused by dryness and dermal wrinkles differ in histological form, development mechanism, and treatment method, and dermal wrinkles caused by ultraviolet light and aging are improved by the use of cosmetics having a moisturizing effect Can not.
これまでに、紫外線によって生じる真皮性のしわを改善することを目的として、加水分解アーモンドを有効成分とする皮膚のしわ形成防止・改善剤（特許文献３）、ジョチョウケイ、テンキシ及びキセンウの抽出物を有効成分とする紫外線照射に起因するしわの改善剤（特許文献４）が報告されている。 So far, for the purpose of improving the dermal wrinkles caused by ultraviolet light, an agent for preventing and treating wrinkles on the skin comprising hydrolysed almonds as an active ingredient (Patent Document 3), extracts of jojokei, tenxi and rhinoceros An agent for improving wrinkles caused by ultraviolet irradiation (Patent Document 4) has been reported.
また、真皮には線維芽細胞やコラーゲンが存在し、Ｉ型コラーゲンが全体の８０％を占める。Ｉ型コラーゲンのほかにはＩＩＩ、Ｖ、ＸＩＩ及びＸＩＶ型コラーゲンの存在が知られている。しわやたるみの原因の一つとして、Ｉ型コラーゲンの減少があげられる。従って、Ｉ型コラーゲンの生成を促進させることが、しわ・たるみの予防・改善に有効であると考えられる。また、Ｉ型コラーゲンの生成促進は皮膚の創傷治癒の改善にも有効である。 In addition, fibroblasts and collagen exist in the dermis, and type I collagen accounts for 80% of the whole. Besides type I collagen, the presence of type III, V, XII and type XIV collagens is known. One of the causes of wrinkles and slackness is the reduction of type I collagen. Therefore, promoting the production of type I collagen is considered to be effective for the prevention and improvement of wrinkles and sags. In addition, promoting the production of type I collagen is also effective in improving wound healing of the skin.
また、線維芽細胞はコラーゲンなどのタンパク質を産生して真皮結合組織を形成し、皮膚のハリを保っている。この結合組織が収縮力を失い、さらに弾力性を失う結果として皮膚のシワやタルミが発生すると考えられている。 In addition, fibroblasts produce proteins such as collagen to form dermal connective tissue and maintain the firmness of the skin. It is believed that this connective tissue loses its contractility and loses its elasticity resulting in wrinkles and tears on the skin.
コラーゲンは、哺乳動物組織の約１/３を占める主要な構造タンパク質であり、軟骨、骨、腱、及び皮膚を含む多くのマトリックス組織の必須な成分である。マトリックスメタロプロテアーゼ（ＭＭＰ）に属するコラゲナーゼ（ＭＭＰ−１）により１箇所を切断されると、通常の組織内では安定なコラーゲン分子は、変性して一本鎖のゼラチンとなり、他の様々なプロテアーゼにより分解されるようになる。その結果、マトリックス組織の構造の完全性が失われてしまう。 Collagen is a major structural protein that accounts for about one third of mammalian tissue and is an essential component of many matrix tissues, including cartilage, bone, tendons, and skin. Once cleaved at one site by collagenase (MMP-1) belonging to matrix metalloproteinases (MMPs), stable collagen molecules in normal tissues are denatured into single-stranded gelatin and by various other proteases It becomes disassembled. As a result, the structural integrity of the matrix tissue is lost.
一般に、シミ、ソバカス、日焼け等に見られる皮膚の色素沈着は、ホルモンの異常や紫外線の刺激により、皮膚内に存在するメラノサイトがメラニン色素を過剰に生成し、これが皮膚内に沈着することが原因と考えられている。このような色素沈着を防ぐ方法の一つに、メラニンの過剰な生成を抑制する方法が知られている。従来、色素沈着の治療には、内用や外用などにおいて、アスコルビン酸（ビタミンＣ）等が用いられてきた。 In general, pigmentation of the skin observed in spots, buckwheat, sunburn, etc. is caused by the fact that melanocytes present in the skin excessively generate melanin pigment due to hormonal abnormalities or stimulation of ultraviolet light, and this is deposited in the skin. It is believed that. One of the methods to prevent such pigmentation is known to suppress excessive production of melanin. Conventionally, ascorbic acid (vitamin C) and the like have been used for internal and external use for the treatment of pigmentation.
加齢とともに表皮細胞の増殖・分裂能は低下し、表皮層自体は薄くなる（非特許文献１参照）。生体因子であるＥｐｉｄｅｒｍａｌ Ｇｒｏｗｔｈ Ｆａｃｔｏｒ（ＥＧＦ／上皮細胞成長因子）や女性ホルモン（エストロゲン）は皮膚の表皮細胞増殖に働きかけるが、加齢と共にその分泌は低下する。このような加齢による表皮細胞代謝機能の低下は、皮膚のターンオーバー速度を遅らせ、肌荒れや皮膚の老化の原因となる。また、角層表面から剥がれ落ちる角層細胞が滞留することで、表皮内メラニンの排泄がスムーズに行われなくなり、色素沈着や肌のくすみの原因となる。さらに表皮の創傷治癒が遅くなることなども知られている。これらの現象の進行を防止あるいは改善するために、表皮細胞の増殖を促進させる成分の探索や、多くの皮膚外用剤の提案がなされてきた。 The ability of epidermal cells to proliferate and divide decreases with age, and the epidermal layer itself becomes thin (see Non-patent Document 1). Biological factors, such as the Growth Growth Factor (EGF / epithelial growth factor) and the female hormone (estrogen), work on epidermal cell proliferation in the skin, but their secretion decreases with age. Such a decrease in epidermal cell metabolic function due to aging slows the turnover speed of the skin and causes rough skin and aging of the skin. In addition, since stratum corneum cells falling off from the stratum corneum surface stay, the excretion of melanin in the epidermis can not be smoothly performed, which causes pigmentation and skin dullness. Furthermore, it is also known that wound healing of the epidermis is delayed. In order to prevent or ameliorate the progression of these phenomena, a search has been made for a component that promotes the proliferation of epidermal cells, and many external skin preparations have been proposed.
セラミドは、スフィンゴ脂質の一種であり、スフィンゴシンと脂肪酸がアミド結合した化合物群の総称である。セラミドは、細胞膜において高濃度で存在することが知られている。又、セラミドは、角質細胞間脂質の構成成分の一種であり、角化の過程において細胞外に分泌され、皮膚のバリア機能や水分保持能に重要な役割を果たしている。更に、セラミドは、シグナル伝達物質として、細胞の増殖、分化及びアポトーシス等を制御することが知られている。これらのことから、セラミドの産生を促進する物質には、細胞の増殖抑制、分化誘導及びアポトーシスの誘導効果等が期待でき、これらの異常に起因する疾患に対する治療効果が期待できると考えられる。
セラミドとの関連性が高い皮膚疾患として、例えば、扁平上皮癌、乾癬及びアトピー性皮膚炎等が挙げられる。これらのことから、皮膚におけるセラミドの産生を促進することによって、扁平上皮癌、乾癬及びアトピー性皮膚炎等の皮膚疾患を治療及び予防することが可能である。
Ceramide is a type of sphingolipid and is a generic name of a group of compounds in which sphingosine and a fatty acid form an amide bond. Ceramide is known to be present at high concentrations in cell membranes. In addition, ceramide is one of the components of inter-keratinocyte lipid, and is secreted extracellularly in the process of keratinization, and plays an important role in the barrier function and water retention ability of the skin. Furthermore, ceramide is known as a signal transduction substance to control cell proliferation, differentiation, apoptosis and the like. From these facts, it is considered that substances that promote the production of ceramide can be expected to suppress cell growth, induce differentiation and induce apoptosis, etc., and to expect therapeutic effects for diseases caused by these abnormalities.
Skin diseases highly associated with ceramide include, for example, squamous cell carcinoma, psoriasis and atopic dermatitis. From these facts, it is possible to treat and prevent skin diseases such as squamous cell carcinoma, psoriasis and atopic dermatitis by promoting the production of ceramide in the skin.
ヒト皮膚には、７系統のセラミドが存在することが確認されており、全種類のセラミドが角質層に存在する比率で補うことが理想的である。従来、この様な肌荒れ、乾燥肌の予防改善に有効な化粧料として、セラミドが種々の皮膚外用剤に配合されているが、ヒトの皮膚に存在する全種類のセラミドを適正な比率で補充することは技術的に困難であった。したがって、生体内におけるセラミド産生を促進する事が重要であると考えられる。セラミド産生促進剤としては、特許文献５や特許文献６が報告されている。 Seven types of ceramides have been confirmed to be present in human skin, and it is ideal to compensate for all types of ceramides in the ratio in the stratum corneum. Conventionally, ceramide is blended in various external skin preparations as a cosmetic effective for preventing and improving such rough skin and dry skin, but all kinds of ceramide present in human skin are replenished at an appropriate ratio. It was technically difficult. Therefore, it is considered important to promote ceramide production in vivo. Patent documents 5 and 6 have been reported as ceramide production promoters.
クウシンサイの公知文献としては、タンパク質糖化抑制能（特許文献７）などが知られていた。 As a known reference of kuusinsai, the protein saccharification suppression ability (patent document 7) etc. were known.
一方で、植物の栽培方法によって植物の薬効を高める方法として、植物体内のビタミンやポリフェノール、ルチンなどの機能性物質を特徴的に増加させる方法は、すでに特許文献で報告されている。特許文献８には、大豆もやしに近紫外〜青色領域波長の光を照射することにより、含有ビタミンＡ、ビタミンＥを増量させる方法が開示されており、特許文献９には、小松菜に対して、人工紫外線照射を１日５分間行うことで、機能性物質であるα−トコフェロールやビタミンＣを増加させる栽培方法が開示され、特許文献１０には、人工光源の青色光、赤色光及び遠赤色光の強度を調整することにより、小松菜、レタスのビタミンＣやビタミンＡを増加させる方法が開示されている。 On the other hand, methods for characteristically increasing functional substances such as vitamins, polyphenols and rutin in the plant body have already been reported in patent documents as methods for enhancing the medicinal effects of plants by cultivation methods of plants. Patent Document 8 discloses a method of increasing contained vitamin A and vitamin E by irradiating soy bean sprouts with light of near ultraviolet to blue wavelength, and Patent Document 9 discloses a method for treating komatsuna vegetables. A cultivation method for increasing the functional substance α-tocopherol and vitamin C by performing artificial ultraviolet irradiation for 5 minutes a day is disclosed. Patent Document 10 discloses blue light, red light and far red light of artificial light source There is disclosed a method of increasing vitamin C and vitamin A of komatsuna and lettuce by adjusting the strength of
本発明は、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果、セラミド産生促進効果などに優れた新規な皮膚外用剤又は内用剤を提供することを課題とする。 The present invention provides a novel external skin or internal preparation excellent in antioxidative effect, collagen formation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect, ceramide production promoting effect, etc. As an issue.
本発明者らは、この問題点を解決すべく、鋭意研究を重ねた結果、特定の波長域を有する２種の光を同時に照射して栽培したクウシンサイの抽出物に、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果、セラミド産生促進効果が優れていることを発見し、本発明を完成するに至った。 As a result of intensive studies to solve this problem, the present inventors have an antioxidant effect and collagen formation on the extract of Chinese cabbage grown by simultaneously irradiating two types of light having a specific wavelength range. The inventors have found that the promoting effect, the matrix metalloproteinase (MMP) inhibiting effect, the whitening effect, the cell proliferation effect, and the ceramide production promoting effect are excellent, and the present invention has been completed.
すなわち、本発明は、以下の（１）〜（７）からなる。 That is, the present invention consists of the following (1) to (7).
（１）波長域５７０〜７３０ｎｍと４００〜５１５ｎｍとの光合成光量子束密度（ＰＰＦＤ）比が８:１〜１：１の光を照射して栽培したクウシンサイの、水、低級アルコール及び液状多価アルコールから選ばれる一種又は二種以上の溶媒による抽出物を含有することを特徴とする皮膚外用剤。
（２）波長域５７０〜７３０ｎｍと４００〜５１５ｎｍとの光合成光量子束密度（ＰＰＦＤ）比が、４:１〜２:１であることを特徴とする請求項１記載の皮膚外用剤。
（３）波長域５７０〜７３０ｎｍと４００〜５１５ｎｍとの光合成光量子束密度（ＰＰＦＤ）比が８:１〜１：１の光を照射して栽培することによって、蛍光灯又は太陽光で栽培したクウシンサイと比較して、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果及びセラミド産生促進効果から選ばれる一種又は二種以上の効果を高めることを特徴とするクウシンサイ。
（４）波長域５７０〜７３０ｎｍと４００〜５１５ｎｍとの光合成光量子束密度（ＰＰＦＤ）比が８:１〜１：１の光を照射して栽培することによって、蛍光灯又は太陽光で栽培したクウシンサイと比較して、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果及びセラミド産生促進効果から選ばれる一種又は二種以上の効果を高めたクウシンサイ又は、水、低級アルコール及び液状多価アルコールから選ばれる一種又は二種以上の溶媒によって抽出される抽出物を含有することを特徴とする医薬品。
（５）波長域５７０〜７３０ｎｍと４００〜５１５ｎｍとの光合成光量子束密度（ＰＰＦＤ）比が８:１〜１：１の光を照射して栽培することによって、蛍光灯又は太陽光で栽培したクウシンサイと比較して、抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果及びセラミド産生促進効果から選ばれる一種又は二種以上の効果を高めたクウシンサイ又は、水、低級アルコール及び液状多価アルコールから選ばれる一種又は二種以上の溶媒によって抽出される抽出物を含有することを特徴とする食品。
（６）波長域５７０〜７３０ｎｍ及び／又は４００〜５１５ｎｍの光を照射して栽培したクウシンサイの、水、低級アルコール及び液状多価アルコールから選ばれる一種又は二種以上の溶媒によって抽出される抽出物を含有することを特徴とする皮膚外用剤。
（７）波長域５７０〜７３０ｎｍ及び／又は４００〜５１５ｎｍの光を照射して栽培したクウシンサイ、又は、そのクウシンサイの水、低級アルコール及び液状多価アルコールから選ばれる一種又は二種以上の溶媒によって抽出される抽出物。
(1) Water, lower alcohol and liquid polyhydric alcohol of a Chinese cabbage grown by irradiating with light having a photosynthetic photon flux density (PPFD) ratio of 8: 1 to 1: 1 between wavelength regions 570 to 730 nm and 400 to 515 nm An external preparation for skin comprising an extract with one or more solvents selected from the group consisting of
(2) The external preparation for skin according to claim 1, wherein the ratio of photosynthetic photon flux density (PPFD) of the wavelength range of 570 to 730 nm and 400 to 515 nm is 4: 1 to 2: 1.
(3) A light source grown with a fluorescent lamp or sunlight by irradiating and cultivating a light having a photosynthetic photon flux density (PPFD) ratio of 8: 1 to 1: 1 between wavelength regions 570 to 730 nm and 400 to 515 nm. And one or more effects selected from antioxidative effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect and ceramide production promoting effect, as compared with Cucumber rhinoceros.
(4) A cultivar grown with a fluorescent lamp or sunlight by irradiating and cultivating light having a photosynthetic photon flux density (PPFD) ratio of 8: 1 to 1: 1 between wavelength regions 570 to 730 nm and 400 to 515 nm. Compared with the above, it has an effect of one or more selected from the antioxidant effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect and ceramide production promoting effect, or A pharmaceutical comprising the extract extracted by one or more solvents selected from water, lower alcohols and liquid polyhydric alcohols.
(5) A cultivar cultivated with a fluorescent lamp or sunlight by irradiating and cultivating light having a photosynthetic photon flux density (PPFD) ratio of 8: 1 to 1: 1 between wavelength regions 570 to 730 nm and 400 to 515 nm. Compared with the above, it has an effect of one or more selected from the antioxidant effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect and ceramide production promoting effect, or What is claimed is: 1. A food comprising an extract extracted by one or more solvents selected from water, lower alcohols and liquid polyhydric alcohols.
(6) An extract of a Chinese cabbage grown by irradiation with light in a wavelength range of 570 to 730 nm and / or 400 to 515 nm, which is extracted with one or more solvents selected from water, lower alcohol and liquid polyhydric alcohol A skin external preparation characterized by containing.
(7) Extracting with the seeds of Kushinsai grown by irradiation with light in the wavelength range of 570 to 730 nm and / or 400 to 515 nm, or one or more solvents selected from water, lower alcohol and liquid polyhydric alcohol Extract.
本発明のクウシンサイ又はその抽出物は、優れた抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果、セラミド産生促進効果を有しており、医薬品、医薬部外品、化粧品、食品の分野において貢献できるものである。 The kushishinsai of the present invention or its extract has excellent antioxidative effect, collagen formation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation effect, ceramide production promoting effect It can contribute in the field of accessories, cosmetics and food.
以下に、本発明について詳細に述べる。 Hereinafter, the present invention will be described in detail.
本発明に用いるクウシンサイの抽出物とは、ヒルガオ科サツマイモ属のクウシンサイ、別名ヨウサイ（学名：Ｉｐｏｍｏｅａ ａｑｕａｔｉｃａ）の花、実、種子、茎、葉、根等の植物体の一部又は全草から抽出したものである。その抽出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、本発明においては、抽出物の代わりに、植物体のまま使用することもでき、生のままでも、乾燥して用いることもでき、目的によって使い分けることができる。さらには、抽出物と植物体を併用することもできる。 The extract of Cupressus sinensis used in the present invention is extracted from a part or whole of a plant such as flowers, fruits, seeds, stems, leaves, roots, etc. of Cupressaceae that is a genus of the sweet potato genus Sweet potato genus, also known as Ipomoea aquatica It is The extraction method is not particularly limited, and for example, it may be extracted by heating or may be extracted at normal temperature. In the present invention, instead of the extract, the plant body can be used as it is, or it can be used as it is or dried and can be used properly depending on the purpose. Furthermore, the extract and a plant can be used in combination.
栽培方法としては、土を用いた栽培や水耕栽培で行うことができる。水耕栽培で行う場合には、種子を播種後、出根した状態で、水耕栽培に供することができる。栽培は、温度、光、二酸化炭素濃度が制御された施設で栽培することが好ましい。栽培温度は、１５〜３０℃、好ましくは２０〜２５℃である。栽培期間は、照射する条件によって異なるが、概ね１０〜３０日で収穫できる。これ以上栽培することも可能である。 As a cultivation method, it can carry out by cultivation and hydroponic cultivation which used soil. When carrying out by hydroponic cultivation, after sowing a seed, it can use for hydroponic cultivation in the state which rooted. Cultivation is preferably conducted at a facility where the temperature, light and carbon dioxide concentration are controlled. The cultivation temperature is 15 to 30 ° C, preferably 20 to 25 ° C. The cultivation period varies depending on the irradiation conditions, but can be harvested in approximately 10 to 30 days. It is also possible to cultivate more than this.
光源は、植物の栽培施設で用いる光源などを使用することができ、発光ダイオード（ＬＥＤ）、レーザーダイオードなどの光半導体素子があげられるが、特定の範囲の波長域が選択的に照射できる光源であればＬＥＤに限らない。 The light source may be a light source used in a plant cultivation facility, and may be an optical semiconductor element such as a light emitting diode (LED) or a laser diode, but a light source capable of selectively irradiating a wavelength range of a specific range. If it is, it is not restricted to LED.
クウシンサイの栽培において、照射する波長としては、波長域４００〜５１５ｎｍの青色光、５７０〜７３０ｎｍの赤色光であることが好ましく、波長域４３０〜４６０ｎｍ、６３０〜６８０ｎｍの光がさらに好ましい。これらの光は、同時に照射することが最も好ましい。このときの波長は、照射スペクトルの極大波長（ピーク波長）のことをいう。このような波長のピークを有する光源であれば、独自に作成したものや市販のものを使用することもできる。また、上記波長を選択的に照射できるように、光学フィルタを用いても良い。上記の２種の範囲の光に加え、太陽光や蛍光灯などの光源を使用することもできる。 In the cultivation of a Chinese cabbage, as a wavelength to irradiate, it is preferable that they are blue light of a wavelength range 400-515 nm, and red light of 570-730 nm, and light of a wavelength range 430-460 nm, 630-680 nm is still more preferable. It is most preferable that these lights be irradiated simultaneously. The wavelength at this time refers to the maximum wavelength (peak wavelength) of the irradiation spectrum. As long as it is a light source having such a peak of wavelength, it is also possible to use a uniquely created or commercially available one. In addition, an optical filter may be used so that the above wavelength can be selectively irradiated. In addition to the two ranges of light described above, light sources such as sunlight or fluorescent lamps can also be used.
照射する光量としては、光合成有効光量子束密度（ＰＰＦＤ）として表される。発光体を２種組み合わせて照射する場合には、その合計の光量を意味する。その光量は、発芽後は１０〜３００μｍｏｌ・ｍ−２ｓ−１が好ましく、５０〜２００μｍｏｌ・ｍ−２ｓ−１がさらに好ましい。この範囲外の光強度の場合は、生育障害、生育不良になる場合がある。照射は、クウシンサイの上部１０〜５０ｃｍの位置から照射することが好ましい。照射時間は、植物の特性や目的に応じて適宜変更できるが、６時間以上が好ましく、１２〜２４時間がより好ましい。 The amount of light to be irradiated is expressed as a photosynthetically available photon flux density (PPFD). In the case where two types of light emitters are combined and irradiated, the total amount of light is meant. Its quantity of light after germination is preferably 10~300μmol · m -2 s -1, more preferably 50~200μmol · m -2 s -1. If the light intensity is out of this range, it may cause growth failure and growth failure. The irradiation is preferably performed from the position of 10 to 50 cm on the upper part of the Chinese cabbage. Although irradiation time can be suitably changed according to the characteristic and purpose of a plant, 6 hours or more are preferable, and 12 to 24 hours are more preferable.
赤色と青色の光量比においては、それぞれのＰＰＦＤの比を意味しており、収量や有効性など目的に応じて選択が可能である。 The light amount ratio of red to blue means the ratio of each PPFD, and can be selected according to the purpose such as yield and effectiveness.
中でも、植物体の収量を高めるには、赤色と青色の光量比が８：１〜２：１に高い収量が得られた。 Above all, in order to increase the yield of plants, a high yield was obtained when the light amount ratio of red to blue was 8: 1 to 2: 1.
活性酸素消去作用（フリーラジカル捕捉除去作用）においては、赤色と青色の光量比が４：１〜０：１が効果の面で好ましく、３：１〜０：１がより好ましい。その中でも特に、赤色と青色の光量比が３：１が最も好ましい。 In the active oxygen scavenging action (free radical scavenging action), the light amount ratio of red to blue is preferably 4: 1 to 0: 1 in view of the effect, and more preferably 3: 1 to 0: 1. Among them, the light amount ratio of red to blue is particularly preferably 3: 1.
Ｉ型コラーゲン（ＣＯＬ１Ａ）発現促進作用においては、赤色と青色の光量比が８：１〜１：１が効果の面で好ましい。その中でも特に、赤色と青色の光量比が４：１〜２：１が最も好ましい。 In the type I collagen (COL1A) expression promoting action, the light amount ratio of red to blue is preferably 8: 1 to 1: 1 in view of the effect. Among them, the light amount ratio of red to blue is particularly preferably 4: 1 to 2: 1.
ＭＭＰ−１ ｍＲＮＡ発現抑制作用においては、赤色と青色の光量比が８：１〜２：１が効果の面で好ましい。その中でも特に、赤色と青色の光量比が４：１が最も好ましい。 In the MMP-1 mRNA expression suppression action, the light amount ratio of red to blue is preferably 8: 1 to 2: 1 in view of the effect. Among them, the light amount ratio of red to blue is particularly preferably 4: 1.
メラニン生成抑制作用においては、赤色と青色の光量比が３：１〜１：１が効果の面で好ましい。その中でも特に、赤色と青色の光量比が２：１が最も好ましい。 In the melanin production inhibitory action, the light amount ratio of red to blue is preferably 3: 1 to 1: 1 in view of the effect. Among them, the light amount ratio of red to blue is particularly preferably 2: 1.
細胞増殖促進作用においては、赤色と青色の光量比が３：１〜１：１が効果の面で好ましい。その中でも特に、赤色と青色の光量比が３：１〜２：１が最も好ましい。 In the cell growth promoting action, the light amount ratio of red to blue is preferably 3: 1 to 1: 1 in view of the effect. Among them, the light amount ratio of red to blue is particularly preferably 3: 1 to 2: 1.
セラミド生成促進作用においては、赤色と青色の光量比が８：１〜１：１が効果の面で好ましく、４：１〜２：１がより好ましい。その中でも特に、赤色と青色の光量比が４：１〜３：１が最も好ましい。 In the ceramide formation promoting action, the light amount ratio of red to blue is preferably 8: 1 to 1: 1 in view of the effect, and more preferably 4: 1 to 2: 1. Among them, the light amount ratio of red to blue is particularly preferably 4: 1 to 3: 1.
以上のことを総じていえば、赤色と青色の光量比が８：１〜１：１が好ましく、４：１〜２：１が最も好ましい。 Generally speaking, the light amount ratio of red to blue is preferably 8: 1 to 1: 1, and most preferably 4: 1 to 2: 1.
抽出溶媒としては、例えば、水、低級アルコール（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）、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、低級アルコール及び液状多価アルコール等の極性溶媒が良く、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。これらの溶媒は一種でも二種以上を混合して用いても良い。 As an extraction solvent, for example, water, lower alcohol (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol), liquid polyhydric alcohol (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.) It can be mentioned. Preferably, polar solvents such as water, lower alcohols and liquid polyhydric alcohols are preferred, and water, ethanol, 1,3-butylene glycol and propylene glycol are particularly preferred. These solvents may be used alone or in combination of two or more.
上記抽出物は、抽出した溶液のまま用いても良く、必要に応じて、濃縮、希釈及び濾過処理、活性炭等による脱色、脱臭処理等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。また、サラダなど、生で食することもできる。 The extract may be used as it is as the extracted solution, and if necessary, it may be used after concentration, dilution and filtration, decolorization with activated carbon and the like, deodorization and the like. Furthermore, the extracted solution may be subjected to treatments such as concentration to dryness, spray drying, lyophilization and the like, and used as a dried product. You can also eat fresh, such as salads.
本発明の外用剤又は内用剤には、食品も含むものとし、これには、上記植物体及び／又は抽出物をそのまま使用しても良く、これらの効果を損なわない範囲内で、化粧品、医薬部外品、医薬品又は食品等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、皮膜剤、甘味料、酸味料等の成分を配合することもできる。 The external preparation or internal preparation of the present invention includes food, and the above-mentioned plant body and / or extract may be used as it is, as long as the effects thereof are not impaired. Oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants, metal soaps, metal soaps, pH adjusters, preservatives, perfumes, moisturizers which are components used in non-prescription products, pharmaceuticals and foods Ingredients such as agents, powders, UV absorbers, thickeners, dyes, antioxidants, skin lightening agents, chelating agents, excipients, coatings, sweeteners, acidulants and the like can also be blended.
本発明は、医薬品、医薬部外品、化粧品、食品に用いることができ、その剤型としては、例えば、化粧水、クリーム、マッサージクリーム、乳液、ゲル剤、エアゾール剤、パック、洗浄剤、浴用剤、ファンデーション、打粉、口紅、軟膏、パップ剤、ペースト剤、プラスター剤、エッセンス、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、リモナーデ剤等を含む）、錠菓、飲料、ティーバッグ、スパイス等が挙げられる。 The present invention can be used for medicines, quasi-drugs, cosmetics, foods, and the dosage forms thereof include, for example, lotions, creams, massage creams, emulsions, gels, aerosols, packs, cleaning agents, and baths. Agent, foundation, dusting, lipstick, ointment, patch, paste, plaster, essence, powder, pill, tablet, injection, suppository, emulsion, capsule, granule, liquid (tincture, flow extract) Alcohol, suspending agents, limonade agents and the like), tablets, beverages, tea bags, spices and the like.
本発明に用いる上記抽出物の配合量は、外用の場合、全量に対し、固形物に換算して０．０００１重量％以上が好ましく、０．００１〜１０重量％がより好ましい。さらに、０．０１〜５重量％が最も好ましい。０．０００１重量％未満では十分な効果は望みにくい。１０重量％を越えて配合した場合、効果の増強は認められにくく不経済である。一方、内用の場合、投与量は年齢、体重、症状、治療効果、投与方法、処理時間等により異なるが、通常、成人１人当たりの１日の量としては、５ｍｇ以上が好ましく、１０ｍｇ〜５ｇがより好ましい。さらに、１００ｍｇ〜１ｇが最も好ましい。 The amount of the extract used in the present invention is preferably 0.0001% by weight or more, more preferably 0.001 to 10% by weight, in terms of solid matter, based on the total amount in the case of external use. Furthermore, 0.01 to 5% by weight is most preferable. If it is less than 0.0001% by weight, it is difficult to obtain a sufficient effect. When it is compounded in excess of 10% by weight, the effect is hardly enhanced and it is uneconomical. On the other hand, in the case of internal use, the dosage varies depending on the age, body weight, symptoms, treatment effect, administration method, treatment time etc. Usually, 5 mg or more is preferable as a daily dose per adult, and 10 mg to 5 g Is more preferred. Furthermore, 100 mg to 1 g is most preferable.
次に本発明を詳細に説明するため、実施例として本発明に用いるクウシンサイの抽出物の製造例、実験例及び処方例を挙げるが、本発明はこれに限定されるものではない。製造例に示す％とは重量％を、実施例に示す配合量の部とは重量部を示す。 Next, in order to explain the present invention in detail, although production examples, experimental examples and formulation examples of an extract of Chinese cabbage used in the present invention are given as examples, the present invention is not limited thereto. % Shown in a manufacture example shows weight%, and the part of the compounding quantity shown to an Example shows a weight part.
（１）実験材料および生育条件
水分を含んだメッシュにクウシンサイの種子を播種し、温度２２〜２４℃・暗所で発芽させ、これをスポンジに包み、２２〜２４℃で２４時間白色蛍光灯下で栽培し、育苗した。その後、水耕栽培装置を用いて、室温２１〜２３℃で２４時間、植物の真上３０ｃｍの位置から、青色ＬＥＤ（ピーク波長４５０ｎｍ）及び赤色ＬＥＤ（ピーク波長６６０ｎｍ）を同時に照射し、赤色と青色ＬＥＤの合計光合成有効光量子束密度１００μｍｏｌ・ｍ−２ｓ−１となるように、赤色と青色の光量比を１：０〜０：１にして、栽培を行った。また、比較例１として光合成有効光量子束密度１００μｍｏｌ・ｍ−２ｓ−１となるように白色蛍光灯下で栽培を行い、比較例２として太陽光下で栽培を行った。なお、栽培中は光量比を変えなかった。４週間栽培した後、収穫し、生クウシンサイを得た。これを約６０℃で温風乾燥させることで、クウシンサイの乾燥物を得た（表１）。
(1) Experimental materials and growth conditions Seeds of Chinese cabbage are sown on a moist mesh, germinated in the dark at a temperature of 22-24 ° C, wrapped in a sponge, and under white fluorescent light at 22-24 ° C for 24 hours Grown and raised. Then, using a hydroponic cultivation apparatus, simultaneously illuminate a blue LED (peak wavelength 450 nm) and a red LED (peak wavelength 660 nm) from a position 30 cm directly above the plant for 24 hours at room temperature 21-23 ° C. Cultivation was carried out with a light amount ratio of red to blue of 1: 0 to 0: 1 so as to obtain a total photosynthetic effective photon flux density of 100 μmol · m −2 s −1 of the blue LED. In addition, as Comparative Example 1, cultivation was performed under a white fluorescent lamp so as to obtain a photosynthetically effective photon flux density of 100 μmol · m −2 s −1, and as Comparative Example 2, cultivation was performed under sunlight. The light intensity ratio was not changed during cultivation. After four weeks of cultivation, it was harvested to obtain fresh cucumber. The product was dried with warm air at about 60 ° C. to obtain a dried product of kushishin (Table 1).
（２）抽出
製造例１Ａ 熱水抽出物
乾燥物１０ｇに精製水２００ｍＬ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、凍結乾燥して熱水抽出物を得た（表２）。
(2) Extraction
Production Example 1A Hot water extract Add 200 mL of purified water to 10 g of the dried product, extract at 95 to 100 ° C. for 2 hours, filter, concentrate the filtrate, lyophilize and extract the hot water extract Obtained (Table 2).
製造例１Ｂ ５０％エタノール抽出物
乾燥物１０ｇに５０％エタノール２００ｍＬ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、５０％エタノール抽出物を得た（表２）。
Production Example 1 B 50% ethanol extract 200 g of 50% ethanol was added to 10 g of the dried product, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain 50% ethanol extract. (Table 2).
製造例１Ｃ エタノール抽出物
乾燥物１０ｇにエタノール２００ｍＬ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、エタノール抽出物を得た（表２）。
Production Example 1 C Ethanol extract 200 mL of ethanol was added to 10 g of the dried product, extracted at room temperature for 7 days, filtered, and the filtrate was concentrated to dryness to obtain an ethanol extract (Table 2).
上記と同様に、赤色と青色ＬＥＤの合計光合成有効光量子束密度１００μｍｏｌ・ｍ−２ｓ−１となるように、赤色と青色の光量比を変化させて栽培したクウシンサイまたは比較例1として光合成有効光量子束密度１００μｍｏｌ・ｍ−２ｓ−１となるように白色蛍光灯下で上記の生育条件と同様に栽培したクウシンサイ、比較例２として太陽光で栽培したクウシンサイを用い、上記の製造例１Ａ〜１Ｃと同様に抽出し、製造例２Ａ〜７Ｃ、比較製造例１Ａ〜１Ｃ、比較製造例２Ａ〜２Ｃとした（表２）。 In the same manner as described above, the Chinese cabbage grown by changing the light amount ratio of red and blue so that the total photosynthetic effective photon flux density of red and blue LEDs is 100 μmol · m −2 s −1 or photosynthetic effective photon as Comparative Example 1 The above-mentioned Production Examples 1A to 1C are used using the cucurbit rhinoceros cultivated under the white fluorescent lamp under the white fluorescent lamp so as to have a bundle density of 100 μmol · m −2 s −1, and It extracted similarly to manufacture, and was set as manufacture examples 2A-7C, comparative manufacture examples 1A-1C, and comparison manufacture examples 2A-2C (Table 2).
実験例１ 活性酸素消去作用
フリーラジカル捕捉除去作用の評価を行った。陽性対照としてはアスコルビン酸を用いた。フリーラジカルのモデルとしては、安定なフリーラジカルであるα，α−ジフェニル−β−ピクリルヒドラジル（以下ＤＰＰＨとする）を用い、試料と一定の割合で一定時間反応させ、減少するラジカルの量を波長５１７ｎｍの吸光度の減少量から測定した。
Experimental Example 1 Reactive Oxygen Scavenging Action The free radical scavenging action was evaluated. Ascorbic acid was used as a positive control. As a model of free radicals, the stable free radical α, α-diphenyl-β-picrylhydrazyl (hereinafter referred to as DPPH) is used, and the amount of radicals which are reacted with the sample at a constant rate for a certain period of time Was determined from the decrease in absorbance at a wavelength of 517 nm.
フリーラジカル捕捉除去作用の測定方法
各試料を、最終濃度０．１〜1．０ｍｇ／ｍＬ（アスコルビン酸は０．０１ｍｇ／ｍＬ）となるように加えた０．１Ｍ酢酸緩衝液（ｐＨ５．５）２ｍＬに無水エタノール２ｍＬ及び０．５ｍＭ ＤＰＰＨ無水エタノール溶液１ｍＬを加えて反応液とした。その後、３７℃で３０分間反応させ、水を対照として波長５１７ｎｍの吸光度を測定した。また、ブランクとして試料の代わりに精製水を加えた反応液を用いて吸光度を測定した。ブランクと比較して、吸光度が５０％減少したときの試料の濃度（ＩＣ５０）を算出した。
Measurement method of free radical scavenging effect 0.1 M acetate buffer (pH 5.5) to which each sample was added to a final concentration of 0.1 to 1.0 mg / mL (ascorbic acid is 0.01 mg / mL) 2 mL of absolute ethanol and 1 mL of 0.5 mM DPPH absolute ethanol solution were added to 2 mL to make a reaction solution. Thereafter, the reaction was allowed to proceed at 37 ° C. for 30 minutes, and the absorbance at a wavelength of 517 nm was measured using water as a control. In addition, the absorbance was measured using a reaction solution to which purified water was added instead of the sample as a blank. The concentration of the sample at which the absorbance decreased by 50% (IC 50 ) was calculated as compared to the blank.
これらの試験結果を表３に示した。本発明の抽出物は、安定で優れたフリーラジカル捕捉除去作用を有していることが認められた。特に、赤色と青色の光量比が４：１〜０：１に高い効果が認められた。その中でも特に、赤色と青色の光量比が３：１〜０：１に高い効果が認められ、３：１に最も高い効果が認められた。なお、アスコルビン酸は、１００℃、１時間の熱処理で失活するが、本発明の抽出物は、活性に変化はなかった。 The test results are shown in Table 3. The extract of the present invention was found to have a stable and excellent free radical scavenging and removing action. In particular, a high effect was observed when the light amount ratio of red to blue was 4: 1 to 0: 1. Among them, particularly, a high effect was observed when the light amount ratio of red to blue was 3: 1 to 0: 1, and the highest effect was observed at 3: 1. Ascorbic acid is inactivated by heat treatment at 100 ° C. for 1 hour, but the extract of the present invention has no change in activity.
実験例２ コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果
Ｉ型コラーゲン（ＣＯＬ１Ａ）及びＭＭＰ−１ ｍＲＮＡ発現量の測定を行った。ヒト皮膚線維芽細胞（ＮＢ１ＲＧＢ）を６０ｍｍ ｄｉｓｈに１×１０５個播種し、１０％ＦＢＳを含むＤＭＥＭ培養液にて、３７℃、５％ＣＯ２条件下で培養した。コンフルエントな状態になったところで、ＣＯＬ１Ａ ｍＲＮＡ発現量測定では各試料を最終濃度１μｇ／ｍＬを添加し、ＭＭＰ−１ ｍＲＮＡ発現量測定では各試料を最終濃度１０μｇ／ｍＬを添加したＤＭＥＭ培養液にて、２４時間培養した後、総ＲＮＡの抽出を行った。細胞からの総ＲＮＡの抽出はＴＲＩＺＯＬ Ｒｅａｇｅｎｔ（Ｉｎｖｉｔｒｏｇｅｎ）を用いて行い、総ＲＮＡ量は分光光度計（ＮａｎｏＤｒｏｐ）を用いて２６０ｎｍにおける吸光度により求めた。ｍＲＮＡ発現量の測定は、細胞から抽出した総ＲＮＡを基にしてリアルタイムＲＴ−ＰＣＲ法により行った。リアルタイムＲＴ−ＰＣＲ法には、ＳｕｐｅｒＳｃｒｉｐｔＩＩＩ Ｐｌａｔｉｎｕｍ Ｔｗｏ−Ｓｔｅｐ ｑＲＴ−ＰＣＲ Ｋｉｔ ｗｉｔｈ ＳＹＢＲ Ｇｒｅｅｎ（Ｉｎｖｉｔｒｏｇｅｎ）を用いた。すなわち、５００ｎｇの総ＲＮＡを逆転写反応後、ＰＣＲ反応（９５℃：１５秒間、６０℃：３０秒間、４０ｃｙｃｌｅｓ）を行った。その他の操作は定められた方法に従い、ＣＯＬ１Ａ及びＭＭＰ−１ ｍＲＮＡの発現量を、内部標準であるβ―ａｃｔｉｎ ｍＲＮＡの発現量に対する割合として求めた。ＣＯＬ１Ａ発現量は、コントロールのＣＯＬ１Ａ ｍＲＮＡの発現量に対する試料添加群のＣＯＬ１Ａ ｍＲＮＡの発現量の比率として算出した。ＭＭＰ発現量についても、同様に算出した。尚、各遺伝子の発現量の測定に使用したプライマーは次の通りである。
Experimental Example 2 Collagen formation promoting effect, matrix metalloproteinase (MMP) inhibitory effect The expression levels of type I collagen (COL1A) and MMP-1 mRNA were measured. Human dermal fibroblasts (NB1 RGB) were seeded at 1 × 10 5 in a 60 mm dish, and cultured in DMEM medium containing 10% FBS under 37 ° C., 5% CO 2 conditions. Once confluent, each sample is added with a final concentration of 1 μg / mL for measurement of COL1A mRNA expression level, and each sample is measured with DMEM culture solution to which a final concentration of 10 μg / mL is added for measurement of MMP-1 mRNA expression level After 24 hours of culture, extraction of total RNA was performed. Extraction of total RNA from cells was performed using TRIZOL Reagent (Invitrogen), and the amount of total RNA was determined by absorbance at 260 nm using a spectrophotometer (NanoDrop). Measurement of mRNA expression level was performed by real-time RT-PCR based on total RNA extracted from cells. SuperScript III Platinum Two-Step qRT-PCR Kit with SYBR Green (Invitrogen) was used for real-time RT-PCR. That is, 500 ng of total RNA was subjected to reverse transcription, followed by PCR (95 ° C .: 15 seconds, 60 ° C .: 30 cycles, 40 cycles). The other procedures were performed according to a prescribed method, and the expression levels of COL1A and MMP-1 mRNA were determined as a percentage of the expression level of β-actin mRNA, which is an internal standard. The COL1A expression level was calculated as a ratio of the expression level of COL1A mRNA of the sample addition group to the expression level of control COL1A mRNA. The MMP expression level was also calculated in the same manner. The primers used to measure the expression level of each gene are as follows.
ＣＯＬ１Ａ用のプライマーセット
ＡＧＧＡＣＡＡＧＡＧＧＣＡＴＧＴＣＴＧＧＴＴ（配列番号１）
ＴＴＧＣＡＧＴＧＧＴＡＧＧＴＧＡＴＧＴＴＣＴＧ（配列番号２）
ＭＭＰ−１用のプライマーセット
ＧＧＧＡＧＡＴＣＡＴＣＧＧＧＡＣＡＡＣＴＣ（配列番号３）
ＴＧＡＧＣＡＴＣＣＣＣＴＣＣＡＡＴＡＣＣ（配列番号４）
β―Ａｃｔｉｎ用のプライマーセット
ＣＡＣＴＣＴＴＣＣＡＧＣＣＴＴＣＣＴＴＣＣ（配列番号５）
ＧＴＧＴＴＧＧＣＧＴＡＣＡＧＧＴＣＴＴＴＧ（配列番号６）
Primer set AGGACAAGAGGCATGTCTGGTT (SEQ ID NO: 1) for COL1A
TTGCAGTGGTAGGTGATGTTCTG (SEQ ID NO: 2)
Primer set for MMP-1 GGGAGATCATCGGGACAACTC (SEQ ID NO: 3)
TGAGCATCCCTCTCAATACC (SEQ ID NO: 4)
Primer set CACTCTTCCAGCCTTCTCTCC (SEQ ID NO: 5) for β-Actin
GTGTTGGCGTACAGGTCTTG (SEQ ID NO: 6)
これらの実験結果を表４、表５に示した。その結果、本発明の抽出物は、優れたＣＯＬ１Ａ発現促進効果（コラーゲン生成促進効果）及びＭＭＰ発現抑制効果（ＭＭＰ阻害効果）が認められた。コラーゲン生成促進効果では、赤色と青色の光量比が８：１〜１：１に高い効果が認められ、その中でも特に、赤色と青色の光量比が４：１〜２：１に高い効果が認められた。ＭＭＰ−１発現抑制効果では、赤色と青色の光量比が８：１〜２：１に高い効果が認められ、その中でも特に、赤色と青色の光量比が４：１に高い効果が認められた。 The results of these experiments are shown in Tables 4 and 5. As a result, the extract of the present invention exhibited an excellent COL1A expression promoting effect (collagen formation promoting effect) and an MMP expression inhibitory effect (MMP inhibitory effect). In the collagen formation promoting effect, a high effect is observed when the light amount ratio of red to blue is 8: 1 to 1: 1, and particularly, a high effect is observed when the light amount ratio of red to blue is 4: 1 to 2: 1 It was done. In the MMP-1 expression suppression effect, a high effect was observed when the light amount ratio of red to blue was 8: 1 to 2: 1, and among them, a high effect was observed when the light amount ratio of red to blue was 4: 1 .
実験例３ メラニン生成抑制試験
対数増殖期にあるＢ１６マウスメラノーマ細胞を６０ｍｍ ｄｉｓｈに３×１０４個播種し、各試料（最終濃度１０μｇ／ｍＬ）を含むＥａｇｌｅｓ’ＭＥＭ（１０％牛胎児血清含有）培地にて、３７℃、５％ＣＯ２条件下で５日間培養した。次に、細胞をｄｉｓｈから剥離し、超音波破砕した後、４Ｎ ＮａＯＨを加え６０℃で２時間の処理を行い、分光光度計でＯ．Ｄ．４７５ｎｍを測定した。尚、超音波処理後の細胞破砕液についてＬｏｗｒｙの方法（Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．，１９３，２６５−２７５，１９５１）にてタンパク定量し、タンパク量当りのメラニン量を算出、試料未添加のメラニン生成量をコントロールとし、コントロールに対する試料添加時のメラニン生成量の値からメラニン生成抑制率を算出した。
Experimental Example 3 Melanin production suppression test 3 × 10 4 B16 mouse melanoma cells in logarithmic growth phase are seeded at 60 mm dish, and each sample (final concentration 10 μg / mL) contains Eagles' MEM (containing 10% fetal calf serum) The medium was cultured at 37 ° C., 5% CO 2 for 5 days. Next, the cells are detached from the dish, sonicated, treated with 4N NaOH at 60 ° C. for 2 hours, and treated with O. D. The 475 nm was measured. The cell disruption solution after sonication is subjected to protein determination by the method of Lowry (J. Biol. Chem., 193, 265-275, 1951), and the amount of melanin per protein amount is calculated. The amount of production was used as a control, and the melanin production suppression rate was calculated from the value of the amount of melanin production at the time of sample addition to the control.
これらの試験結果を表６に示した。本発明の抽出物は、優れたメラニン生成抑制作用を有していることが認められた。特に、赤色と青色の光量比が３：１〜１：１に高い効果が認められた。その中でも特に、赤色と青色の光量比が２：１に高い効果が認められた。 The test results are shown in Table 6. The extract of the present invention was found to have an excellent melanin production inhibitory action. In particular, a high effect was recognized when the light amount ratio of red to blue was 3: 1 to 1: 1. Among them, the light intensity ratio of red to blue was particularly high at 2: 1.
実験例４ 細胞増殖促進試験
ケラチノサイト由来ＨａＣａＴ細胞を９６ｗｅｌｌプレートに１ｗｅｌｌあたり５×１０３個播種し、各試料（最終濃度１μｇ／ｍＬ）を添加した０．１％ＦＢＳを含むＤＭＥＭ培養液にて、３７℃、５％ＣＯ２条件下で３日間培養した。細胞数の測定は、ＭＴＴ法により行った。すなわち、培養終了後、培養液を除き、５００μｇ／ｍＬの濃度にて、ＭＴＴ（３−［４，５−ｄｉｍｅｔｈｙｌｔｈｉａｚｏｌ−２−ｙｌ］−２，５−ｄｉｐｈｅｎｙｌ ｔｅｔｒａｚｏｌｉｕｍ ｂｒｏｍｉｄｅ）を溶解させたＤＭＥＭに培地を入れ替え、２時間培養した後、１５０μＬのｉｓｏｐｒｏｐａｎｏｌに細胞を溶解させ、マイクロプレートリーダーを用いて５７０及び６３０ｎｍにおける吸光度を測定した。細胞数は、５７０ｎｍの吸光度値から、６３０ｎｍの吸光度値を引いた値にて算出し、試料未添加の細胞数をコントロールとし、コントロールに対する試料添加時の細胞数から試料の細胞増殖促進効果を評価した。
Experimental Example 4 Cell Growth Promotion Test Keratinocyte-derived HaCaT cells were seeded at 5 × 10 3 cells / well in a 96-well plate, and each sample (final concentration 1 μg / mL) was added in DMEM culture solution containing 0.1% FBS, The cells were cultured at 37 ° C. under 5% CO 2 for 3 days. Measurement of the cell number was performed by MTT method. That is, after completion of the culture, the culture solution is removed, and the medium is dissolved in DMEM in which MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyl tetrazolium bromide) is dissolved at a concentration of 500 μg / mL. After culturing for 2 hours, cells were dissolved in 150 μl of isopropanol and the absorbance at 570 and 630 nm was measured using a microplate reader. The number of cells is calculated by subtracting the absorbance value of 630 nm from the absorbance value of 570 nm, and using the number of cells to which no sample has been added as a control, evaluates the cell growth promoting effect of the sample did.
これらの実験結果を表７に示した。その結果、本発明の抽出物は、ケラチノサイトに対して優れた細胞増殖促進作用を示した。特に、赤色と青色の光量比が３：１〜１：１に高い効果が認められた。その中でも特に、赤色と青色の光量比が３：１〜２：１に高い効果が認められた。 The results of these experiments are shown in Table 7. As a result, the extract of the present invention showed an excellent cell growth promoting effect on keratinocytes. In particular, a high effect was recognized when the light amount ratio of red to blue was 3: 1 to 1: 1. Among them, a high effect was observed particularly when the light amount ratio of red to blue was 3: 1 to 2: 1.
実験例５ セラミド生成促進試験
セラミド合成における律速酵素遺伝子であるｓｅｒｉｎｅ ｐａｌｍｉｔｏｙｌｔｒａｎｓｆｅｒａｓｅ ｌｏｎｇ ｃｈａｉｎ ｂａｓｅ ｓｕｂｕｎｉｔ １ （ＳＰＴＬＣ１）のｍＲＮＡ発現量を指標として評価した。すなわち、ケラチノサイト由来ＨａＣａＴ細胞を６ｗｅｌｌプレートに１ｗｅｌｌあたり５×１０４個播種し、１０％ＦＢＳを含むＤＭＥＭ培養液にて、３７℃、５％ＣＯ２条件下で４日間培養した。次に、各試料（最終濃度１０μｇ／ｍＬ）を添加したＤＭＥＭ培養液にて、２４時間培養した後、総ＲＮＡの抽出を行った。細胞からの総ＲＮＡの抽出はＴＲＩＺＯＬ Ｒｅａｇｅｎｔ（Ｉｎｖｉｔｒｏｇｅｎ）を用いて行い、総ＲＮＡ量は分光光度計（ＮａｎｏＤｒｏｐ）を用いて２６０ｎｍにおける吸光度により求めた。ｍＲＮＡ発現量の測定は、細胞から抽出した総ＲＮＡを基にしてリアルタイムＲＴ−ＰＣＲ法により行った。リアルタイムＲＴ−ＰＣＲ法には、ＳｕｐｅｒＳｃｒｉｐｔＩＩＩ Ｐｌａｔｉｎｕｍ Ｔｗｏ−Ｓｔｅｐ ｑＲＴ−ＰＣＲ Ｋｉｔ ｗｉｔｈ ＳＹＢＲ Ｇｒｅｅｎ（Ｉｎｖｉｔｒｏｇｅｎ）を用いた。すなわち、５００ｎｇの総ＲＮＡを逆転写反応後、ＰＣＲ反応（９５℃：１５秒間、６０℃：３０秒間、４０ｃｙｃｌｅｓ）を行った。その他の操作は定められた方法に従い、ＳＰＴＬＣ１ ｍＲＮＡの発現量を、内部標準であるＧＡＰＤＨ ｍＲＮＡの発現量に対する割合として求めた。ＳＰＴＬＣ１発現量は、コントロールのＳＰＴＬＣ１ ｍＲＮＡの発現量に対する試料添加群のＳＰＴＬＣ１ ｍＲＮＡの発現量の比率として算出した。尚、各遺伝子の発現量の測定に使用したプライマーは次の通りである。
Experimental Example 5 Ceramide Formation Promotion Test The amount of mRNA expression of serine palmitoyltransferase long chain base subunit 1 (SPTLC1), which is a rate-limiting enzyme gene in ceramide synthesis, was evaluated as an index. That is, keratinocyte-derived HaCaT cells were seeded at 5 × 10 4 cells / well in a 6-well plate, and cultured in a DMEM culture solution containing 10% FBS at 37 ° C. under 5% CO 2 for 4 days. Next, after culturing for 24 hours in a DMEM culture solution to which each sample (final concentration 10 μg / mL) was added, extraction of total RNA was performed. Extraction of total RNA from cells was performed using TRIZOL Reagent (Invitrogen), and the amount of total RNA was determined by absorbance at 260 nm using a spectrophotometer (NanoDrop). Measurement of mRNA expression level was performed by real-time RT-PCR based on total RNA extracted from cells. SuperScript III Platinum Two-Step qRT-PCR Kit with SYBR Green (Invitrogen) was used for real-time RT-PCR. That is, 500 ng of total RNA was subjected to reverse transcription, followed by PCR (95 ° C .: 15 seconds, 60 ° C .: 30 cycles, 40 cycles). Other procedures were performed according to a defined method, and the expression level of SPTLC1 mRNA was determined as a ratio to the expression level of GAPDH mRNA which is an internal standard. The SPTLC1 expression level was calculated as the ratio of the expression level of SPTLC1 mRNA of the sample addition group to the expression level of SPTLC1 mRNA of control. The primers used to measure the expression level of each gene are as follows.
ＳＰＴＬＣ１用のプライマーセット
ＴＧＧＴＣＡＣＧＧＴＧＧＡＡＣＡＡＡＣＡ（配列番号７）
ＧＣＣＴＧＧＧＣＴＡＣＣＴＣＣＴＴＧＡ（配列番号８）
ＧＡＰＤＨ用のプライマーセット
ＴＧＣＡＣＣＡＣＣＡＡＣＴＧＣＴＴＡＧＣ（配列番号９）
ＴＣＴＴＣＴＧＧＧＴＧＧＣＡＧＴＧＡＴＧ（配列番号１０）
Primer set TGGTCACGGTGGAACAAACA (SEQ ID NO: 7) for SPTLC1
GCCTGGGCTACCTCCTTGA (SEQ ID NO: 8)
Primer set TGCACCACCAACTGCTTAGC (SEQ ID NO: 9) for GAPDH
TCTTCTGGGTGGCAGTGATG (SEQ ID NO: 10)
これらの実験結果を表８に示した。その結果、本発明の抽出物はセラミド生成促進効果を示した。赤色と青色の光量比が８：１〜１：１に高い効果が認められた。その中でも、赤色と青色の光量比が４：１〜２：１に高い効果が認められ４：１〜３：１に最も高い効果が認められた。 The results of these experiments are shown in Table 8. As a result, the extract of the present invention showed a ceramide formation promoting effect. A high effect was recognized when the light amount ratio of red to blue was 8: 1 to 1: 1. Among them, a high effect is observed when the light amount ratio of red to blue is 4: 1 to 2: 1, and the highest effect is observed between 4: 1 to 3: 1.
処方例１ 化粧水
処方 配合量（部）
１．製造例３Ａの抽出物 １．０
２．１，３‐ブチレングリコール ８．０
３．グリセリン ２．０
４．キサンタンガム ０．０２
５．クエン酸 ０．０１
６．クエン酸ナトリウム ０．１
７．エタノール ５．０
８．パラオキシ安息香酸メチル ０．１
９．ポリオキシエチレン硬化ヒマシ油（４０Ｅ．Ｏ．） ０．１
１０．香料 適量
１１．精製水にて全量を１００とする
［製造方法］成分１〜６及び１１と、成分７〜１０をそれぞれ均一に溶解し、両者を混合し濾過して製品とする。
Prescription example 1 Lotion prescription quantity (part)
1. Extract of Preparation Example 3A 1.0
2. 1, 3- butylene glycol 8.0
3. Glycerin 2.0
4. Xanthan gum 0.02
5. Citric acid 0.01
6. Sodium citrate 0.1
7. Ethanol 5.0
8. Methyl parahydroxybenzoate 0.1
9. Polyoxyethylene Cured Castor Oil (40E.O.) 0.1
10. Perfume amount 11. [Production method] The total amount is made 100 with purified water. Components 1 to 6 and 11 and components 7 to 10 are uniformly dissolved, and both are mixed and filtered to obtain a product.
処方例２ クリーム
処方 配合量（部）
１．製造例４Ａの抽出物 ０．５
２．スクワラン ５．５
３．オリーブ油 ３．０
４．ステアリン酸 ２．０
５．ミツロウ ２．０
６．ミリスチン酸オクチルドデシル ３．５
７．ポリオキシエチレンセチルエーテル（２０Ｅ．Ｏ．） ３．０
８．ベヘニルアルコール １．５
９．モノステアリン酸グリセリン ２．５
１０．香料 ０．１
１１．パラオキシ安息香酸メチル ０．２５
１２．１，３‐ブチレングリコール ８．５
１３．精製水にて全量を１００とする
［製造方法］成分２〜９を加熱溶解して混合し、７０℃に保ち油相とする。成分１及び１１〜１３を加熱溶解して混合し、７５℃に保ち水相とする。油相に水相を加えて乳化して、かき混ぜながら冷却し、４５℃で成分１０を加え、更に３０℃まで冷却して製品とする。
Prescription example 2 Cream prescription formula amount (part)
1. Extract of Production Example 4A 0.5
2. Squalane 5.5
3. Olive oil 3.0
4. Stearic acid 2.0
5. Bee wax 2.0
6. Octyldodecyl myristate 3.5
7. Polyoxyethylene cetyl ether (20E.O.) 3.0
8. Behenyl alcohol 1.5
9. Glycerin monostearate 2.5
10. Flavoring agent 0.1
11. Methyl parahydroxybenzoate 0.25
12. 1,3-butylene glycol 8.5
13. The total amount is made 100 with purified water. [Production method] Ingredients 2 to 9 are dissolved by heating, mixed, and kept at 70 ° C. to form an oil phase. Ingredients 1 and 11 to 13 are dissolved by heating, mixed and kept at 75 ° C. to form an aqueous phase. Add the aqueous phase to the oil phase and emulsify, cool with stirring, add ingredient 10 at 45 ° C, and further cool to 30 ° C to give a product.
処方例２において、製造例４Ａの抽出物を製造例１Ａの抽出物、製造例７Ａの抽出物及び製造例４Ｂの抽出物に置き換えたものを処方例３、４及び５とした。 In Formulation Example 2, Formulations 3, 4 and 5 were prepared by replacing the extract of Production Example 4A with the extract of Production Example 1A, the extract of Production Example 7A and the extract of Production Example 4B.
処方例６ 乳液
処方 配合量（部）
１．製造例２Ａの抽出物 １．０
２．スクワラン ５．０
３．オリーブ油 ５．０
４．ホホバ油 ５．０
５．セタノール １．５
６．モノステアリン酸グリセリン ２．０
７．ポリオキシエチレンセチルエーテル（２０Ｅ．Ｏ．） ３．０
８．ポリオキシエチレンソルビタンモノオレエート（２０Ｅ．Ｏ．） ２．０
９．香料 ０．１
１０．プロピレングリコール １．０
１１．グリセリン ２．０
１２．パラオキシ安息香酸メチル ０．２
１３．精製水にて全量を１００とする
［製造方法］成分２〜８を加熱溶解して混合し、７０℃に保ち油相とする。成分１及び１０〜１３を加熱溶解して混合し、７５℃に保ち水相とする。油相に水相を加えて乳化して、かき混ぜながら冷却し、４５℃で成分９を加え、更に３０℃まで冷却して製品とする。
Prescription example 6 Emulsion prescription quantity (part)
1. Extract of Production Example 2A 1.0
2. Squalane 5.0
3. Olive oil 5.0
4. Jojoba oil 5.0
5. Settanol 1.5
6. Glycerin monostearate 2.0
7. Polyoxyethylene cetyl ether (20E.O.) 3.0
8. Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Flavoring agent 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12. Methyl parahydroxybenzoate 0.2
13. The total amount is made 100 with purified water. [Production method] Ingredients 2 to 8 are dissolved by heating, mixed, and kept at 70 ° C. to form an oil phase. Ingredients 1 and 10 to 13 are dissolved by heating, mixed and kept at 75 ° C. to form an aqueous phase. The water phase is added to the oil phase and emulsified, cooled with stirring, added with component 9 at 45 ° C., and further cooled to 30 ° C. to give a product.
処方例７ ゲル剤
処方 配合量（部）
１．製造例４Ｃの抽出物 ０．００１
２．エタノール ５．０
３．パラオキシ安息香酸メチル ０．１
４．ポリオキシエチレン硬化ヒマシ油（６０Ｅ．Ｏ．） ０．１
５．香料 適量
６．１，３‐ブチレングリコール ５．０
７．グリセリン ５．０
８．キサンタンガム ０．１
９．カルボキシビニルポリマー ０．２
１０．水酸化カリウム ０．２
１１．精製水にて全量を１００とする
［製造方法］成分２〜５と、成分１及び６〜１１をそれぞれ均一に溶解し、両者を混合して製品とする。
Formulation example 7 Gel formulation Formulation amount (parts)
1. Extract of Preparation Example 4C 0.001
2. Ethanol 5.0
3. Methyl parahydroxybenzoate 0.1
4. Polyoxyethylene Cured Castor Oil (60E.O.) 0.1
5. Fragrance appropriate amount 6.1, 3- butylene glycol 5.0
7. Glycerin 5.0
8. Xanthan gum 0.1
9. Carboxy vinyl polymer 0.2
10. Potassium hydroxide 0.2
11. The total amount is made 100 with purified water. [Production method] Ingredients 2 to 5 and ingredients 1 and 6 to 11 are uniformly dissolved, and both are mixed to make a product.
処方例８ パック
処方 配合量（部）
１．製造例５Ａの抽出物 ０．１
２．製造例５Ｂの抽出物 ０．１
３．ポリビニルアルコール １２．０
４．エタノール ５．０
５．１，３‐ブチレングリコール ８．０
６．パラオキシ安息香酸メチル ０．２
７．ポリオキシエチレン硬化ヒマシ油（２０Ｅ．Ｏ．） ０．５
８．クエン酸 ０．１
９．クエン酸ナトリウム ０．３
１０．香料 適量
１１．精製水にて全量を１００とする
［製造方法］成分１〜１１を均一に溶解し製品とする。
Prescription example 8 Pack prescription combination amount (part)
1. Extract of Preparation Example 5A 0.1
2. Extract of Preparation Example 5B 0.1
3. Polyvinyl alcohol 12.0
4. Ethanol 5.0
5.1,3-Butylene glycol 8.0
6. Methyl parahydroxybenzoate 0.2
7. Polyoxyethylene Cured Castor Oil (20E.O.) 0.5
8. Citric acid 0.1
9. Sodium citrate 0.3
10. Perfume amount 11. [Manufacturing method] the total amount is made 100 with purified water. Components 1 to 11 are uniformly dissolved to obtain a product.
処方例９ ファンデーション
処方 配合量（部）
１．製造例３Ａの抽出物 １．０
２．ステアリン酸 ２．４
３．ポリオキシエチレンソルビタンモノステアレート（２０Ｅ．Ｏ．） １．０
４．ポリオキシエチレンセチルエーテル（２０Ｅ．Ｏ．） ２．０
５．セタノール １．０
６．液状ラノリン ２．０
７．流動パラフィン ３．０
８．ミリスチン酸イソプロピル ６．５
９．カルボキシメチルセルロースナトリウム ０．１
１０．ベントナイト ０．５
１１．プロピレングリコール ４．０
１２．トリエタノールアミン １．１
１３．パラオキシ安息香酸メチル ０．２
１４．二酸化チタン ８．０
１５．タルク ４．０
１６．ベンガラ １．０
１７．黄酸化鉄 ２．０
１８．香料 適量
１９．精製水にて全量を１００とする
［製造方法］成分２〜８を加熱溶解し、８０℃に保ち油相とする。成分１９に成分９をよく膨潤させ、続いて、成分１及び１０〜１３を加えて均一に混合する。これに粉砕機で粉砕混合した成分１４〜１７を加え、ホモミキサーで撹拌し７５℃に保ち水相とする。この油相に水相をかき混ぜながら加え、乳化する。その後冷却し、４５℃で成分１８を加え、かき混ぜながら３０℃まで冷却して製品とする。
Prescription example 9 Foundation Prescription Formulation amount (parts)
1. Extract of Preparation Example 3A 1.0
2. Stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate (20E.O.) 1.0
4. Polyoxyethylene cetyl ether (20E.O.) 2.0
5. Settanol 1.0
6. Liquid lanolin 2.0
7. Liquid paraffin 3.0
8. Isopropyl myristate 6.5
9. Carboxymethylcellulose sodium 0.1
10. Bentonite 0.5
11. Propylene glycol 4.0
12. Triethanolamine 1.1
13. Methyl parahydroxybenzoate 0.2
14. Titanium dioxide 8.0
15. Talc 4.0
16. Bengala 1.0
17. Yellow iron oxide 2.0
18. Perfume amount 19. The total amount is made 100 with purified water. [Production method] Ingredients 2 to 8 are dissolved by heating and kept at 80 ° C. to form an oil phase. Allow component 19 to swell well, then add components 1 and 10-13 and mix uniformly. The ingredients 14-17 pulverized and mixed with a grinder are added to this, it stirs with a homomixer, and it keeps at 75 degreeC, and sets it as an aqueous phase. The water phase is added to this oil phase while stirring and emulsifying. It is then cooled, component 18 is added at 45 ° C. and cooled to 30 ° C. with stirring to give a product.
処方例１０ 浴用剤
処方 配合量（部）
１．製造例６Ａの抽出物 ５．０
２．製造例６Ｂの抽出物 １．０
３．炭酸水素ナトリウム ５０．０
４．黄色２０２号（１） 適量
５．香料 適量
６．硫酸ナトリウムにて全量を１００とする
［製造方法］成分１〜６を均一に混合し製品とする。
Formulation example 10 Bath preparation Formulation amount (parts)
1. Extract of Production Example 6A 5.0
2. Extract of Preparation Example 6B 1.0
3. Sodium bicarbonate 50.0
4. Yellow No. 202 (1) Appropriate amount 5. Flavoring amount 6. The total amount is made 100 with sodium sulfate. [Production method] Ingredients 1 to 6 are uniformly mixed to obtain a product.
処方例１１ 軟膏
処方 配合量（部）
１．製造例４Ｂの抽出物 ０．５
２．ポリオキシエチレンセチルエーテル（３０Ｅ．Ｏ．） ２．０
３．モノステアリン酸グリセリン １０．０
４．流動パラフィン ５．０
５．セタノール ６．０
６．パラオキシ安息香酸メチル ０．１
７．プロピレングリコール １０．０
８．精製水にて全量を１００とする
［製造方法］成分２〜５を加熱溶解して混合し、７０℃に保ち油相とする。成分１及び６〜８を加熱溶解して混合し、７５℃に保ち水相とする。油相に水相を加えて乳化して、かき混ぜながら３０℃まで冷却して製品とする。
Prescription example 11 Ointment Prescription combination amount (part)
1. Extract of Production Example 4B 0.5
2. Polyoxyethylene cetyl ether (30E.O.) 2.0
3. Glycerin monostearate 10.0
4. Liquid paraffin 5.0
5. Setanol 6.0
6. Methyl parahydroxybenzoate 0.1
7. Propylene glycol 10.0
8. The total amount is made 100 with purified water. [Production method] Ingredients 2 to 5 are dissolved by heating, mixed, and kept at 70 ° C. to form an oil phase. Ingredients 1 and 6 to 8 are heated, dissolved and mixed, kept at 75 ° C. to form an aqueous phase. The water phase is added to the oil phase, emulsified, and cooled to 30 ° C. while stirring to give a product.
処方例１２ 散剤
処方 配合量（部）
１．製造例４Ａの抽出物 ２０．０
２．乾燥コーンスターチ ３０．０
３．微結晶セルロース ５０．０
［製造方法］成分１〜３を混合し、散剤とする。
Formulation example 12 Powder formulation Formulation amount (parts)
1. Extract of Preparation Example 4A 20.0
2. Dried corn starch 30.0
3. Microcrystalline cellulose 50.0
[Production method] Ingredients 1 to 3 are mixed to make a powder.
処方例１３ 錠剤
処方 配合量（部）
１．製造例４Ａの抽出物 ３．０
２．乾燥コーンスターチ ２７．０
３．カルボキシメチルセルロースカルシウム ２０．０
４．微結晶セルロース ４０．０
５．ポリビニルピロリドン ７．０
６．タルク ３．０
［製造方法］成分１〜４を混合し、次いで成分５の水溶液を結合剤として加えて顆粒成形する。成形した顆粒に成分６を加えて打錠する。１錠０．５２ｇとする。
Prescription Example 13 Tablet Prescription Amount (parts)
1. Extract of Preparation Example 4A 3.0
2. Dried corn starch 27.0
3. Carboxymethylcellulose calcium 20.0
4. Microcrystalline cellulose 40.0
5. Polyvinylpyrrolidone 7.0
6. Talc 3.0
[Production method] Ingredients 1 to 4 are mixed, then an aqueous solution of ingredient 5 is added as a binder and granulated. Ingredient 6 is added to the shaped granules and compressed. It will be 0.52 g of 1 tablet.
処方例１４ 錠菓
処方 配合量（部）
１．製造例５Ａの抽出物 ０．５
２．乾燥コーンスターチ ５０．０
３．エリスリトール ４０．０
４．クエン酸 ５．０
５．ショ糖脂肪酸エステル ３．０
６．香料 適量
７．精製水にて全量を１００とする
［製造方法］成分１〜４及び７を混合し、顆粒成形する。成形した顆粒に成分５及び６を加えて打錠する。１粒１．０ｇとする。
Prescription example 14 Tablet confectionery Prescription combination amount (part)
1. Extract of Production Example 5A 0.5
2. Dried corn starch 50.0
3. Erythritol 40.0
4. Citric acid 5.0
5. Sucrose fatty acid ester 3.0
6. Perfume amount 7. Make up the whole amount to 100 with purified water [Production method] Ingredients 1 to 4 and 7 are mixed and granulated. Ingredients 5 and 6 are added to the shaped granules and compressed. It is 1 grain 1.0g.
処方例１５ 飲料
処方 配合量（部）
１．製造例５Ａの抽出物 ２．０
２．果糖ブドウ糖液糖 １２．５
３．クエン酸 ０．1
４．香料 ０．０５
５．精製水にて全量を１００とする
［製造方法］成分１〜５を混合し、飲料とする。
Prescription example 15 Beverage Prescription Formulation amount (parts)
1. Extract of Preparation Example 5A 2.0
2. Fructose glucose liquid sugar 12.5
3. Citric acid 0.1
4. Fragrance 0.05
5. Make 100 parts with purified water [Production method] Ingredients 1 to 5 are mixed to make a beverage.
処方例１６ 粉末飲料
処方 配合量（部）
１．製造例７Ａの抽出物 １０．０
２．粉糖 ６５．０
３．粉末ピーチ果汁 １５．０
４．Ｌ−アスコルビン酸 ８．０
５．結晶クエン酸 １．２
６．クエン酸ナトリウム ０．７５
７．アスパルテーム ０．０２
８．粉末ピーチ香料 ０．０３
［製造方法］成分１〜８を混合し、粉末飲料とする。
Formulation example 16 Powdered beverage Formulation amount (parts)
1. Extract of Preparation Example 7A 10.0
2. Powdered sugar 65.0
3. Powdered peach juice 15.0
4. L-ascorbic acid 8.0
5. Crystalline citric acid 1.2
6. Sodium citrate 0.75
7. Aspartame 0.02
8. Powdered peach flavor 0.03
[Production method] Ingredients 1 to 8 are mixed to make a powdered beverage.
処方例１７ ハーブティ
処方 配合量（部）
１．クウシンサイ乾燥物（比較例を除く実施例１の表１） １．０
２．ペパーミント ０．５
３．ローズヒップ ０．５
［製造方法］成分１〜３を混合し、ティーバッグに２ｇを封入してハーブティーとする。
Prescription Example 17 Herbal Tea Prescription Amount (parts)
1. Dried Kuusinsai (Table 1 of Example 1 excluding Comparative Example) 1.0
2. Peppermint 0.5
3. Rose hip 0.5
[Production method] Ingredients 1 to 3 are mixed, and 2 g is enclosed in a tea bag to make herbal tea.
以上のことから、特定の波長域を有する光を照射して栽培したクウシンサイやその抽出物は、優れた抗酸化効果、コラーゲン生成促進効果、マトリックスメタロプロテアーゼ（ＭＭＰ）阻害効果、美白効果、細胞増殖効果、セラミド産生促進効果を示し、これらを含有する皮膚外用剤又は内用剤は特に有効である。 From the above, it has been found that the kushishinsai cultivated by irradiation with light having a specific wavelength range and its extract have excellent antioxidative effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibitory effect, whitening effect, cell proliferation The effect, ceramide production promoting effect is exhibited, and a skin external preparation or an internal preparation containing them is particularly effective.
Claims (3)
Compared with the Chinese cabbage grown with fluorescent light or sunlight, by cultivating by irradiating with light with a photosynthetic photon flux density (PPFD) ratio of 4: 1 to 2: 1 between the wavelength range of 570 to 730 nm and 400 to 515 nm. An aqueous solution of one or more selected from antioxidative effect, collagen generation promoting effect, matrix metalloproteinase (MMP) inhibiting effect, whitening effect, cell proliferation effect and ceramide production promoting effect, or water, low grade A food comprising an extract extracted by one or more solvents selected from alcohol and liquid polyhydric alcohol.
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