JP5259127B2 - A composition for an integumental tissue containing at least a component derived from a plant belonging to the genus Ericaceae - Google Patents
A composition for an integumental tissue containing at least a component derived from a plant belonging to the genus Ericaceae Download PDFInfo
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Description
本発明は、外皮系組織産生に関わる技術に関する。より詳しくは、ツツジ科エリカ属植物由来成分を少なくとも含有する外皮系組織産生促進用組成物、並びに該組成物を有効成分として含有する医薬品、飲食物、及び化粧品に関する。 The present invention relates to a technique related to the production of integumentary tissue. More specifically, the present invention relates to a composition for promoting the production of an outer skin tissue containing at least a component derived from a plant belonging to the genus Eridaceae, and a pharmaceutical, a food and drink, and a cosmetic containing the composition as an active ingredient.
外皮とは、体表を覆う皮膚と、毛、爪、汗腺及び脂腺など皮膚の表皮の変形物との総称である。外皮系組織は、ヒト又は動物の最も外側にある組織であり、保護機能、保温機能、感覚機能などの多様な機能を有し、ヒト及び動物の生存には不可欠なものである。また、体内の変化を示す重要な組織でもある。例えば、体内のビリルビン過剰による黄疸、ストレスによる脱毛、動物の毛艶の変化など、異変を確認し易い組織でもある。更に、特にヒトにおいては、外観的な美的印象を左右する組織でもある。 The outer skin is a general term for skin covering the body surface and skin epidermal deformations such as hair, nails, sweat glands and sebaceous glands. The integumentary tissue is the outermost tissue of a human or animal, has various functions such as a protective function, a heat retaining function, and a sensory function, and is indispensable for the survival of humans and animals. It is also an important tissue that shows changes in the body. For example, it is also a tissue that is easy to confirm anomalies such as jaundice due to excessive bilirubin in the body, hair loss due to stress, and changes in animal gloss. Furthermore, especially in humans, it is also an organization that influences the appearance and aesthetic impression.
外皮系組織の一つであるヒト又は動物の皮膚を覆う毛は、皮膚の角化細胞が分化してできた角質性の付属物である。 Hair that covers human or animal skin, which is one of the integumentary tissues, is a keratinous appendage formed by differentiation of keratinocytes of the skin.
毛（毛軸）は、ヘアマトリックス（毛母基）における毛包構成角化細胞の増殖により成長する。一定期間成長した後（成長期）、毛包と毛乳頭の退化が起こり（退行期）、やがて毛軸は棍棒状（棍毛）になり立毛筋の付着する毛包バルジ領域付近より上部にとどまるか、若しくは脱落する（休止期）。しばらくすると、残存する外毛根鞘の下端部に、新たに毛乳頭とヘアマトリックス（毛母基）が形成され、再びヘアマトリックス（毛母基）における毛包構成角化細胞の分裂が開始し、成長期へと入るという毛周期を繰り返す。正常な場合、一定の毛周期によって、成長する毛と抜け落ちる毛のバランスが保たれている。 Hair (hair shaft) grows by proliferation of hair follicle-forming keratinocytes in the hair matrix (hair matrix). After growing for a certain period (growth stage), degeneration of the hair follicle and dermal papilla occurs (regression stage), eventually the hair shaft becomes a rod-shaped (lash) and stays above the vicinity of the hair follicle bulge area where the napped muscles adhere Or drop out (rest period). After a while, a new hair papilla and hair matrix (hair matrix) are formed at the lower end of the remaining outer root sheath, and division of hair follicle keratinocytes in the hair matrix (hair matrix) starts again. Repeat the hair cycle of entering the growth period. Under normal conditions, a constant hair cycle balances the growing hair and the falling hair.
しかし、何らかの原因でヘアマトリックス（毛母基）や毛乳頭に障害を受けると、前記毛周期サイクルが乱れ、成長期から退行期、休止期へと急激に移行し、脱毛が起こる。この原因は、様々であるが、例えば、老化、ホルモンバランスの乱れ、自己免疫異常、精神的ストレス、環境変化、薬物の副作用などが挙げられる。 However, when the hair matrix (hair matrix) or the hair papilla is damaged for some reason, the hair cycle cycle is disturbed, and the hair transitions rapidly from the growth phase to the regression phase and the resting phase, and hair loss occurs. There are various causes for this, and examples include aging, disorder of hormone balance, autoimmune abnormality, mental stress, environmental changes, and side effects of drugs.
近年、上記脱毛を予防又は治療するための、様々な薬剤等が開発されている。例えば、特許文献１には、塩化カルプロニウムまたはその水和物、およびミノキシジルまたはその塩を含有した発毛用組成物が開示されている。また、特許文献２には、テストステロン５α−リダクターゼ阻害剤と、アデノシン等のアデノシン類とを含有する頭皮頭髪用組成物が開示されている。 In recent years, various drugs and the like have been developed for preventing or treating hair loss. For example, Patent Document 1 discloses a hair growth composition containing carpronium chloride or a hydrate thereof and minoxidil or a salt thereof. Patent Document 2 discloses a composition for scalp and hair containing a testosterone 5α-reductase inhibitor and adenosines such as adenosine.
外皮系組織の一つである皮膚や毛には、過剰な光を吸収し、紫外線を遮る役割を担うメラニンが存在する。メラニンは、皮膚、毛、眼球などのメラノサイトで合成される褐色ないし黒色の色素であり、隣接細胞に拡散することで、日焼け、シミ、ソバカスなどの色素沈着を起こすが、その一方で、紫外線や電離性放射線から人体を守るといったバリヤーの役割を担う重要な色素でもある。 In skin and hair, which is one of the integumental tissues, there is melanin that plays a role of absorbing excessive light and blocking ultraviolet rays. Melanin is a brown or black pigment synthesized in melanocytes such as skin, hair, and eyeballs, and when it diffuses into neighboring cells, it causes pigmentation such as sunburn, spots, and freckles. It is also an important pigment that plays the role of a barrier to protect the human body from ionizing radiation.
メラニンは、メラノサイトの中の小器官であるメラノソームに存在するチロシナーゼの作用によってチロシンから生じるドパ（ＤＯＰＡ）が酸化、脱炭酸を経て産生されるドパクロムやインドールキノンなどが重合して合成される。 Melanin is synthesized by polymerizing dopachrome, indolequinone, etc. produced by oxidation and decarboxylation of dopa (DOPA) generated from tyrosine by the action of tyrosinase present in melanosomes, which are organelles in melanocytes.
メラニンの産生が異常に抑制されてしまうと、毛髪や皮膚および目の網膜の色素異常が生じる。例えば、毛髪中のメラニンが欠乏すると白髪が、皮膚中のメラニンは欠乏すると尋常性白斑等の色素脱失症が生じる。また、メラニンの合成の重要な酵素であるチロシナーゼが欠損すると、メラニン合成障害により、白皮症を生じる。 If the production of melanin is abnormally suppressed, pigment abnormalities in the hair, skin and eye retina will occur. For example, deficiency of melanin in hair causes white hair, and deficiency of melanin in skin causes depigmentation such as vulgaris. In addition, deficiency of tyrosinase, which is an important enzyme for melanin synthesis, causes albinism due to melanin synthesis disorder.
メラニン産生に関する技術として、引用文献３には、Ｒａｎｕｎｃｕｌａｃｅａｅ（キンポウゲ科）Ｎｉｇｅｌｌａ（クロタネソウ）属植物あるいはその抽出物を有効成分として含有する毛髪用のメラニン産生促進剤が、引用文献４には、ネジトウガラシ(Helicteres isora L.,Sterculiaceae）、クミスクチン(Orthosiphon aristatus Bl.）、カユプテ（Melaleuca leucadendra L.）及びこれらの抽出物のうちの少なくとも１種が有効成分として配合されてなることを特徴とするチロシナーゼ生合成促進剤が開示されている。 As a technique relating to melanin production, Cited Document 3 discloses a melanin production promoter for hair containing a plant belonging to the genus Ranunculaceae (Nymphaceae) Nigella (Polygonaceae) or an extract thereof as an active ingredient, and Cited Document 4 discloses a screw pepper. (Helicteres isora L., Sterculiaceae), cumistin (Orthosiphon aristatus Bl.), Cayupute (Melaleuca leucadendra L.) and at least one of these extracts as an active ingredient. Synthesis accelerators are disclosed.
ここで、本発明に関連のあるツツジ科エリカ属植物について、以下説明する。ツツジ科エリカ属植物は、その多くが南アフリカ原産の高さ０．２から１．５ｍ程度の低木であるが、中には、高さ６から７ｍに達する種もある。ツツジ科エリカ属植物は、ハーブティの材料として、その葉、花、茎などが使われたり、入浴剤として使用されたりすることが多い。また、様々な疾患予防又は治療に役立つことが報告されている。 Here, Erythaceae genus plants related to the present invention will be described below. Most of the Euraceae genus plants are shrubs with a height of about 0.2 to 1.5 m native to South Africa, but some species reach a height of 6 to 7 m. The azalea family Erika is often used as a herbal tea, its leaves, flowers, stems, etc., or as a bath. It has also been reported to be useful for the prevention or treatment of various diseases.
エリカを用いた薬剤として、引用文献５には、コラゲナーゼ抑制効果を有する薬剤が、引用文献６には抗血栓剤にエリカを用いることが開示されている。更に、引用文献７には、毛髪を脱色に使用する化粧料にツツジ科エリカ属植物のエッセンスを含有させることが開示されている。 As a drug using Erica, Cited Document 5 discloses a drug having a collagenase inhibitory effect, and Cited Document 6 discloses using Erica as an antithrombotic agent. Furthermore, cited document 7 discloses that the essence of the genus Ericaceae is contained in a cosmetic used for decolorizing hair.
外皮系組織に関わる疾患は、原因が不明であったり、治療が困難であったりする場合が多い。また、環境の変化、精神的ストレスなどの影響により、近年、年齢に関わらず、外皮系組織に関わる疾患の罹患が増加している。そのため、より安全な方法でかつ、長期にわたり継続できる治療方法が望まれる。 In many cases, diseases associated with the integumental tissue have an unknown cause or are difficult to treat. In recent years, due to the influence of environmental changes, mental stress, etc., the incidence of diseases related to the integumental tissue is increasing regardless of age. Therefore, a safer method and a treatment method that can be continued for a long time are desired.
そこで、本発明は、外皮系組織産生促進効果を有し、安全性の高い組成物を提供することを主目的とする。 Therefore, the main object of the present invention is to provide a highly safe composition that has an effect of promoting the production of outer skin tissue.
本願発明者は、上記目的を達成するために、種々の天然物を探索した。その結果、ツツジ科エリカ属植物由来成分が、外皮系組織産生促進効果を有することを新規に見出した。 The inventor of the present application searched for various natural products in order to achieve the above object. As a result, it has been newly found out that a component derived from a plant belonging to the genus Ericaceae has an effect of promoting the production of outer skin tissue.
まず、本発明では、ツツジ科エリカ属植物由来成分を少なくとも含有する外皮系組織産生促進用組成物を提供する。本発明に係る外皮系組織産生促進用組成物は、ツツジ科エリカ属植物由来成分を少なくとも含有すれば、該植物の株種は特に限定されない。好適な一例としては、Ｅｒｉｃａ ｍｕｌｔｉｆｌｏｒａが挙げられる。 First, the present invention provides a composition for promoting the production of integumental tissue, which contains at least a component derived from a plant belonging to the genus Ericaceae. As long as the composition for promoting skin tissue production according to the present invention contains at least a component derived from the genus Ericaceae, the plant species of the plant is not particularly limited. A suitable example is Erica multiflora.
本発明に係る外皮系組織産生促進用組成物は、外皮系組織産生を促す種々の作用を有する。具体的な作用の一例としては、発毛促進作用、毛乳頭細胞増殖促進作用、毛周期成長期誘導作用、血管拡張作用、メラニン産生促進作用及びチロシナーゼ合成促進作用などが挙げられる。 The composition for promoting the production of the outer skin tissue according to the present invention has various actions for promoting the production of the outer skin tissue. Specific examples of the action include a hair growth promoting action, a hair papillary cell proliferation promoting action, a hair cycle growth phase inducing action, a vasodilatory action, a melanin production promoting action, and a tyrosinase synthesis promoting action.
本発明に係る外皮系組織産生促進用組成物は、これを有効成分として、医薬品、飲食物、及び化粧品に適用することができる。 The composition for promoting the production of the outer skin tissue according to the present invention can be applied to pharmaceuticals, foods and drinks, and cosmetics using this as an active ingredient.
ここで、本発明で使用する技術用語の定義付けを行う。本発明における「外皮系組織」とは、表皮、真皮、及び皮下組織に存在する全ての組織を含む。例えば、皮膚、毛、爪、汗腺・皮脂腺・乳腺等の付属皮膚腺、肉球、蹄、角など、更に、これらの組織に含まれる細胞、色素、血管等も含む。「毛」とは、ヒト又は動物の皮膚を覆う毛の全てを意味し、体毛、毛髪など、全ての毛を包含する。「発毛」とは、毛の成長を促すことの全てを意味し、毛周期における休止期の状態から成長期へと誘導して再び「発毛」させること、及び既に生えている毛の成長を促す「育毛」という概念全てを包含する。 Here, technical terms used in the present invention are defined. In the present invention, “dermis tissue” includes all tissues present in the epidermis, dermis, and subcutaneous tissue. For example, skin, hair, nails, accessory skin glands such as sweat glands, sebaceous glands, mammary glands, paws, hoofs, horns, etc., and cells, pigments, blood vessels, etc. contained in these tissues are also included. “Hair” means all hair covering the skin of a human or animal and includes all hair such as body hair and hair. “Growing” means all of promoting hair growth, leading from the resting state in the hair cycle to the growing phase to “grow” again, and growing hair that has already grown Includes all the concepts of “hair growth” that promotes hair growth.
本発明に係る組成物は、外皮系組織産生促進効果を有する。また、本発明に係る組成物は、天然由来成分であり、飲食物として摂取されたり、化粧品として利用されたりした実績があるため、長期に連続的に使用しても安全である可能性が高い。 The composition according to the present invention has an effect of promoting the production of outer skin tissue. In addition, the composition according to the present invention is a naturally-derived component and has a track record of being consumed as a food or drink or used as a cosmetic, so it is highly likely that it is safe to use continuously for a long period of time. .
以下、本発明を実施するための好適な形態について説明する。なお、以下に説明する実施形態は、本発明の代表的な実施形態の一例を示したものであり、これにより本発明の範囲が狭く解釈されることはない。 Hereinafter, preferred embodiments for carrying out the present invention will be described. In addition, embodiment described below shows an example of typical embodiment of this invention, and, thereby, the range of this invention is not interpreted narrowly.
＜外皮系組織産生促進用組成物について＞
本発明に係る外皮系組織産生促進用組成物は、ツツジ科エリカ属植物由来成分を少なくとも含有する。ツツジ科エリカ属の株種は特に限定されない。例えば、Ｅｒｉｃａ ｍｕｌｔｉｆｌｏｒａ、Ｅｒｉｃａ ｖｅｎｔｒｉｃｏｓａ、Ｅｒｉｃａ ｓｐａｒｓａ、Ｅｒｉｃａ ａｂｉｅｔｉｎａ、Ｅｒｉｃａ ｖｅｒｓｉｃｏｌｏｒ、Ｅｒｉｃａ ｏａｔｅｓｉｉ、Ｅｒｉｃａ ｃａｒｎｅａ、Ｅｒｉｃａ ｃｉｎｅｒｅａ、Ｅｒｉｃａ ｃｉｌｉａｒｉｓ、Ｅｒｉｃａ ｇｒａｃｉｌｉs、Ｅｒｉｃａ ｃｏｎｉｃａ、Ｅｒｉｃａ ｃｏｌｏｒａｎｓ、Ｅｒｉｃａ ｄａｒｌｅｙｅｎｓｉｓ、Ｅｒｉｃａ ｂａｕｅｒａ、Ｅｒｉｃａ ｖａｇａｎｓ、Ｅｒｉｃａ ｐａｔｅｒｓｏｎｉａ、Ｅｒｉｃａ ｆａｓｔｉｇｉａｔａ、Ｅｒｉｃａ ｐｌｕｋｅｎｅｔｉｉ、Ｅｒｉｃａ ｖｅｓｔｉｔａ、Ｅｒｉｃａ ｐｅｚｉｚａ、Ｅｒｉｃａ ｐｅｒｓｐｉｃｕａ（Ｅｒｉｃａ ｌｉｎｎａｅｏｉｄｅｓ）、Ｅｒｉｃａ ｍｅｌａｎｔｈｅｒａ、Ｅｒｉｃａ ｍｏｎｓｏｎｉａｎａ、Ｅｒｉｃａ ｒｅｇｉａ、Ｅｒｉｃａ ｗａｌｌｋｅｒｉａ、Ｅｒｉｃａ ｂｌａｎｄｆｏｒｄｉａ、Ｅｒｉｃａ ｃａｎａｌｉｃｕｌａｔａ、Ｅｒｉｃａ ｆｏｒｍｏｓａ、Ｅｒｉｃａ ｃｅｒｉｎｔｈｏｉｄｅｓなどを挙げることができる。
<About the composition for promoting the production of outer skin tissue>
The composition for promoting skin tissue production according to the present invention contains at least a component derived from the genus Ericaceae. The strain of the genus Ericaceae is not particularly limited. For example, Erica multiflora, Erica ventricosa, Erica sparsa, Erica abietina, Erica versicolor, Erica oatesii, Erica carnea, Erica cinerea, Erica ciliaris, Erica gracilis, Erica conica, Erica colorans, Erica darleyensis, Erica bauera, Erica vagans, Erica patersonia, Erica fastigita, Erica plukenetii, Erica vestita, Erica peziza, Erica perspica (Erica linnaeoides), Erica mela thera, Erica monsoniana, Erica regia, Erica wallkeria, Erica blandfordia, Erica canaliculata, Erica formosa, and the like Erica cerinthoides.
前記ツツジ科エリカ属植物は、葉、花、茎、根のいずれの部分も用いることができる。これらは、生の状態で用いてもよく、あるいはそれらの乾燥物を用いてもよい。更には、それらの抽出物を用いてもよい。抽出物を用いる場合、その抽出方法は特に限定されないが、一例としては、各種溶媒を用いた抽出法、熱水抽出法、酵素処理法、超臨界抽出法が挙げられる。抽出溶媒としては、メタノール、エタノール、プロパノール、ブタノール、プロピレングリコール、１，３−ブチレングリコール、グリセリン、アセトン、メチルエチルケトン、酢酸エチル、エーテル類、クロロホルム、及びジクロロメタンのような有機溶媒ならびに水を用いることができる。このうち、本発明においては、メタノール、エタノール、酢酸エチルまたはこれら溶媒と水との組合せが好ましく、毒性が低いという生体内における安全性を考慮すれば、エタノール、または水とエタノールとの混合溶媒がさらに好ましい。好適な抽出方法としては、乾燥したツツジ科エリカ属植物を、７０％エタノール中に１週間程度浸漬した後、孔径０．４５μｍのマイクロフィルターなどを用いて不溶物を除去することにより得ることができる。 Any of the leaves, flowers, stems, and roots can be used for the azalea family Erika. These may be used in the raw state, or their dried products may be used. Furthermore, those extracts may be used. In the case of using an extract, the extraction method is not particularly limited, and examples include extraction methods using various solvents, hot water extraction methods, enzyme treatment methods, and supercritical extraction methods. As an extraction solvent, an organic solvent such as methanol, ethanol, propanol, butanol, propylene glycol, 1,3-butylene glycol, glycerin, acetone, methyl ethyl ketone, ethyl acetate, ethers, chloroform, and dichloromethane, and water can be used. it can. Among these, in the present invention, methanol, ethanol, ethyl acetate or a combination of these solvents and water is preferable, and in view of safety in vivo that toxicity is low, ethanol or a mixed solvent of water and ethanol is used. Further preferred. As a suitable extraction method, it can be obtained by immersing a dried ericaceae plant in 70% ethanol for about one week and then removing insolubles using a microfilter having a pore size of 0.45 μm. .
以下、本発明に係る外皮系組織産生促進用組成物の主な作用を説明する。 Hereinafter, the main effects of the composition for promoting the production of the outer skin tissue according to the present invention will be described.
（１）発毛促進作用
本発明に係る外皮系組織産生促進用組成物は、発毛促進作用を有する。本発明に係る外皮系組織産生促進用組成物の発毛促進作用は、毛の成長を促せば、毛の状態は特に限定されず、毛周期における休止期の状態、即ち、毛が生えていない状態、及び既に毛が生えている状態のいずれにも効果を有する。
(1) Hair growth promoting action The composition for promoting the production of integumental tissue according to the present invention has a hair growth promoting action. The hair growth promoting action of the composition for promoting the production of integumental tissue according to the present invention is not particularly limited as long as hair growth is promoted, and the resting state in the hair cycle, that is, the hair is not growing It has an effect on both the state and the state where hair has already grown.
（２）毛乳頭細胞増殖促進作用
本発明に係る外皮系組織産生促進用組成物は、毛乳頭細胞増殖促進作用を有する。毛乳頭細胞は、毛包構成細胞に栄養および各種シグナル分子を供給し、毛母の機能調節に重要な役割を果たしている。毛乳頭細胞増殖が促進されることにより、ヘアマトリックス（毛母基）における毛包構成角化細胞の増殖も促進され、発毛が促される。このように、本発明に係る外皮系組織産生促進用組成物は、毛乳頭細胞増殖を促進させることにより、発毛効果を発揮している。
(2) Hair papillary cell proliferation promoting action The composition for promoting the production of outer skin tissue according to the present invention has a hair papillary cell growth promoting action. Papilla cells supply nutrients and various signal molecules to the follicle-constituting cells and play an important role in regulating the function of the hair matrix. By promoting the growth of dermal papilla cells, the growth of hair follicle keratinocytes in the hair matrix (hair matrix) is also promoted, and hair growth is promoted. Thus, the composition for promoting the production of integumental tissue according to the present invention exhibits a hair growth effect by promoting the proliferation of dermal papilla cells.
本発明に係る外皮系組織産生促進用組成物の毛乳頭細胞増殖促進作用のメカニズムの一例としては、毛乳頭細胞の細胞周期を促進させることにより、毛乳頭細胞増殖促進作用を有すると考えられる。細胞周期について、図１を用いて説明する。 As an example of the mechanism of the dermal papilla cell proliferation promoting action of the composition for promoting the production of integumental tissue according to the present invention, it is considered that the dermal papilla cell proliferation promoting action is promoted by promoting the cell cycle of the dermal papilla cells. The cell cycle will be described with reference to FIG.
図１は、一般的な細胞周期を示す概念図である。細胞周期とは、ある細胞分裂から次の細胞分裂までの期間をいう。大多数の細胞は、細胞周期から外れた静止期（Ｇ０期）の状態にあり、増殖刺激が加わると第一間期（Ｇ１期）、次いでＤＮＡ合成期（Ｓ期）に進み、ＤＮＡ合成を行う。ＤＮＡ合成が完了すると第二間期（Ｇ２期）に進み、細胞分裂の準備が行われる。該準備が整い、ゲノム複製が完了すると分裂期（Ｍ期）に進み、細胞分裂が開始される。そして、同一の遺伝情報を持つ２個の細胞に増え、再び第一間期（Ｇ１期）に戻る。細胞への増殖刺激が続く場合には、更にＤＮＡ合成期（Ｓ期）に進み、細胞周期を繰り返す。細胞刺激を受けなければ、静止期（Ｇ０期）に留まる。 FIG. 1 is a conceptual diagram showing a general cell cycle. The cell cycle refers to the period from one cell division to the next. The majority of cells are in a stationary phase (G0 phase) out of the cell cycle. When proliferation stimulation is applied, they proceed to the first interphase (G1 phase) and then to the DNA synthesis phase (S phase). Do. When DNA synthesis is completed, the process proceeds to the second interphase (G2 phase) and preparation for cell division is performed. When the preparation is complete and genome replication is completed, the cell proceeds to the division phase (M phase) and cell division is started. And it increases to two cells with the same genetic information, and returns to the first interphase (G1 phase) again. When cell growth stimulation continues, the cell cycle further repeats the DNA synthesis phase (S phase). If the cell is not stimulated, it remains in the stationary phase (G0 phase).
本発明に係る外皮系組織産生促進用組成物は、前記の毛乳頭細胞周期を促進させる。すると、毛乳頭細胞の分裂が進み、毛乳頭細胞が増殖する。そして、毛乳頭細胞から毛包構成細胞へ栄養および各種シグナル分子が十分に供給され、ヘアマトリックス（毛母基）における毛包構成角化細胞の分裂も促進される。このようにして、本発明に係る外皮系組織産生促進用組成物は、発毛を促進する。 The composition for promoting the production of the integumental tissue according to the present invention promotes the dermal papilla cell cycle. Then, the dermal papilla cells continue to divide and the dermal papilla cells proliferate. Nutrition and various signal molecules are sufficiently supplied from the hair papilla cells to the hair follicle constituent cells, and the division of the hair follicle constituent keratinocytes in the hair matrix (hair matrix) is also promoted. In this way, the composition for promoting the production of the outer skin tissue according to the present invention promotes hair growth.
（３）毛周期成長期誘導作用
本発明に係る外皮系組織産生促進用組成物は、毛周期成長期誘導作用を有する。毛周期について、図２を用いて説明する。
(3) Hair cycle growth phase inducing action The composition for promoting the production of integumental tissue according to the present invention has a hair cycle growth phase inducing action. The hair cycle will be described with reference to FIG.
図２は、皮膚の断面を示す模式図である。図２中の符号１〜８は、それぞれ、毛乳頭１、ヘアマトリックス（毛母基）２、外毛根鞘３、内毛根鞘４、立毛筋５、皮脂腺６、棍毛７、毛包バルジ領域８、新生（中）毛９を示している。毛周期は、成長期（図２中（Ｉ）参照）、退行期（図２中（II）参照）、休止期（図２中（III）参照）の３期に分かれている。 FIG. 2 is a schematic view showing a cross section of the skin. Reference numerals 1 to 8 in FIG. 2 denote a hair papilla 1, a hair matrix (matrix base) 2, an outer root sheath 3, an inner root sheath 4, napped muscle 5, a sebaceous gland 6, an eyelash 7, and a hair follicle bulge region. 8 shows new (middle) hair 9. The hair cycle is divided into three phases: a growth phase (see (I) in FIG. 2), a regression phase (see (II) in FIG. 2), and a resting phase (see (III) in FIG. 2).
成長期（Ｉ）では、毛乳頭１からヘアマトリックス（毛母基）２の毛包構成細胞へ栄養および各種シグナル分子が供給され、毛包構成角化細胞が分裂する。２個に分裂した毛包構成角化細胞のうち１個は、ヘアマトリックス（毛母基）２に留まって次の分裂に備え、他の１個は脱核、角化し、毛軸となり皮膚上へ伸長する。このようなヘアマトリックス（毛母基）２における毛包構成細胞の分裂が次々に起こり、毛が成長する。 In the growth period (I), nutrients and various signal molecules are supplied from the hair papilla 1 to the hair follicle constituent cells of the hair matrix (hair matrix) 2, and the hair follicle constituent keratinocytes divide. One of the follicle-constituting keratinocytes divided into two stays in the hair matrix 2 to prepare for the next division, and the other one is denucleated and keratinized to become the hair shaft on the skin Elongate. The hair follicle constituent cells in the hair matrix (hair matrix) 2 divide one after another, and hair grows.
一定期間、成長期（Ｉ）を経ると、退行期（II）に入る。退行期（II）では、毛乳頭１が萎縮していき、それに伴って、前記毛包構成角化細胞の分裂が止まり、毛包が皮膚表面へと退行し、毛の成長も止まる。 After a certain period of growth (I), it enters a regression (II). In the regression phase (II), the hair papilla 1 shrinks, and accordingly, the division of the keratinocytes of the hair follicle stops, the hair follicle retreats to the skin surface, and hair growth also stops.
休止期（III）では、毛は、下端部がやや膨らんだ棍毛７となり、棍毛７の下端部が、立毛筋５の付着する毛包バルジ領域８まで上昇した状態で留まるか、若しくは脱落する。 In the rest period (III), the hairs become the eyelashes 7 whose lower end portion is slightly swollen, and the lower end portion of the eyelashes 7 remains in a state where it rises up to the hair follicle bulge region 8 to which the raised hair 5 adheres or falls off. To do.
やがて、残存する外毛根鞘３の下端部に、新たに毛乳頭１とヘアマトリックス（毛母基）２が形成され、再びヘアマトリックス（毛母基）２における毛包構成角化細胞の分裂が開始し、成長期（Ｉ）へと入る。 Eventually, the hair papilla 1 and the hair matrix (matrix group) 2 are newly formed at the lower end of the remaining outer root sheath 3, and the division of the hair follicle keratinocytes in the hair matrix (matrix group) 2 again occurs. Start and enter the growth period (I).
本発明に係る外皮系組織産生促進用組成物は、特に休止期（III）の状態から成長期（Ｉ）へと誘導する。その結果、発毛が促進される。 The composition for promoting the production of integumental tissue according to the present invention is particularly induced from the resting state (III) to the growth phase (I). As a result, hair growth is promoted.
（４）血管拡張作用
本発明に係る外皮系組織産生促進用組成物は、血管拡張作用を有する。血管拡張作用を有することにより、血流が促進され、外皮系組織細胞の増殖を促し、外皮系組織産生が促進される。一例を挙げると、本発明の血管拡張作用により、外皮系組織の血流が促進され、毛包構成細胞等の増殖を促し、発毛等が促進される。
(4) Vasodilatory action The composition for promoting the production of integumental tissue according to the present invention has a vasodilatory action. By having a vasodilatory effect, blood flow is promoted, the proliferation of integumentary tissue cells is promoted, and integumentary tissue production is promoted. For example, the blood vessel expansion action of the present invention promotes blood flow in the integumental tissue, promotes proliferation of hair follicle constituent cells, and promotes hair growth.
（５）メラニン産生促進作用
本発明に係る外皮系組織産生促進用組成物は、メラニン産生促進作用を有する。本発明に係る外皮系組織産生促進用組成物が作用するメラニンは、その存在組織は特に限定されず、皮膚、毛髪、眼球など色素細胞（メラノサイト）を有する組織に存在するメラニン全てに作用する。例えば、皮膚のメラニンを増殖させることにより、尋常性白皮症等の色素欠乏症、白皮症などの予防や治療に用いることが可能である。また、例えば、毛髪のメラニンを増殖させることにより、白髪症の予防や治療に用いることも可能である。更に、メラニン産生促進作用と、前記（１）から（４）の作用とが同時に作用することにより、黒い毛を発毛させることも可能である。
(5) Melanin production promoting action The composition for promoting skin tissue production according to the present invention has a melanin production promoting action. Melanin to which the composition for promoting the production of integumental tissue according to the present invention acts is not particularly limited, and acts on all melanin present in tissues having pigment cells (melanocytes) such as skin, hair, and eyeball. For example, by proliferating melanin in the skin, it can be used for the prevention and treatment of pigment deficiency such as vulgaris vulgaris and albinism. In addition, for example, it can be used for the prevention or treatment of gray hair by proliferating melanin in hair. Further, the melanin production promoting action and the actions (1) to (4) act simultaneously, whereby black hair can be made to grow.
（６）チロシナーゼ合成促進作用
本発明に係る外皮系組織産生促進用組成物は、チロシナーゼ合成促進作用を有する。チロシナーゼを合成する組織は特に限定されず、皮膚、毛髪、眼球など色素細胞（メラノサイト）を有し、チロシナーゼの合成が行われる組織全てにおいて、チロシナーゼ合成を促進する。本発明に係る外皮系組織産生促進用組成物は、チロシナーゼ合成促進作用を有するので、例えば、先天的にチロシナーゼが欠乏する先天性白皮症（アルビノ）等の治療に用いることが可能である。また、尋常性白皮症等の色素欠乏症、白皮症、白髪症などの予防や治療に用いることも可能である。
(6) Tyrosinase synthesis promoting action The composition for promoting skin tissue production according to the present invention has a tyrosinase synthesis promoting action. The tissue that synthesizes tyrosinase is not particularly limited, and tyrosinase synthesis is promoted in all tissues that have pigment cells (melanocytes) such as skin, hair, and eyeball and that synthesize tyrosinase. Since the composition for promoting the production of integumental tissue according to the present invention has a tyrosinase synthesis promoting action, it can be used, for example, for the treatment of congenital albinism (albino) or the like, which is inherently deficient in tyrosinase. It can also be used for the prevention and treatment of pigment deficiencies such as acne vulgaris, albinism, and white hair.
＜医薬品について＞
本発明に係る外皮系組織産生促進用組成物は、これを有効成分として、医薬品に適用することができる。本発明に係る医薬品は、本発明に係る外皮系組織産生促進用組成物を含むもの全てを含有する。
<About drugs>
The composition for promoting the production of the integumental tissue according to the present invention can be applied to pharmaceuticals using this as an active ingredient. The pharmaceutical products according to the present invention include all those containing the composition for promoting the production of integumental tissues according to the present invention.
本発明に係る医薬品は、単独で用いることもでき、また既存のあらゆる薬剤等と併用することができる。更には、本発明の発毛効果を損なわない範囲において、既存のあらゆる薬剤等と合剤とすることもできる。 The pharmaceutical agent according to the present invention can be used alone or in combination with all existing drugs and the like. Furthermore, in the range which does not impair the hair growth effect of this invention, it can also be set as a mixture with all the existing chemical | medical agents.
本発明に係る医薬品の剤型は特に限定されない。一例としては、外用液剤、外用ゲル剤、クリーム剤、軟膏剤、スプレー剤、点鼻液剤等の外用剤、散剤、細粒剤、顆粒剤、錠剤、カプセル剤、懸濁液、エマルジョン剤、シロップ剤、エキス剤、丸剤等の経口剤、又は注射剤として用いることができる。 The pharmaceutical dosage form according to the present invention is not particularly limited. Examples include external preparations, external gels, creams, ointments, sprays, nasal drops, etc., powders, fine granules, granules, tablets, capsules, suspensions, emulsions, syrups It can be used as an oral preparation such as an agent, an extract and a pill, or as an injection.
前記外用剤には、有効成分である本発明に係る外皮系組織産生促進用組成物に加え、例えば、基材、保存剤、乳化剤、着色剤、防腐剤、界面活性剤、酸化防止剤等の、医薬製剤の分野で通常使用し得る全ての添加剤を含有させることができる。 In addition to the composition for promoting the production of the outer skin tissue according to the present invention, which is an active ingredient, the external preparation includes, for example, a base material, a preservative, an emulsifier, a colorant, a preservative, a surfactant, an antioxidant, and the like. In addition, all additives which can be usually used in the field of pharmaceutical preparations can be contained.
前記経口剤には、有効成分である本発明に係る外皮系組織産生促進用組成物に加え、例えば、賦形剤、結合剤、崩壊剤、界面活性剤、保存剤、着色剤、矯味剤、香料、安定化剤、防腐剤、酸化防止剤等の、医薬製剤の分野で通常使用し得る全ての添加剤を含有させることができる。
また、ドラックデリバリーシステム（ＤＤＳ）を利用して、徐放性製剤等にすることもできる。
In addition to the composition for promoting the production of the integumental tissue according to the present invention, which is an active ingredient, for example, excipients, binders, disintegrants, surfactants, preservatives, coloring agents, flavoring agents, All additives commonly used in the field of pharmaceutical preparations such as fragrances, stabilizers, preservatives and antioxidants can be contained.
In addition, a sustained-release preparation can be obtained using a drug delivery system (DDS).
前記注射剤には、有効成分である本発明に係る外皮系組織産生促進用組成物に加え、例えば、溶剤、安定剤、溶解補助剤、懸濁化剤、保存剤、等張化剤、防腐剤、酸化防止剤等の、医薬製剤の分野で通常使用し得る全ての添加剤を含有させることができる。 In addition to the composition for promoting the production of the integumental tissue according to the present invention, which is an active ingredient, the injection includes, for example, a solvent, a stabilizer, a solubilizer, a suspending agent, a preservative, an isotonic agent, an antiseptic agent, and the like. All additives that can be usually used in the field of pharmaceutical preparations, such as agents and antioxidants, can be contained.
本発明に係る医薬品は、その有効成分が天然由来成分であるため、多剤との併用を注意する必要性が低い。そのため、種々の疾患を罹患した患者に対しても安心して投与できる可能性も高い。また、長期間、連続的に投与しても副作用を心配する必要性も少ない。 Since the active ingredient is a naturally-derived ingredient, the pharmaceutical product according to the present invention has a low need for attention in combination with multiple drugs. Therefore, there is a high possibility that it can be administered safely to patients suffering from various diseases. Moreover, there is little need to worry about side effects even if administered continuously for a long period of time.
＜飲食物について＞
本発明に係る外皮系組織産生促進用組成物は、これを有効成分として、飲食物に適用することができる。本発明に係る飲食物は、本発明に係る外皮系組織産生促進用組成物を含むもの全てを含有する。
<About food and drink>
The composition for promoting the production of integumental tissue according to the present invention can be applied to food and drink as an active ingredient. The food and drink according to the present invention includes all of the foods including the composition for promoting the production of the outer skin tissue according to the present invention.
例えば、本発明に係る外皮系組織産生促進用組成物は、保健機能食品（特定保健機能食品、栄養機能食品、飲料を含む）や、いわゆる健康食品（飲料を含む）、又は飼料に含有させることができる。また、口中に一時的に含むもの、例えば、歯磨剤、染口剤、チューインガム等に含有させることもできる。 For example, the composition for promoting the production of integumental tissue according to the present invention is contained in health functional foods (including specific health functional foods, nutritional functional foods and beverages), so-called health foods (including beverages), or feed. Can do. Moreover, it can also be contained in what is temporarily contained in a mouth, for example, a dentifrice, a mouth dye, a chewing gum etc.
前記飲食物には、有効成分である本発明に係る外皮系組織産生促進用組成物に加え、例えば、保存剤、着色剤、防腐剤、酸化防止剤等の、食品分野で通常使用し得る全ての添加剤を含有させることができる。また、他の有効成分を、必要に応じて適宜含有させることもできる。 In the food and drink, in addition to the composition for promoting the production of the outer skin tissue according to the present invention, which is an active ingredient, for example, all of the foods such as preservatives, coloring agents, preservatives, antioxidants and the like that can be normally used Additives can be included. Further, other active ingredients can be appropriately contained as necessary.
本発明に係る外皮系組織産生促進用組成物は、天然由来成分であるため、飲食物との相互作用を注意する必要性が低く、あらゆる飲食物に含有させることができる。また、安全性が高いため、長期間、連続的に摂取が可能である。 Since the composition for promoting the production of outer skin tissue according to the present invention is a naturally derived component, it is less necessary to pay attention to the interaction with food and drink, and can be contained in any food and drink. Moreover, since it is highly safe, it can be taken continuously for a long period of time.
＜化粧品について＞
本発明に係る外皮系組織産生促進用組成物は、これを有効成分として、化粧品に適用することができる。本発明に係る化粧品は、本発明に係る外皮系組織産生促進用組成物を含むもの全てを含有する。
<About cosmetics>
The composition for promoting skin tissue production according to the present invention can be applied to cosmetics using this as an active ingredient. The cosmetics according to the present invention include all those containing the composition for promoting the production of integumental tissues according to the present invention.
例えば、本発明に係る外皮系組織産生促進用組成物は、ローション、乳液、クリーム、ジェル、美溶液、エアゾール等のあらゆる化粧品に含有させることができる。 For example, the composition for promoting skin tissue production according to the present invention can be contained in all cosmetics such as lotions, emulsions, creams, gels, beauty solutions, and aerosols.
前記化粧品には、有効成分である本発明に係る外皮系組織産生促進用組成物に加え、例えば、基材、保存剤、乳化剤、着色剤、防腐剤、界面活性剤、紫外線吸収剤、酸化防止剤等の、化粧品分野で通常使用し得る全ての添加剤を含有させることができる。また、他の有効成分を、必要に応じて適宜含有させることもできる。 For the cosmetics, in addition to the composition for promoting the production of the outer skin tissue according to the present invention which is an active ingredient, for example, a base material, a preservative, an emulsifier, a coloring agent, a preservative, a surfactant, an ultraviolet absorber, an antioxidant All additives that can be used normally in the cosmetic field, such as agents, can be included. Further, other active ingredients can be appropriately contained as necessary.
本発明に係る外皮系組織産生促進用組成物は、天然由来成分であるため、他の成分との相互作用を注意する必要性が低く、あらゆる化粧品に含有させることができる。また、安全性が高いため、長期間、連続的な使用が可能である。 Since the composition for promoting the production of integumental tissue according to the present invention is a naturally derived component, it is less necessary to pay attention to the interaction with other components, and can be contained in any cosmetic. Moreover, since it is highly safe, it can be used continuously for a long time.
実施例１では、実施例２から実施例４で使用するツツジ科エリカ属植物由来成分を含有する外皮系組織産生促進用組成物を作製した。本実施例では、前記外皮系組織産生促進用組成物として、ツツジ科エリカ属のＥｒｉｃａ ｍｕｌｔｉｆｌｏｒａから抽出した抽出物（以下「エリカ抽出物」と称する。）を用いた。 In Example 1, a composition for promoting the production of integumental tissues containing components derived from the genus Ericaceae used in Examples 2 to 4 was prepared. In this example, an extract extracted from Erica multiflora belonging to the genus Ericaceae (hereinafter referred to as “Erica extract”) was used as the composition for promoting the production of integumental tissues.
具体的には、乾燥したツツジ科エリカ属のＥｒｉｃａ ｍｕｌｔｉｆｌｏｒａ１００ｇを、７０％エタノール１０００ｍｌで１週間浸漬した後、孔径０．４５μｍのマイクロフィルターを用いて濾過、精製し、以下の実施例２から実施例１３で用いるエリカ抽出物を得た。 Specifically, after 100 g of dried Erica multiflora belonging to the genus Ericaceae is immersed in 1000 ml of 70% ethanol for 1 week, it is filtered and purified using a microfilter having a pore diameter of 0.45 μm. The Erika extract used in 13 was obtained.
このエリカ抽出物をロータリーエバポレーターにより５０℃で減圧乾燥を行い、やや緑色がかった薄い褐色の粉末（エリカ抽出乾燥物）を得た。その収量は１．２％であった。 This Erica extract was dried under reduced pressure at 50 ° C. with a rotary evaporator to obtain a slightly brownish-red powder (Erica extract dried product) with a slight green color. The yield was 1.2%.
実施例２、及び実施例３では、本発明に係る外皮系組織産生促進用組成物の毛乳頭細胞増殖促進作用について調べた。 In Example 2 and Example 3, the dermal papilla cell proliferation promoting action of the composition for promoting the production of the outer skin tissue according to the present invention was examined.
まず、実施例２では、本発明に係る外皮系組織産生促進用組成物をヒト毛乳頭細胞へ添加した場合の実際の毛乳頭細胞増殖促進作用をＭＴＴアッセイにより検討した。 First, in Example 2, the actual dermal papilla cell proliferation promoting action when the composition for promoting the production of the integumental tissue according to the present invention was added to human dermal papilla cells was examined by MTT assay.
ＭＴＴアッセイは、相対的な生存細胞数を測定する方法である。ＭＴＴ（３−（４，５−ｄｉｍｅｔｈｙｌｔｈｉａｚｏｌ−２−ｙｌ）−２，５−ｄｉｐｈｅｎｙｌｔｅｔｒａｚｏｌｉｕｍ ｂｒｏｍｉｄｅ）は、生細胞中のミトコンドリア内脱水素酵素（フタル酸脱水素酵素）により、ＭＴＴホルマザンに還元される。そのため、生産されたＭＴＴホルマザン量と生存細胞数は比例する。従って、培養細胞中にＭＴＴ試薬を添加した後、５７０ｎｍにおける吸光度を測定してＭＴＴホルマザン量（相対値）を取得することにより、相対的な生存細胞数を測定できる。 The MTT assay is a method for measuring the relative number of viable cells. MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) is reduced to MTT formazan by mitochondrial dehydrogenase (phthalate dehydrogenase) in living cells. Therefore, the amount of MTT formazan produced is proportional to the number of viable cells. Therefore, the relative number of viable cells can be determined by adding the MTT reagent to the cultured cells and measuring the absorbance at 570 nm to obtain the amount of MTT formazan (relative value).
そこで、本実施例では、ヒト毛乳頭細胞の培養細胞に、実施例１で抽出したエリカ抽出物を添加した後、ＭＴＴアッセイを行い、ヒト毛乳頭細胞の生存細胞数を測定することにより、エリカ抽出物の毛乳頭細胞増殖促進作用の有無を調べた。 Therefore, in this example, the Erika extract extracted in Example 1 was added to the cultured cells of human hair papilla cells, and then MTT assay was performed to measure the number of viable cells of human hair papilla cells. The extract was examined for the effect of promoting the proliferation of dermal papilla cells.
具体的には、まず、ヒト毛乳頭細胞に、実施例１で抽出したエリカ抽出物を添加した。添加するエリカ抽出物の希釈倍率は、１．０×１０-６、１．０×１０-５、１．０×１０-４、１．０×１０-３、１．０×１０-２とした。そして、ＭＴＴ法に準拠して５７０ｎｍの吸光度を測定し、ＭＴＴホルマザン量（相対値）を取得した。 Specifically, first, the Erika extract extracted in Example 1 was added to human hair papilla cells. The dilution ratio of the Erika extract to be added is 1.0 × 10 −6 , 1.0 × 10 −5 , 1.0 × 10 −4 , 1.0 × 10 −3 , and 1.0 × 10 −2 . did. And the light absorbency of 570 nm was measured based on MTT method, and the amount (relative value) of MTT formazan was acquired.
結果を図３に示す。図３に示す通り、本発明に係る外皮系組織産生促進用組成物であるエリカ抽出物の添加により、ヒト毛乳頭細胞のＭＴＴホルマザン量が増加していることが確認できる。特に、添加倍率１．０×１０-３を超えると、急激に増加している。 The results are shown in FIG. As shown in FIG. 3, it can be confirmed that the amount of MTT formazan in human hair papilla cells is increased by the addition of Erika extract, which is a composition for promoting the production of integumental tissue according to the present invention. In particular, when the addition ratio exceeds 1.0 × 10 −3 , it rapidly increases.
以上から、ヒト毛乳頭細胞にエリカ抽出物を添加することにより、ヒト毛乳頭細胞の生存細胞数が増加することが分かった。従って、本発明に係る外皮系組織産生促進用組成物（エリカ抽出物）は、毛乳頭細胞増殖促進作用を有することが示された。 From the above, it was found that the number of viable cells of human hair papilla cells increased by adding Erika extract to human hair papilla cells. Therefore, it was shown that the composition for promoting the production of integumental tissue (Erica extract) according to the present invention has an action to promote proliferation of hair papilla cells.
次に、実施例３では、本発明に係る外皮系組織産生促進用組成物をヒト毛乳頭細胞添加することにより、毛乳頭細胞の細胞周期促進活性について調べた。 Next, in Example 3, the cell cycle promoting activity of dermal papilla cells was examined by adding the human dermal papilla cells to the composition for promoting the production of integumental tissues according to the present invention.
ヒト毛乳頭細胞の核ＤＮＡを蛍光ラベリングし、フローサイトメーターを用いて、細胞周期を同定した。各細胞の大きさとＤＮＡ含有量を調べることにより、細胞周期のステージを推測することができる。例えば、Ｇ０・Ｇ１期のＤＮＡ含有量を１倍量とすると、Ｓ期は１〜２倍量、Ｇ２・Ｍ期は２〜１倍量となる。結果を図４、及び図５に示す。 The nuclear DNA of human hair papilla cells was fluorescently labeled and the cell cycle was identified using a flow cytometer. By examining the size and DNA content of each cell, the stage of the cell cycle can be estimated. For example, assuming that the DNA content in the G0 / G1 period is 1 time, the S period is 1 to 2 times, and the G2 · M period is 2 to 1 times. The results are shown in FIG. 4 and FIG.
次に、実施例１で抽出したエリカ抽出物を前記ヒト毛乳頭細胞へ添加した。添加から１２時間後に、前記と同様に、ヒト毛乳頭細胞の細胞周期を同定した。結果を図６、及び図７に示す。また、エリカ抽出物添加前と、添加後のそれぞれの細胞周期における細胞割合の比較を表１に示す。 Next, the Erika extract extracted in Example 1 was added to the human hair papilla cells. 12 hours after the addition, the cell cycle of human hair papilla cells was identified as described above. The results are shown in FIG. 6 and FIG. In addition, Table 1 shows a comparison of the cell ratio in each cell cycle before and after the addition of the Erika extract.
表１に示す通り、エリカ抽出物の添加前と添加後のＧ２・Ｍ期の細胞が、８．２％から１０．０％へと増加していることが分かる。これは細胞周期及び細胞分裂が促進されたことを示す。 As shown in Table 1, it can be seen that the G2 · M phase cells before and after the addition of Erika extract increased from 8.2% to 10.0%. This indicates that cell cycle and cell division were promoted.
実施例３の結果から、本発明に係る外皮系組織産生促進用組成物は、毛乳頭細胞の細胞周期促進、及び細胞分裂促進により、毛乳頭細胞増殖促進作用を有することが分かった。 From the results of Example 3, it was found that the composition for promoting the production of the integumental tissue according to the present invention has the effect of promoting the proliferation of dermal papilla cells by promoting the cell cycle and promoting cell division of dermal papilla cells.
実施例４では、本発明に係る外皮系組織産生促進用組成物の毛周期成長期誘導作用、及び血管拡張作用について、Ｃ３Ｈ／ＨｅＮマウスを用いて調べた。８週齢のＣ３Ｈ／ＨｅＮマウスは、毛周期がちょうど２周期目で休止期にて同調しているため、発毛試験などに多く用いられる。 In Example 4, the hair cycle growth phase-inducing action and vasodilatory action of the composition for promoting the production of integumental tissue according to the present invention were examined using C3H / HeN mice. 8-week-old C3H / HeN mice are frequently used for hair growth tests and the like because the hair cycle is exactly the second cycle and synchronized in the resting phase.
８週齢の雄性Ｃ３Ｈ／ＨｅＮマウスの体毛をバリカンにて剃った後、表２に示す試験群に分けて皮膚色の変化を３〜４週間後に確認した。また皮膚色の変化を確認後、血管サイズを観察した。 After shaving the hair of 8-week-old male C3H / HeN mice with clippers, they were divided into the test groups shown in Table 2 and changes in skin color were confirmed after 3-4 weeks. After confirming the change in skin color, the blood vessel size was observed.
コントロール群の結果を図８に、Ａ群の結果を図９に、Ｂ群の結果を図１０に、Ｃ群の結果を図１１に示す。皮膚色が黒化した部分が毛周期中の成長期を示し、白い部分が毛周期中の休止期を示す。 FIG. 8 shows the results of the control group, FIG. 9 shows the results of the group A, FIG. 10 shows the results of the group B, and FIG. 11 shows the results of the group C. The part where the skin color is black indicates the growth period during the hair cycle, and the white part indicates the rest period during the hair cycle.
図８に示す通り、コントロール群においては、成長期の毛包の出現は見られるが、血管サイズは小さいままであった。 As shown in FIG. 8, in the control group, the appearance of hair follicles during the growth period was observed, but the blood vessel size remained small.
図９に示す通り、Ａ群においては、成長期の毛包が出現し、血管拡張も確認できた（図９中矢印Ｘ）。また、マウスによっては、次の休止期が観察できるものもあった（図９中矢印Ｙ）。次の休止期が出現したのは、成長期へと誘導された毛周期が正常に進んだことを意味すると考えられる。 As shown in FIG. 9, in group A, hair follicles in the growth phase appeared and vasodilation was also confirmed (arrow X in FIG. 9). Some mice were able to observe the next rest period (arrow Y in FIG. 9). The appearance of the next quiescent period is considered to mean that the hair cycle induced to the growth period has progressed normally.
図１０に示す通り、Ｂ群においては、成長期の毛包が出現し、血管拡張も確認できた（図１０中矢印Ｘ）。 As shown in FIG. 10, in the group B, hair follicles in the growth period appeared, and vasodilation was also confirmed (arrow X in FIG. 10).
図１１に示す通り、Ｃ群においては、成長期の毛包が出現し、血管拡張も確認できた（図１１中矢印Ｘ）。また、マウスによっては、次の休止期が観察できるものもあった（図１１中矢印Ｙ）。次の休止期が出現したのは、成長期へと誘導された毛周期が正常に進んだことを意味すると考えられる。 As shown in FIG. 11, in the group C, hair follicles in the growth phase appeared and vasodilation was also confirmed (arrow X in FIG. 11). Some mice were able to observe the next rest period (arrow Y in FIG. 11). The appearance of the next quiescent period is considered to mean that the hair cycle induced to the growth period has progressed normally.
実施例４の結果から、本発明に係る外皮系組織産生促進用組成物は、毛周期成長期誘導作用、及び血管拡張作用を有することが分かった。 From the results of Example 4, it was found that the composition for promoting the production of integumental tissue according to the present invention has a hair cycle growth phase inducing action and a vasodilating action.
実施例２から実施例４の結果を総合すると、本発明に係る外皮系組織産生促進用組成物の発毛促進作用は、毛乳頭細胞増殖が促進され、毛乳頭細胞から血管拡張促進又は血管内皮細胞増殖促進に関わる因子が分泌され血流が促進し、この血流増加により、栄養因子等が毛包へ補充され、その結果、発毛効果が得られるといった作用機序が示唆される。 Summarizing the results of Example 2 to Example 4, the hair growth promoting action of the composition for promoting the production of integumentary tissue according to the present invention promotes the proliferation of dermal papilla cells, promotes vasodilation from dermal papilla cells, or vascular endothelium. Factors related to cell growth promotion are secreted and blood flow is promoted, and this increase in blood flow suggests that a nutrient factor and the like are supplemented to the hair follicle, resulting in a hair growth effect.
実施例５では、本発明に係る外皮系組織産生促進用組成物のメラニン産生促進作用について検討した。具体的には、Ｂ１６細胞株を用いて行った。Ｂ１６細胞は、マウスのメラノーマ（黒色腫）から樹立された細胞株である。メラニン色素を産生する色素細胞であるため、メラニン合成に関する薬効試験に広く用いられている。 In Example 5, the melanin production promoting action of the composition for promoting skin tissue production according to the present invention was examined. Specifically, the B16 cell line was used. B16 cells are a cell line established from mouse melanoma (melanoma). Since it is a pigment cell that produces melanin pigment, it is widely used in medicinal effects tests on melanin synthesis.
１０％ウシ胎児血清含有のＤＭＥＭ培地１５ｍｌを含む細胞培養用１０ｃｍシャーレに、Ｂ１６細胞を、その細胞密度が１×１０６細胞/シャーレになるように接種し、５％ＣＯ2 下、３７℃にて一昼夜培養後、実施例１抽出したエリカ抽出物を、１/２００、１/２０００、１／２００００の濃度になるようにシャーレに添加し（投与群）、さらに同条件下で３日間培養を行った。コントロール群としては、溶媒のみを添加したものを用いた。 B16 cells are inoculated into a 10 cm petri dish for cell culture containing 15 ml of DMEM medium containing 10% fetal bovine serum so that the cell density becomes 1 × 10 6 cells / petre, and the temperature is 37 ° C. under 5% CO 2. After all day and night culture, the Erika extract extracted in Example 1 was added to the petri dish so as to have a concentration of 1/200, 1/2000, 1/20000 (administration group), and further cultured under the same conditions for 3 days. went. As a control group, a group to which only a solvent was added was used.
培養終了後、ＰＢＳ（−）緩衝液１０ｍｌで２回洗浄し、０．２５％トリプシン処理にて細胞を収集し、その後０．１％トライトンＸ１００含有ＰＢＳ（−）にて膜を溶解後、トリクロロ酢酸にてメラニンを抽出・精製、水酸化ナトリウムにメラニンを溶解した。そして、既存の方法により、メラニン含有量（μｇ）を測定した。結果を表３及び図１２に示す。 After completion of the culture, the cells were washed twice with 10 ml of PBS (−) buffer, cells were collected by 0.25% trypsin treatment, and then the membrane was lysed with PBS (−) containing 0.1% Triton X100. Melanin was extracted and purified with acetic acid, and melanin was dissolved in sodium hydroxide. And melanin content (microgram) was measured by the existing method. The results are shown in Table 3 and FIG.
表３及び図１２に示す通り、細胞内におけるメラニン産生量は、コントロール群では５．５３±０．７０ｐｇであったのに対して、１/２００投与群では４．７８±０．３８ｐｇ、１/２０００投与群では６．３０±０．３９ｐｇ、１/２００００投与群では７．３８±０．８０ｐｇ、と有意な上昇が確認された。 As shown in Table 3 and FIG. 12, the amount of melanin production in the cells was 5.53 ± 0.70 pg in the control group, whereas 4.78 ± 0.38 pg in the 1/200 administration group, A significant increase of 6.30 ± 0.39 pg in the / 2000 administration group and 7.38 ± 0.80 pg in the 1/20000 administration group was confirmed.
実施例５の結果から、本発明に係る外皮系組織産生促進用組成物が、色素細胞（メラノサイト）中のメラニン産生を促進することが分かった。 From the results of Example 5, it was found that the composition for promoting the production of integumental tissue according to the present invention promotes melanin production in pigment cells (melanocytes).
実施例６では、本発明に係る外皮系組織産生促進用組成物の細胞毒性を調べた。具体的には、実施例５と同様の投与群とコントロール群において、処置後４８時間の細胞生存率を、フローサイトメトリー法を用いて測定した。 In Example 6, the cytotoxicity of the composition for promoting the production of the outer skin tissue according to the present invention was examined. Specifically, in the same administration group and control group as in Example 5, the cell viability 48 hours after the treatment was measured using a flow cytometry method.
フローサイトメトリー法は、死細胞を染色する蛍光色素（ＰＭ２）と核内ＤＮＡを染色する蛍光色素（ＰＭ１）を用いて細胞を染色し、さらに前方散乱光の大きさにより凝集性細胞、アポトーシス細胞、ネクローシス細胞を区別して、生細胞と死細胞を判別する方法である。従って、ＰＭ２染色が低く、かつＰＭ１染色が強いものから、凝集細胞を除いたものをカウントすることにより、細胞生存率を測定することが可能である。 In the flow cytometry method, cells are stained using a fluorescent dye (PM2) that stains dead cells and a fluorescent dye (PM1) that stains nuclear DNA, and further, aggregated cells and apoptotic cells are determined by the magnitude of forward scattered light. This is a method for distinguishing between necrotic cells and distinguishing live cells from dead cells. Therefore, it is possible to measure the cell viability by counting the number of cells excluding aggregated cells from those having low PM2 staining and strong PM1 staining.
測定結果を表４及び図１３に示す。 The measurement results are shown in Table 4 and FIG.
表４及び図１３に示す通り、細胞生存率は、コントロール群では９８．４７±０．７０％であったのに対して、１/２００投与群では９７．７０±０．３８％、１/２０００投与群では９８．１７±０．３９％、１/２００００投与群では９８．１０±０．８０％、と同等であった。 As shown in Table 4 and FIG. 13, the cell survival rate was 98.47 ± 0.70% in the control group, compared with 97.70 ± 0.38% in the 1/200 administration group, The 2000 administration group was equivalent to 98.17 ± 0.39% and the 1/20000 administration group was 98.10 ± 0.80%.
実施例６の結果から、本発明に係る外皮系組織産生促進用組成物は細胞毒性を有さないことが分かった。 From the results of Example 6, it was found that the composition for promoting the production of integumental tissue according to the present invention has no cytotoxicity.
実施例７では、本発明に係る外皮系組織産生促進用組成物のチロシナーゼ合成促進作用について検討した。具体的には、ウェスタンブロッティング（Western blotting：ＷＢ）法を用いて行った。 In Example 7, the tyrosinase synthesis promoting action of the composition for promoting the production of the outer skin tissue according to the present invention was examined. Specifically, Western blotting (WB) method was used.
ウェスタンブロッティング (Western blotting; ＷＢ)法とは、電気泳動によって分離した蛋白質を膜に転写し、任意の蛋白質に対する抗体でその蛋白質の存在を検出する手法である。 The Western blotting (WB) method is a technique in which a protein separated by electrophoresis is transferred to a membrane, and the presence of the protein is detected with an antibody against an arbitrary protein.
まず、１０ｃｍディッシュにＢ１６細胞を３×１０６cells/ dishで播き、一晩培養後、実施例１抽出したエリカ抽出物を増殖培地で３００倍希釈し、細胞へ添加した（投与群）。そして、５時間、２４時間、４８時間培養後、細胞のタンパク質を抽出した。コントロール群としては、溶媒のみを添加したものを用いた。 First, B16 cells were seeded at 3 × 10 6 cells / dish in a 10 cm dish, and after overnight culture, the Erika extract extracted in Example 1 was diluted 300-fold with a growth medium and added to the cells (administration group). Then, after culturing for 5 hours, 24 hours, and 48 hours, cellular proteins were extracted. As a control group, a group to which only a solvent was added was used.
抽出したタンパク質をＳＤＳ−ＰＡＧＥ（１ウェルあたり１５μｇ、２００Ｖ、４５分）により分離し、ニトロセルロースメンブレンに転写した（１００Ｖ、０．３５Ａ、６０分）。一次抗体（Tyrosinase; Santa Cruz Biotechnology, Inc., USA, 200倍希釈）で２時間反応後、二次抗体（HRP-labeled anti-rabbit IgG; Santa Cruz Biotechnology, Inc., USA, 2000 倍希釈）で９０分反応させた。 The extracted protein was separated by SDS-PAGE (15 μg per well, 200 V, 45 minutes) and transferred to a nitrocellulose membrane (100 V, 0.35 A, 60 minutes). After reacting with primary antibody (Tyrosinase; Santa Cruz Biotechnology, Inc., USA, 200-fold dilution) for 2 hours, then with secondary antibody (HRP-labeled anti-rabbit IgG; Santa Cruz Biotechnology, Inc., USA, 2000-fold dilution) The reaction was performed for 90 minutes.
その後、発光試薬（Immobilon Western Chemiluminescent HRP Substrate: Millipore, USA）でタンパク質を検出し、写真を撮った。結果を図１４に示す。コントロール群をcontrolで、エリカ抽出物添加群をEricaで示す。 Thereafter, the protein was detected with a luminescent reagent (Immobilon Western Chemiluminescent HRP Substrate: Millipore, USA), and a photograph was taken. The results are shown in FIG. The control group is indicated by control, and the Erica extract added group is indicated by Erica.
図１４に示す通り、投与群では、５時間培養後から既にチロシナーゼの合成が確認できた。また、５時間、２４時間、４８時間培養後のいずれも、投与群の方がコントロール群に比べ、チロシナーゼの合成量が多いことが確認できた。 As shown in FIG. 14, in the administration group, the synthesis of tyrosinase was already confirmed after culturing for 5 hours. It was also confirmed that the amount of tyrosinase synthesized in the administration group was higher in the administration group than in the control group after culturing for 5 hours, 24 hours, and 48 hours.
実施例７の結果から、本発明に係る外皮系組織産生促進用組成物が、チロシナーゼ合成を促進することが分かった。 From the results of Example 7, it was found that the composition for promoting the production of the outer skin tissue according to the present invention promotes tyrosinase synthesis.
実施例８では、本発明に係る外皮系組織産生促進用組成物を含有する医薬品の一例として、外皮系組織産生促進用組成物含有軟膏を調製した。表５は、本実施例で調整した軟膏の配合組成を示す。 In Example 8, a composition-containing ointment for promoting skin tissue production was prepared as an example of a medicine containing the composition for promoting skin tissue production according to the present invention. Table 5 shows the formulation composition of the ointment prepared in this example.
実施例９では、本発明に係る外皮系組織産生促進用組成物を含有する飲食物の一例として、外皮系組織産生促進用組成物含有チューインガムを作製した。表６は、本実施例で作製したチューインガムの配合組成を示す。 In Example 9, a chewing gum containing a composition for promoting skin tissue production was prepared as an example of a food or drink containing the composition for promoting skin tissue production according to the present invention. Table 6 shows the composition of the chewing gum produced in this example.
実施例１０では、本発明に係る外皮系組織産生促進用組成物を含有する飲食物の一例として、外皮系組織産生促進用組成物含有クッキーを作製した。表７は、本実施例で作製したクッキーの配合組成を示す。 In Example 10, a composition-containing cookie for promoting skin tissue production was prepared as an example of a food or drink containing the composition for promoting skin tissue production according to the present invention. Table 7 shows the blended composition of the cookies produced in this example.
実施例１１では、本発明に係る外皮系組織産生促進用組成物を含有する飲食物の一例として、外皮系組織産生促進用組成物含有清涼飲料水を作製した。表８は、本実施例で作製した清涼飲料水の配合組成を示す。 In Example 11, as an example of a food or drink containing the composition for promoting the production of outer skin tissue according to the present invention, a soft drink containing the composition for promoting production of outer skin tissue was prepared. Table 8 shows the composition of the soft drink produced in this example.
実施例１２では、本発明に係る外皮系組織産生促進用組成物を含有する化粧品の一例として、外皮系組織産生促進用組成物含有化粧水を作製した。表９は、本実施例で作製した化粧水の配合組成を示す。 In Example 12, a composition-containing lotion for promoting skin tissue production was prepared as an example of a cosmetic containing the composition for promoting skin tissue production according to the present invention. Table 9 shows the composition of the lotion prepared in this example.
実施例１３では、本発明に係る外皮系組織産生促進用組成物を含有する化粧品の一例として、外皮系組織産生促進用組成物含有ヘアローションを作製した。表１０は、本実施例で作製したヘアローションの配合組成を示す。 In Example 13, a composition-containing hair lotion for promoting skin tissue production was prepared as an example of a cosmetic containing the composition for promoting skin tissue production according to the present invention. Table 10 shows the composition of the hair lotion produced in this example.
本発明に係る外皮系組織産生促進用組成物は、外皮系組織産生促進効果を有する。また、本発明に係る組成物は、天然由来成分であり、飲食物として摂取されたり、化粧品として利用されたりした実績があるため、長期に連続的に使用しても安全である可能性が高い。 The composition for promoting skin tissue production according to the present invention has an effect of promoting skin tissue production. In addition, the composition according to the present invention is a naturally-derived component and has a track record of being consumed as a food or drink or used as a cosmetic, so it is highly likely that it is safe to use continuously for a long period of time. .
本発明に係る外皮系組織産生促進用組成物を医薬品に含有させれば、種々の要因で引き起こされる脱毛症、白髪症、尋常性白斑症などの色素脱失症、先天性白皮症（アルビノ）等の予防及び治療に有効である。また、本発明に係る外皮系組織産生促進用組成物を飲食物（動物飼料含む）、又は化粧品に含有させれば、ヒト及び動物が容易に摂取することが可能となる。 If the composition for promoting the production of the integumental tissue according to the present invention is contained in a pharmaceutical product, depigmentation such as alopecia caused by various factors, baldness, vitiligo vulgaris, congenital albinism (albino ) And the like are effective for prevention and treatment. In addition, if the composition for promoting the production of the outer skin tissue according to the present invention is contained in foods and drinks (including animal feed) or cosmetics, humans and animals can easily take it.
１ 毛乳頭細胞
２ ヘアマトリックス（毛母基）
３ 外毛根鞘
４ 内毛根鞘
５ 立毛筋
６ 皮脂腺
７ 棍毛
８ 毛包バルジ領域
９ 新生（中）毛
1 Hair papillary cell 2 Hair matrix (hair matrix)
3 outer root sheath 4 inner root sheath 5 napped muscle 6 sebaceous gland 7 eyelash 8 hair follicle bulge region 9 new (middle) hair
Claims (4)
A cosmetic product for promoting melanin production comprising the melanin production promoter according to claim 1 or 2 as an active ingredient.
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| CN107243059A (en) * | 2017-06-16 | 2017-10-13 | 孙彦涛 | One kind treats albinic medicinal liquor prescription |
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| ES2420514B1 (en) * | 2012-02-22 | 2014-06-24 | Caroi'line Cosm�Tica S.L. | Erica erigena formulations and extracts to fight skin aging |
| JP6856223B2 (en) * | 2018-05-15 | 2021-04-07 | 株式会社ノムラ | Composition for human dermal papilla cell proliferation |
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| FR2669032B1 (en) * | 1990-11-14 | 1993-04-23 | Cecchi Georges | PROCESS FOR THE MANUFACTURE OF A PREPARATION OF D-ALPHA-TOCOPHEROL. |
| KR950007045B1 (en) * | 1992-04-14 | 1995-06-30 | 주식회사태평양 | Skin cosmetic material containing polyethoxylate vitamin e. |
| JPH0753334A (en) * | 1993-08-13 | 1995-02-28 | Monamuule:Kk | Hair restoring and growing cosmetic |
| JP3640727B2 (en) * | 1996-03-05 | 2005-04-20 | 呉羽化学工業株式会社 | Overcoat hair growth agent and hair growth method |
| FR2814950B1 (en) * | 2000-10-05 | 2003-08-08 | Oreal | USE OF AT LEAST ONE EXTRACT OF AT LEAST ONE PLANT FROM THE ERICACEAE FAMILY, IN COMPOSITIONS FOR TREATING SKIN SIGNS OF AGING |
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