KR102095031B1 - Composition for moisturing, anti-wrinkle or whitening with the extract of Zingiber mioga - Google Patents
Composition for moisturing, anti-wrinkle or whitening with the extract of Zingiber mioga Download PDFInfo
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- KR102095031B1 KR102095031B1 KR1020190114075A KR20190114075A KR102095031B1 KR 102095031 B1 KR102095031 B1 KR 102095031B1 KR 1020190114075 A KR1020190114075 A KR 1020190114075A KR 20190114075 A KR20190114075 A KR 20190114075A KR 102095031 B1 KR102095031 B1 KR 102095031B1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
Description
본 발명은 양하 추출물을 유효성분으로 하는 피부 보습, 피부 주름 개선 또는 피부 미백효과가 있는 식품 또는 화장료 조성물에 관한 것이다.The present invention relates to a food or cosmetic composition having moisturizing, skin wrinkle improvement or skin whitening effect using the extract of yangha as an active ingredient.
피부는 외부 환경으로부터 신체를 보호하는 역할을 하는 부위로 체온 조절, 감각 기능, 면역 기능 등을 수행한다. 이외에도 피부는 미용적인 측면을 담당하여 깨끗하고 건강한 피부는 미적 아름다움을 느끼게 하므로 피부 관련 시장이 활발하게 성장하고 있으며, 특히 나이가 들어감에 따라 나타나는 칙칙한 피부톤과 얼굴의 잔주름, 잃어가는 탄력 등의 현상으로 인해 소비자들은 피부 노화에 대한 관심이 높다.The skin is a part that protects the body from the external environment and performs body temperature control, sensory functions, and immune functions. In addition, the skin takes care of the cosmetic side, so the clean and healthy skin feels aesthetic beauty, so the skin-related market is actively growing, and in particular, it is caused by dull skin tone, fine wrinkles on the face, and loss of elasticity. Therefore, consumers are highly interested in skin aging.
탄력 있고, 잡티 없이 맑은 피부를 유지하기 위해서는 피부 보습 관리가 중요한데 피부 탄력에 관여하는 콜라겐 생성이 수분에 의해 좌우되기 때문이다. 피부의 수분이 부족하면, 세포의 재생능력이 저하되면서 탄력이 줄어들고 주름이 생기는 등의 노화 현상이 나타나게 된다. 이외에도 미백 관리, 영양 공급 등으로 건강한 피부를 유지할 수 있다.In order to maintain elastic and clear skin without blemishes, skin moisturizing is important because collagen production, which is involved in skin elasticity, is influenced by moisture. When the skin lacks moisture, aging occurs, such as reduced elasticity and wrinkles, as cells regenerate. In addition, healthy skin can be maintained by whitening management and nutrition.
한편, 피부 미백, 주름, 보습 등의 관리를 위해서 다양한 소재를 이용한 식품 또는 화장품이 연구 개발되고 있으며, 최근 화장품의 화학 성분으로 인해 부작용이 발생하는 사례가 빈번하면서, 천연물 유래 물질로 이루어진 제품에 대한 소비자의 관심이 갈수록 높아지고 있다.On the other hand, for the management of skin whitening, wrinkles, and moisturizing, foods or cosmetics using various materials are being researched and developed, and recently, there are frequent cases where side effects occur due to the chemical composition of cosmetics. Consumer interest is increasing.
종래에는 대한민국 등록특허 제10-1787074호(천연물 복합 추출물을 포함하는 피부 미백 및 피부 주름 개선용 화장료 조성물), 대한민국 공개특허 제10-2015-0059551호(인삼 새싹 추출물 또는 이의 분획물을 유효성분으로 함유하는 피부 상태 개선용 조성물 및 이를 이용한 피부 상태 개선 방법), 대한민국 공개특허 제10-2014-0058711호(백련근을 유효성분으로 함유하는 피부건강 개선용 식품 조성물 및 이를 포함한 한방건강식품) 등과 같이 천연물을 이용한 화장료 조성물 및 식품 조성물에 대한 기술을 제시한 바 있다.Conventionally, Korean Patent No. 10-1787074 (Cosmetic composition for skin whitening and skin wrinkle improvement including natural complex extract), Korean Patent No. 10-2015-0059551 (Ginseng sprout extract or a fraction thereof, as an active ingredient) Composition for improving skin condition and method for improving skin condition using the same), Korean Patent Publication No. 10-2014-0058711 (food composition for improving skin health containing baeknyeongeun as an active ingredient and herbal health food including the same) It has been proposed a technique for a cosmetic composition and a food composition using the.
본 발명은 피부 보습 및 피부 주름 개선 및 피부 미백효과가 뛰어난 천연물의 추출물을 발굴하고, 이를 유효성분으로 함유한 식품 또는 화장료 조성물을 개발하여 제공하는 것을 목적으로 한다.An object of the present invention is to discover an extract of a natural product excellent in skin moisturizing and skin wrinkle improvement and skin whitening effect, and to develop and provide a food or cosmetic composition containing it as an active ingredient.
본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 보습용 식품 조성물을 제공한다.The present invention provides a food composition for moisturizing the skin, characterized in that it contains a Yangha ( Zingiber mioga ) extract as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 주름 개선용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for improving skin wrinkles, characterized by containing Yangha ( Zingiber mioga ) extract as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 미백용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for skin whitening, characterized in that it contains an extract ( Zingiber mioga ) as an active ingredient.
한편, 본 발명에 있어서, 상기 양하 추출물은, 바람직하게 양하 꽃봉오리 추출물일 수 있다.On the other hand, in the present invention, the yangha extract, preferably may be yangha bud extract.
한편, 본 발명에 있어서, 상기 양하 추출물은, 바람직하게 양하 꽃봉오리 열수 추출물일 수 있다.On the other hand, in the present invention, the yangha extract, preferably may be yangha bud hot water extract.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 보습용 화장료 조성물을 제공한다.In addition, the present invention is Yang ( Zingiber mioga ) provides a cosmetic composition for moisturizing the skin, characterized in that it contains an extract as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 주름 개선용 화장료 조성물을 제공한다.In addition, the present invention is Yang ( Zingiber mioga ) It provides a cosmetic composition for improving skin wrinkles, characterized in that it contains an extract as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 미백용 화장료 조성물을 제공한다.In addition, the present invention is Yang ( Zingiber mioga ) It provides a cosmetic composition for skin whitening, characterized in that it contains an extract as an active ingredient.
한편, 본 발명에 있어서, 상기 양하 추출물은, 바람직하게 양하 꽃봉오리 추출물일 수 있다.On the other hand, in the present invention, the yangha extract, preferably may be yangha bud extract.
한편, 본 발명에 있어서, 상기 양하 추출물은, 바람직하게 양하 꽃봉오리 열수 추출물일 수 있다.On the other hand, in the present invention, the yangha extract, preferably may be yangha bud hot water extract.
본 발명은 양하 추출물을 유효성분으로 함유하는 식품 또는 화장료 조성물을 제공하여 뛰어난 피부 보습, 피부 주름 개선 및 피부 미백효과를 발휘할 수 있다.The present invention can provide excellent food moisturizing, skin wrinkle improvement and skin whitening effect by providing a food or cosmetic composition containing Yangha extract as an active ingredient.
도 1은 피부보습, 피부주름 및 피부미백에 관련된 각 세포의 생존능(viability)에 대한 양하 꽃봉오리 추출물의 영향을 나타내는 실험 결과 그래프이다.
도 2는 자외선(UVB)가 조사된 피부각질세포 HaCaT 세포 내 피부보습관련 유전자 HAS2와 Elastin의 mRNA 발현에 대한 양하 꽃봉오리 추출물의 영향을 나타내는 실험 결과 그래프이다. 실험에서 L-아르코르브산(L-ascorbic acid)을 양성 대조군으로 두었다.
도 3은 자외선(UVB)가 조사된 피부각질세포 HaCaT 세포 내 히알루론산 분비량에 대한 양하 꽃봉오리 추출물(A) 및 부위별 양하 추출물(꽃봉오리, 잎, 뿌리) (B)의 영향을 나타내는 실험 결과 그래프이다. 실험에서 L-아르코르브산(L-ascorbic acid)을 양성 대조군으로 두었다.
도 4의 A)는 자외선(UVB)가 조사된 섬유아세포 Hs27 세포 내 콜라겐 관련 유전자 프로콜라겐(Procollagen)의 mRNA 발현에 대한 양하 꽃봉오리 추출물의 영향을 나타낸 실험결과 그래프이다. 또한, 도 4의 B) 및 C)는 자외선(UVB)가 조사된 섬유아세포 Hs27 세포 내 콜라겐 관련 유전자 프로콜라겐(Procollagen)(B)과 Collagen type I (C)의 mRNA 발현에 대한 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)의 영향을 나타내는 실험 결과 그래프이다. 실험에서 L-아르코르브산(L-ascorbic acid)을 양성 대조군으로 두었다.
도 5는 자외선(UVB)가 조사된 섬유아세포 Hs27 세포 내 콜라겐 가수분해 관련 유전자 MMPs와 사이토카인 TNF-α의 mRNA 발현에 대한 양하 꽃봉오리 추출물의 영향을 나타내는 실험 결과 그래프이다. 실험에서 L-아르코르브산(L-ascorbic acid)을 양성 대조군으로 두었다.
도 6은 자외선(UVB)가 조사된 섬유아세포 Hs27 세포 내 염증성 사이토카인 IL-1β와 IL-6의 분비량에 대한 양하 꽃봉오리 추출물의 영향을 나타내는 실험 결과 그래프이다. 실험에서 L-아르코르브산(L-ascorbic acid)을 양성 대조군으로 두었다.
도 7은 멜라닌 생성세포 B16F10의 세포의 티로시나제(Tyrosinase) 활성에 대한 양하 꽃봉오리 추출물 (A) 및 부위별 양하 추출물(꽃봉오리, 잎, 뿌리) (B)의 영향을 나타내는 실험 결과 그래프이다. 실험에서 알부틴(Arbutin)을 양성 대조군으로 두었다.
도 8은 cAMP를 자극하는 IBMX-유도된 멜라닌 생성세포 B16F10 내 멜라닌 생성에 대한 양하 꽃봉오리 추출물의 영향을 나타내는 실험 결과 그래프이다. 실험에서 알부틴(Arbutin)을 양성 대조군으로 두었다.1 is a graph showing the results of experiments showing the effect of the extract of yangha bud on the viability of each cell related to skin moisturizing, skin wrinkles and skin whitening.
Figure 2 is a graph showing the results of experiments showing the effect of the extract of yangha bud on the mRNA expression of the skin moisturizing genes HAS2 and Elastin in the skin keratinocytes HaCaT cells irradiated with ultraviolet light (UVB). In the experiment, L-ascorbic acid was set as a positive control.
3 is an experimental result showing the effect of yangha bud extract (A) and yangha extract (buds, leaves, roots) (B) on the secretion of hyaluronic acid in the skin keratinocytes HaCaT cells irradiated with ultraviolet (UVB) It is a graph. In the experiment, L-ascorbic acid was set as a positive control.
A of FIG. 4 is a graph showing the results of experiments showing the effect of the extract of yangha bud on the mRNA expression of collagen-related gene procollagen in Hs27 cells with UV (UVB) irradiation. In addition, B) and C) of FIG. 4 are extracts by region for mRNA expression of collagen-related genes Procollagen (B) and Collagen type I (C) in fibroblast Hs27 cells irradiated with ultraviolet light (UVB). It is a graph of the experimental results showing the effect of (bud, leaf, root). In the experiment, L-ascorbic acid was set as a positive control.
5 is a graph showing the results of experiments showing the effect of the extract of yangha bud on the mRNA expression of collagen hydrolysis-related gene MMPs and cytokine TNF-α in ultraviolet (UVB) irradiated fibroblast Hs27 cells. In the experiment, L-ascorbic acid was set as a positive control.
FIG. 6 is a graph showing the results of experiments showing the effect of extracts of yangha buds on the secretion of inflammatory cytokines IL-1β and IL-6 in UVB-irradiated fibroblast Hs27 cells. In the experiment, L-ascorbic acid was set as a positive control.
FIG. 7 is a graph showing the results of experiments showing the effect of a yangha bud extract (A) and a yangha extract (bud, leaf, root) (B) on the tyrosinase activity of melanin producing cells B16F10 cells. Arbutin was used as a positive control in the experiment.
8 is a graph showing the results of experiments showing the effect of the extract of yangha bud on the production of melanin in IBMX-induced melanin producing cells B16F10 that stimulate cAMP. Arbutin was used as a positive control in the experiment.
양하(Zingiber mioga)는 아시아 열대 지방을 원산지로 둔 생강과의 여러해살이풀로, 독특한 향이 있어 향미채소로 활용되거나 생으로 섭취하기도 한다. 국내에서는 전국 산지나 들에서 접할 수 있으며 특히 남부지방과 제주도에서 즐겨먹는 채소이다. 양하는 제주도에서 '양왜', '양애'로 부르며, 전남지방에서는 '양해'로 부르며 일본에서는 '묘가'라고 부르고, 일본의 대중적인 식재료로 애용된다.Yang Ha ( Zingiber mioga ) is a perennial plant with ginger originating from tropical Asia, and has a unique scent that is used as a flavor vegetable or consumed raw. In Korea, it can be found in the mountains and fields of the country. It is a vegetable especially enjoyed in southern regions and Jeju Island. Yangha is called 'Yang Wa' and 'Yang Ae' in Jeju Island, 'Yanghae' in Jeonnam Province, and 'Myoga' in Japan, and is used as a popular food ingredient in Japan.
양하는 칼륨, 칼슘, 마그네슘, 철분 등의 무기질이 다양하게 함유되어 있으며, 한방에서는 뿌리줄기와 종자를 약재로 사용하여 여성의 생리불순과 백대화 치료 및 진해, 거담 등을 해결하는데 효과를 가지는 것으로 알려져 있다. 또한, 줄기로 국을 끓여 먹거나 연한 잎사귀로 쌈을 싸 먹기도 하고, 어린순과 뿌리는 향신료로 이용되기도 한다. Yangha contains various minerals such as potassium, calcium, magnesium, and iron, and in oriental medicine, it has the effect of treating rhizome and white talk, and treating coughing and expectoration of women by using rhizomes and seeds as medicines. Is known. In addition, boiled soup with stalks or wrapped in ssam with light leaves, and sprouts and roots are also used as spices.
본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 보습용 식품 또는 화장료 조성물을 제공한다.The present invention provides a food or cosmetic composition for skin moisturizing, characterized in that it contains an extract of Yangha ( Zingiber mioga ) as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 주름 개선용 식품 또는 화장료 조성물을 제공한다.In addition, the present invention is Yang ( Zingiber mioga ) It provides a food or cosmetic composition for improving skin wrinkles, characterized in that it contains an extract as an active ingredient.
또한, 본 발명은 양하(Zingiber mioga) 추출물을 유효성분으로 함유하는 것을 특징으로 하는 피부 미백용 식품 또는 화장료 조성물을 제공한다.In addition, the present invention provides a food or cosmetic composition for skin whitening, characterized by containing Yangha ( Zingiber mioga ) extract as an active ingredient.
본 발명의 조성물에 있어서, 상기 양하(Zingiber mioga) 추출물은 양하의 꽃봉오리 추출물인 것이 바람직하다. 여기서, 꽃봉오리는 망울만 맺혀 아직 피지 않은 꽃을 의미하는 것으로, 꽃눈이 상당히 발당하여 개화가 가까워진 상태를 의미한다. 양하의 꽃봉오리는 양하의 다른 부위(예를 들면, 줄기, 잎, 뿌리 등)나 활짝 핀 꽃에 비하여 증진된 피부 보습 효과, 피부 주름 개선 효과와 피부 미백 효과를 발휘할 수 있다. 이에 본원발명은 양하 꽃봉오리 추출물을 이용하여 피부 보습 효과, 피부 주름 개선 효과, 피부 미백 효과를 발휘하는 식품 조성물이나 화장료 조성물을 제공할 수 있으며, 피부 보습 효과, 피부 주름 개선 효과와 피부 미백 효과를 복합적으로 발휘할 수 있는 복합 기능성 식품 조성물이나 화장료 조성물을 제공할 수 있다. In the composition of the present invention, the yangha ( Zingiber mioga ) extract is preferably a bud extract of Yangha. Here, the flower bud means a flower that has not yet bloomed because only the buds are formed. Yangha's flower buds can exert enhanced skin moisturizing effects, skin wrinkle improvement and skin whitening effects compared to other parts of Yangha (eg, stems, leaves, roots, etc.) or blooming flowers. Accordingly, the present invention can provide a food composition or a cosmetic composition that exerts a skin moisturizing effect, a skin wrinkle improving effect, and a skin whitening effect using a Yangha bud extract, and has a skin moisturizing effect, a skin wrinkle improving effect and a skin whitening effect. It is possible to provide a composite functional food composition or cosmetic composition capable of being exerted in a complex manner.
구체적으로, 본 발명의 양하 꽃봉오리 추출물은 피부각질세포인 HaCaT 세포에서 피부 보습 관련 유전자 HAS2와 Elastin의 발현을 증가시키고, 또한 히알루론산의 분비량을 증가시켜 피부 보습 효과를 발휘할 수 있다.Specifically, the yangha bud extract of the present invention can increase the expression of the skin moisturizing related genes HAS2 and Elastin in HaCaT cells, which are skin keratinocytes, and also increase the secretion of hyaluronic acid to exert a skin moisturizing effect.
또한, 본 발명의 양하 꽃봉오리 추출물은 콜라겐 관련 유전자 Procollagen과 Collagen type I의 발현량을 증가시키고, 콜라겐 가수분해 관련 유전자 MMPs와 염증성 사이토카인 TNF-α의 발현량을 감소시키며, 염증성 사이토카인인 IL-1β 및 IL-6의 분비량을 감소시켜, 피부 주름 개선 효과를 발휘할 수 있다.In addition, the yangha bud extract of the present invention increases the expression level of the collagen-related genes Procollagen and Collagen type I, decreases the expression level of the collagen hydrolysis-related gene MMPs and the inflammatory cytokine TNF-α, and the inflammatory cytokine IL By reducing the secretion of -1β and IL-6, it can exert the effect of improving skin wrinkles.
또한, 본 발명의 양하 꽃봉오리 추출물은 멜라닌 생성세포인 B16F10 세포에서 멜라닌 생성량을 감소시키고, 멜라닌 생성에 관여하는 효소 티로시나제 활성을 억제시켜, 피부 미백 효과를 발휘할 수 있다.In addition, the yangha bud extract of the present invention may reduce melanin production in B16F10 cells, which are melanin-producing cells, and inhibit the enzyme tyrosinase activity involved in melanin production, thereby exerting skin whitening effects.
본 발명의 조성물에 있어서, 상기 양하 꽃봉오리 추출물은 본 발명이 속하는 기술 분야에서 통상적으로 사용되는 다양한 추출 방법을 이용할 수 있다. 예를 들면, 정제수, 탄소수 1-4의 무수 또는 함수 저급 알코올, 프로필렌글리콜, 부틸렌글리콜, 글리세린, 아세톤, 에틸아세테이트, 클로로포름, 부틸아세테이트, 디에틸에테르, 디클로로메탄, 헥산 및 이들의 혼합물로 구성된 군으로부터 선택되는 추출용매를 이용한 용매 추출법, 초임계 추출법 및 초음파 추출법 중 선택되는 어느 하나의 추출법을 통해 수득될 수 있다. In the composition of the present invention, the yangha bud extract can use various extraction methods commonly used in the art. For example, consisting of purified water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, dichloromethane, hexane, and mixtures thereof. It can be obtained through any one extraction method selected from a solvent extraction method, a supercritical extraction method and an ultrasonic extraction method using an extraction solvent selected from the group.
본 발명에서 양하 꽃봉오리 추출물은 바람직하게, 양하 꽃봉오리에 정제수를 가하여 추출한 후, 감압농축하고 동결건조하여 수득된 것을 사용하는 것이 좋다. 예를 들면, 양하 꽃봉오리 100 내지 400중량부에 대하여 정제수 1200 내지 1800 중량부를 첨가한 후, 80 내지 90℃에서 6시간 내지 10시간 추출하여 양하 추출액을 수득하고, 그 후, 여과된 추출액을 50 내지 60℃에서 감압 농축한 다음 동결건조하여 얻은 양하 꽃봉오리 열수 추출물을 사용하는 것이 좋다.Yangha bud extract in the present invention, preferably, after adding purified water to the Yangha bud, extract, it is preferable to use the one obtained by concentration under reduced pressure and freeze-drying. For example, after adding 1200 to 1800 parts by weight of purified water with respect to 100 to 400 parts by weight of Yangha buds, extraction was performed at 80 to 90 ° C for 6 to 10 hours to obtain a sake extract, and thereafter, the filtered extract was 50. It is preferable to use a hot water extract of yangha bud obtained by concentration under reduced pressure at 60 ° C. and then freeze-dried.
한편, 본 발명의 식품 조성물은 일 예로, 육류, 곡류, 카페인 음료, 일반음료, 초콜렛, 빵류, 스넥류, 과자류, 피자, 젤리, 면류, 껌류, 아이스크림류, 알코올성 음료, 술, 비타민 복합제 및 그 밖의 건강보조식품류 중 선택되는 어느 하나일 수 있으며, 반드시 이에 한정되는 것은 아니다.On the other hand, the food composition of the present invention, for example, meat, cereals, caffeine drinks, general drinks, chocolate, bread, snacks, confectionery, pizza, jelly, noodles, gums, ice cream, alcoholic beverages, alcohol, vitamin complexes and other It may be any one selected from health supplements, and is not necessarily limited thereto.
한편, 본 발명의 화장료 조성물은, 일 예로, 용액, 현탁액, 유탁액, 페이스트, 화장수, 젤, 수용성 리퀴드, 크림, 에센스, 계면활성제-함유 클렌징, 오일, 수중유(O/W)형 및 유중수(W/O)형 중 선택되는 어느 하나의 기초화장료 제형; 스킨; 로션; 아이크림; 수딩젤; 연고; 마스크팩용 제형; 바디워시용 제형; 필링젤; 수중유형 및 유중수형 메이크업베이스; 파운데이션; 스킨커버; 립스틱, 립그로스, 페이스파우더, 투웨이케익, 아이새도, 치크칼라 및 아이브로우 펜슬류 중 선택되는 어느 하나의 색조화장료 제형; 두피용 제형; 중에서 선택되는 어느 하나인 것일 수 있다.On the other hand, the cosmetic composition of the present invention, for example, solution, suspension, emulsion, paste, lotion, gel, water-soluble liquid, cream, essence, surfactant-containing cleansing, oil, oil-in-water (O / W) type and oil Any one of the basic cosmetic formulations selected from heavy water (W / O) type; skin; Lotion; Eye cream; Soothing gel; Ointment; Formulation for mask packs; Body wash formulations; Peeling gel; Water-in-oil and water-in-oil makeup bases; foundation; Skin cover; Lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color, and eyebrow pencils; Scalp formulations; It may be any one selected from.
또한, 본 발명의 화장료 조성물은 본 발명 이외의 다른 화장료 조성물과 중복하여 사용할 수 있다. 또한 본 발명에 따른 화장료 조성물은 통상적인 사용방법에 따라 사용될 수 있으며, 사용자의 피부 상태 또는 취향에 따라 그 사용횟수를 달리할 수 있다.In addition, the cosmetic composition of the present invention can be used in combination with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention may be used according to a conventional method of use, and the number of times of use may be varied according to a user's skin condition or taste.
이하, 본 발명에 대해 하기 실시예 및 실험예에서 더욱 상세히 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 이와 등가의 기술적 사상의 변형까지를 모두 포함한다.Hereinafter, the present invention will be described in more detail in the following Examples and Experimental Examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes all modifications of equivalent technical ideas.
[[ 실시예Example : 본 발명의 양하 추출물의 제조] : Preparation of Yangha extract of the present invention]
본 실시예에서는 하기와 같이 양하 부위에 따라 양하 추출물을 제조하였다.In this example, extracts of yangha were prepared according to yangha as shown below.
1) 양하 꽃봉오리 추출물의 제조1) Preparation of Yangha flower bud extract
양하 꽃봉오리 200kg에 정제수 1600L를 넣고 85℃에서 8시간 추출하여 양하 추출액을 수득하였다. 그 후, 여과된 추출액을 60℃ 이하에서 감압농축한 다음 동결건조하여 양하 꽃봉오리 추출물을 제조하였다. 하기 실험에서는 이를 FSH-ZM으로 표기하였다. 1600 L of purified water was added to 200 kg of Yangha buds, and extracted at 85 ° C. for 8 hours to obtain a Yangha extract. Subsequently, the filtered extract was concentrated under reduced pressure at 60 ° C. or less, and then freeze-dried to prepare a yangha bud extract. In the following experiment, this was designated as FSH-ZM.
2) 양하 잎 추출물의 제조2) Preparation of Yangha leaf extract
양하 잎 50kg에 정제수 8배수를 넣고 85℃에서 8시간 추출하여 양하 추출액을 수득하였다. 그 후, 여과된 추출액을 60℃ 이하에서 감압농축한 다음 동결건조하여 양하 잎 추출물을 제조하였다. 하기 실험에서는 이를 FSH-ZM(L)으로 표기하였다. 8 kg of purified water was added to 50 kg of the leaves of yangha and extracted at 85 ° C. for 8 hours to obtain a yangha extract. Subsequently, the filtered extract was concentrated under reduced pressure at 60 ° C. or lower, and then freeze-dried to prepare a leaf extract. In the following experiment, this was designated as FSH-ZM (L).
3) 양하 뿌리 추출물의 제조3) Preparation of Yangha Root Extract
양하 뿌리 50kg에 정제수 8배수를 넣고 85℃에서 8시간 추출하여 양하 추출액을 수득하였다. 그 후, 여과된 추출액을 60℃ 이하에서 감압농축한 다음 동결건조하여 양하 뿌리 추출물을 제조하였다. 하기 실험에서는 이를 FSH-ZM(R)으로 표기하였다. 8 kg of purified water was added to 50 kg of the roots of yangha and extracted at 85 ° C for 8 hours to obtain a yangha extract. Subsequently, the filtered extract was concentrated under reduced pressure at 60 ° C. or less, and then freeze-dried to prepare a yangha root extract. In the following experiment, this was designated as FSH-ZM (R).
하기에서는 상기 실시예의 양하 추출물을 이용하여 피부 보습, 피부 주름, 피부 미백에 대한 효과를 확인하기 위한 실험을 진행하였으며, 이때, 양성대조군으로는 L-아스코르브산(L-ascorbic acid)과 알부틴(Arbutin)을 사용하였으며 Sigma-Aldrich Co.(St. Louis, MO, USA)에서 구입하여 실험에 사용하였다. 하기 실험은 SPSS(Statis tical Package for the Social Science) version 22 프로그램을 이용하여 분석하였고, 모든 측정 결과는 평균(mean)±표준편차(Standard deviation, SD)로 표시하였다. 또한, 그룹간의 통계적 유의성을 Duncan's multiple range test를 실시하였으며 p<0.05 수준에서 유의성의 여부를 검증하였다.In the following, experiments were carried out to confirm the effects on skin moisturizing, skin wrinkles, and skin whitening using the extract of the extract of the above example, and at this time, L-ascorbic acid and arbutin were positive controls. ) And was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and used in the experiment. The following experiment was analyzed using the SPSS (Statis tical Package for the Social Science) version 22 program, and all measurement results were expressed as the mean ± standard deviation (SD). In addition, Duncan's multiple range test was performed for statistical significance between groups, and the significance was verified at p <0.05.
[실험예 1 : 양하 추출물의 피부 보습 효과 확인][Experimental Example 1: Confirming the skin moisturizing effect of Yangha extract]
본 실험예에서는 상기 실시예에서 제조한 양하 추출물의 피부 보습에 대한 효과를 확인하고자 하였다.In this experimental example, the effect of the extract of yangha prepared in the above example on skin moisturization was intended.
1) 세포 배양 1) Cell culture
실험에 사용된 피부각질세포인 HaCaT는 American Type Cultured Collection (ATCC; Rockville, MD, USA)에서 분주 받아 실험하였다. 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (100 units/mL)이 함유된 High-glucose Dulbeco’s modified Eagle’s medium 배지를 사용하였고, 37℃, 5% CO₂, 95% humid air로 조절된 배양기(Thermo Fisher Scientific Inc., Pittsburgh, PA, USA)에서 배양하였다.The skin keratinocytes used in the experiment, HaCaT, was tested by dispensing from the American Type Cultured Collection (ATCC; Rockville, MD, USA). High-glucose Dulbeco's modified Eagle's medium medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (100 units / mL) was used, and incubator adjusted to 37 ° C, 5% CO2, and 95% humid air (Thermo Fisher Scientific Inc., Pittsburgh, PA, USA).
2) 세포 독성 시험2) Cytotoxicity test
HaCaT 세포를 96-well plate에 1×104 cells/well로 분주하여 24시간 동안 배양한 후, 각 well에 양하 꽃봉오리 추출물을 농도별로 처리하였다. 24시간 후, 5 mg/ml 농도의 MTT solution을 20 μL 첨가하고 4시간 동안 배양한 후, 배지를 제거하고 DMSO 100 μL/well 씩 첨가하여 560 nm에서 흡광도를 측정하였다.HaCaT cells were dispensed into a 96-well plate at 1 × 10 4 cells / well and cultured for 24 hours, and then, each well was treated with extracts of yangha buds by concentration. After 24 hours, after adding 20 μL of 5 mg / ml MTT solution and incubating for 4 hours, the medium was removed and DMSO was added at 100 μL / well to measure absorbance at 560 nm.
그 결과, 도 1의 (A)와 같이 양하 꽃봉오리 추출물 200 μg/mL 이하의 농도에서 세포독성이 나타나지 않는 것을 확인하였으며, 이에 본 실험에서는 양하 꽃봉오리 추출물 200μg/mL 농도까지 처리하였다.As a result, as shown in Figure 1 (A), it was confirmed that the cytotoxicity does not appear at a concentration of 200 μg / mL or less of the yangha bud extract, and in this experiment, the yangha bud extract was treated to a concentration of 200 μg / mL.
3) 양하 꽃봉오리 추출물의 피부 보습 관련 유전자 발현량 측정 실험3) Experiment of measuring gene expression level related to skin moisturization of Yangha bud extract
상기 실시예의 양하 꽃봉오리 추출물이 HaCaT 세포에서 자외선 조사에 의한 보습 관련 유전자들의 발현량에 미치는 영향을 확인하기 위해 HaCaT 세포의 RNA 추출 후, 실시간 정량 PCR(Real-time PCR)을 진행하였다.After confirming the effect of the extract of the buds of the above Example on the expression level of moisturizing-related genes by UV irradiation in HaCaT cells, RNA extraction of HaCaT cells was performed, followed by real-time quantitative PCR (Real-time PCR).
먼저 HaCaT 세포를 배양하여 6 well plate에 2×105 cells/well 분주한 후 24시간 배양하였다. 배지를 제거한 후 DPBS로 세척하고 UVB lamp(5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan)를 이용하여 50 mJ/㎠ 조사한 후 양성대조군인 L-ascorbic acid 및 양하 꽃봉오리 추출물을 50, 100, 200 μg/mL 농도로 처리하여 24시간 배양하였다. 배양한 HaCaT 세포의 RNA 추출은 RNeasy Mini kit (Qiagen, Valencia, CA, USA)를 이용하였고, iScript cDNA Synthesis kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA)로 cDNA를 합성하였다. First, HaCaT cells were cultured, 2 × 10 5 cells / well were dispensed into 6 well plates, and then cultured for 24 hours. After removing the medium, washed with DPBS, irradiated with UVB lamp (5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan) and irradiated with 50 mJ / ㎠, the positive control L-ascorbic acid and
피부 보습 관련 유전자들의 발현을 측정하기 위하여 SYBR Green (iQ SYBR Green Supermix, Bio-Rad Laboratories Inc.)을 이용한 Real-Time PCR (Bio-Rad Laboratories, Inc., Hercules, CA, USA)을 이용하였다. 각각의 유전자에 대한 PCR primer의 염기서열은 하기 표 1에 나타내었다. Real-time PCR 반응은 총 20 μL 내에 10 μL의 2X SYBR mix, 2 μL의 cDNA, primer는 각각 100 pmol/μL를 1 μL씩 첨가하였고 나머지는 nuclease free water로 채워주었다. 모든 유전자에 대하여 PCR 증폭 단계는 다음과 같고 증폭 cycle은 40 cycle을 실시하였다. Hot start를 위해 95℃에서 10분, Denaturation을 95℃에서 15초, annealing을 58℃에서 30초, extension을 72℃에서 30초간 반복하며 각 cycle의 extension 후에 값이 기록되었다. 모든 cycle이 완료된 후 primer의 특이성을 확인하기 위해 melting curve 분석을 실시하였다. mRNA의 상대적인 함량은 자외선 처리를 하지 않은 군에 대한 상대적인 양으로 나타내었다.Real-Time PCR (Bio-Rad Laboratories, Inc., Hercules, CA, USA) using SYBR Green (iQ SYBR Green Supermix, Bio-Rad Laboratories Inc.) was used to measure the expression of skin moisturizing related genes. The base sequence of PCR primers for each gene is shown in Table 1 below. In the real-time PCR reaction, 1 μL of 100 pmol / μL was added to each of 10 μL of 2X SYBR mix, 2 μL of cDNA, and primer in 20 μL, and the rest was filled with nuclease free water. PCR amplification steps for all genes were as follows, and the amplification cycle was 40 cycles. For the hot start, the value was recorded after the extension of each cycle by repeating for 10 minutes at 95 ° C, Denaturation at 95 ° C for 15 seconds, annealing at 58 ° C for 30 seconds, and extension at 72 ° C for 30 seconds. After all cycles were completed, melting curve analysis was performed to confirm the specificity of the primer. The relative content of mRNA was expressed as a relative amount to the group not treated with ultraviolet rays.
진피에는 섬유아세포가 존재하는데, 섬유아세포는 결합조직으로 탄력을 책임지는 성분인 콜라겐(Collagen)과 엘라스틴(Elastin)을 생산하며, 두 성분 사이에는 진피 내 보습을 책임지는 성분인 히알루론산(Hyaluronic Acid, HA)이 채워져 있다.따라서 콜라겐, 엘라스틴 및 히알루론산이 부족하면 비어 있는 공간이 많아 탄력 및 보습력이 떨어지게 된다. Fibroblasts are present in the dermis. Fibroblasts produce collagen and elastin, the components responsible for elasticity in connective tissue, and hyaluronic acid, a component responsible for moisturizing in the dermis, between the two components. , HA). Therefore, when collagen, elastin and hyaluronic acid are insufficient, there are many empty spaces, resulting in poor elasticity and moisturizing ability.
이와 관련한 유전자들의 발현량을 확인한 결과, 도 2의 (A)와 같이 히알루론산 합성효소인 HAS2의 유전자 발현량은 Normal control 군에 비해 UVB를 조사한 control군에서의 발현량이 유의적으로 감소한 반면, 양하 꽃봉오리 추출물 처리군의 경우 control 군에 비해 농도 의존적으로 증가하여 유의성이 나타났다. 특히, 양하 꽃봉오리 추출물 처리군의 100과 200 μg/mL 농도에서 control 군 대비 각각 80%, 152.7% 유의적 증가를 확인하였다.As a result of confirming the expression level of the genes related to this, as shown in FIG. 2 (A), the expression level of the hyaluronic acid synthetase HAS2 was significantly decreased in the control group irradiated with UVB compared to the normal control group, while lowering In the case of the bud extract treatment group, the concentration was significantly increased compared to the control group, indicating a significance. In particular, it was confirmed that the concentration of 100% and 200 μg / mL of the Yangha bud extract treatment group was significantly increased by 80% and 152.7%, respectively, compared to the control group.
또한, 도 2의 (B)와 같이 Elastin의 유전자 발현량은 Normal control군에 비해 control 군에서의 발현량이 유의적으로 감소한 반면, 양하 꽃봉오리 추출물 처리군의 경우 농도 의존적으로 증가하는 경향을 보이며, control군에 비해 200 μg/mL 농도에서 67.8% 유의적 증가를 나타냈다. In addition, as shown in FIG. 2 (B), the expression level of Elastin was significantly decreased in the control group compared to the normal control group, whereas in the case of the yangha bud extract treatment group, it showed a tendency to increase in a concentration-dependent manner, It showed a 67.8% significant increase in the concentration of 200 μg / mL compared to the control group.
4) 양하 추출물이 히알루론산 분비량에 미치는 영향 확인 실험4) Experiment to confirm the effect of Yangha extract on hyaluronic acid secretion
상기 실시예의 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)이 HaCaT 세포에서 자외선 조사에 의한 히알루론산 분비량에 미치는 영향을 확인하기 위해 하기와 같은 실험을 진행하였다. The following experiments were performed to confirm the effect of the extracts of each part by region (buds, leaves, and roots) on the amount of hyaluronic acid secretion by UV irradiation in HaCaT cells.
HaCaT 세포를 배양하여 96 well plate에 1×104 cells/well 분주한 후 24시간 배양하였다. 배지를 제거한 후 DPBS로 세척하고 UVB lamp(5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan)를 이용하여 50 mJ/㎠조사한 후 양성대조군인 L-ascorbic acid 및 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)을 50, 100, 200 μg/mL 농도로 처리하여 24시간 배양하였다. 상층액을 분리하여 Hyaluronan 분비량 측정에 있어서 시료로 사용하였다. 새로운 96 well plate에 50 μL의 assay diluent를 분주하고 standard와 시료를 50 μL씩 첨가하여 cover를 씌워 2시간 동안 실온에서 shaking 하였다. 96 well plate를 washing 한 후 well마다 100 μL의 hyaluronan conjugate를 분주한 후 빛을 차단하여 반응시킨 후 ELISA reader (Bio-Rad Laboratories Headquarters, Hercules, CA, USA)로 655 nm에서 흡광도를 측정하였다.HaCaT cells were cultured, 1 × 10 4 cells / well were dispensed into a 96 well plate, and cultured for 24 hours. After removing the medium, washed with DPBS, irradiated with 50 mJ / ㎠ using a UVB lamp (5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan), and positive control L-ascorbic acid and extracts by region (buds) , Leaves, roots) were cultured at 50, 100, 200 μg / mL concentration for 24 hours. The supernatant was separated and used as a sample in the measurement of Hyaluronan secretion. 50 μL of assay diluent was dispensed into a new 96 well plate, and 50 μL of standard and samples were added, and the cover was covered and shaken at room temperature for 2 hours. After washing the 96-well plate, 100 μL of hyaluronan conjugate was dispensed for each well, and then the light was blocked to react and the absorbance was measured at 655 nm with an ELISA reader (Bio-Rad Laboratories Headquarters, Hercules, CA, USA).
히알루론산은 주로 각질형성세포에서 형성되며 다당류가 많이 들어있어 수분을 상당량 함유할 수 있는 특징이 있는 인자로 알려져 있다. 자외선으로 손상된 HaCaT cell에 양하 꽃봉오리 추출물을 처리하여 히알루론산의 분비량을 측정한 결과, 도 3)의 A)와 같이 Normal control군에 비해 UVB를 조사한 군들의 히알루론산의 분비량이 상당히 감소하였음을 확인하였고, 유의성이 나타났다. 양하 꽃봉오리 추출물 처리군의 경우 UVB조사로 감소된 히알루론산의 분비량을 control 군에 비해 50, 100, 200 μg/mL농도에서 각각 2.6%, 6.5%, 10.6%로 농도 의존적으로 증가하는 것을 확인하였다.Hyaluronic acid is mainly formed in keratinocytes and contains many polysaccharides, so it is known as a factor that can contain a significant amount of moisture. As a result of measuring the secretion amount of hyaluronic acid by treating the HaCaT cell damaged by ultraviolet rays, the secretion amount of hyaluronic acid was significantly reduced in the groups irradiated with UVB compared to the normal control group as shown in Fig. 3) A). And significance. In the case of the Yangha bud extract treatment group, it was confirmed that the secretion amount of hyaluronic acid decreased by UVB irradiation increased concentration-dependently at concentrations of 50, 100, and 200 μg / mL to 2.6%, 6.5%, and 10.6%, respectively, compared to the control group. .
또한, 자외선으로 손상된 HaCaT cell에 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)을 처리하여 히알루론산의 분비량을 측정한 결과, 도 3의 B)와 같이 Normal control군에 비해 UVB를 조사한 군들의 히알루론산의 분비량이 상당히 감소하였음을 확인하였고, 유의성이 나타났다. 부위별 양하 추출물 처리군의 경우 control 군에 비해 UVB조사로 감소된 히알루론산의 분비량이 증가하였고(각 26%, 10%, 4%), 특히 양하 꽃봉오리 추출물(FSH-ZM)에서 유의적으로 가장 높은 증가를 보이는 것을 확인하였다.In addition, as a result of measuring the secretion amount of hyaluronic acid by treating the HaCaT cell damaged by ultraviolet rays by extracts (buds, leaves, and roots) for each region, as shown in FIG. 3B), hyalurism of the groups irradiated with UVB compared to the normal control group It was confirmed that the amount of lactic acid secretion was significantly reduced, and significance was shown. Compared with the control group, the amount of hyaluronic acid secretion decreased by the UVB irradiation in the case of the extract treatment group by region (26%, 10%, 4%, respectively), especially in the Yangha bud extract (FSH-ZM). It was confirmed that it showed the highest increase.
이와 같이 피부각질세포인 HaCaT 세포에서 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)은 히알루론산의 분비량을 control군에 비해 증가시켰고, 특히 양하 꽃봉오리 추출물은 피부 보습 관련 유전자 HAS2와 Elastin의 발현을 증가시키며, control군에 비해 히알루론산의 분비량을 증가시키면서 다른 부위별 양하 추출물(잎, 뿌리)에 비해서도 그 증가량이 높은 것을 확인하였다. 이를 통해 양하 꽃봉오리를 피부 보습에 효능이 있는 물질로 이용할 수 있음을 알 수 있었다.As described above, the extracts of yangha (buds, leaves, and roots) by region in the skin keratinocytes, HaCaT cells, increased the secretion of hyaluronic acid compared to the control group. It increased, and increased the amount of hyaluronic acid secretion compared to the control group. Through this, it was found that the Yangha flower bud can be used as a substance effective for moisturizing the skin.
[실험예 2 : 양하 추출물의 피부 주름 개선 효과 확인][Experimental Example 2: Confirming the effect of Yangha extract on skin wrinkle improvement]
본 실험예에서는 상기 실시예에서 제조한 양하 추출물의 피부 주름 개선 효과를 확인하고자 하였다.In this experimental example, it was intended to confirm the effect of improving the skin wrinkles of the extract of yangha prepared in the above example.
1) 세포 배양 1) Cell culture
실험에 사용된 인간섬유아세포인 Hs27은 American Type Cultured Collection (ATCC; Rockville, MD, USA)에서 분주 받아 실험하였다. 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (100 units/mL)이 함유된 High-glucose Dulbeco’s modified Eagle’s medium배지를 사용하였고, 37℃, 5% CO₂, 95% humid air로 조절된 배양기(Thermo Fisher Scientific Inc., Pittsburgh, PA, USA)에서 배양하였다.The human fibroblast used in the experiment, Hs27, was tested by dispensing from the American Type Cultured Collection (ATCC; Rockville, MD, USA). High-glucose Dulbeco's modified Eagle's medium medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (100 units / mL) was used, and incubator adjusted to 37 ℃, 5% CO₂, 95% humid air (Thermo Fisher Scientific Inc., Pittsburgh, PA, USA).
2) 세포 독성 시험2) Cytotoxicity test
Hs27 세포를 96-well plate에 1×104 cells/well로 분주하여 24시간 동안 배양한 후, 각 well에 양하 꽃봉오리 추출물을 농도별로 처리하였다. 24시간 후, 5 mg/ml 농도의 MTT solution을 20 μL 첨가하고 4시간 동안 배양한 후, 배지를 제거하고 DMSO 100 μL/well 씩 첨가하여 560 nm에서 흡광도를 측정하였다.After dispensing Hs27 cells in a 96-well plate at 1 × 10 4 cells / well and incubating for 24 hours, each well was treated with extracts of yangha buds by concentration. After 24 hours, after adding 20 μL of 5 mg / ml MTT solution and incubating for 4 hours, the medium was removed and DMSO was added at 100 μL / well to measure absorbance at 560 nm.
그 결과, 도 1의 (B)와 같이 양하 꽃봉오리 추출물 200 μg/mL이하의 농도에서 세포독성이 나타나지 않는 것을 확인하여 본 실험에서는 양하 꽃봉오리 추출물 200 μg/mL농도까지 처리하였다.As a result, as shown in (B) of FIG. 1, it was confirmed that cytotoxicity was not observed at a concentration of 200 μg / mL or less of the extract of the buds of the yangha, and to the concentration of 200 μg / mL of the extracts of the yangha of the buds.
3) 양하 추출물의 피부 주름 관련 유전자 발현량 측정 실험3) Measurement of gene expression level related to skin wrinkles of Yangha extract
상기 실시예의 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)이 Hs27 세포에서 자외선 조사에 의한 주름 관련 유전자들의 발현량에 미치는 영향을 확인하기 위해 Hs27 세포의 RNA 추출 후, 실시간 정량 PCR(Real-time PCR)을 진행하였다.After extracting RNA from Hs27 cells to confirm the effect of the extract of each part by region (bud, leaf, root) on the expression level of wrinkle-related genes by UV irradiation in Hs27 cells, real-time quantitative PCR (Real-time PCR).
먼저 Hs27 세포를 배양하여 6 well plate에 2×105 cells/well 분주한 후 24시간 배양하였다. 배지를 제거한 후 DPBS로 세척하고 UVB lamp(5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan)를 이용하여 50 mJ/㎠조사한 후 양성대조군인 L-ascorbic acid 및 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)을 50, 100, 200 μg/mL 농도로 처리하여 24시간 배양하였다. 배양한 Hs27 세포의 RNA 추출은 RNeasy Mini kit (Qiagen, Valencia, CA, USA)를 이용하였고, iScript cDNA Synthesis kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA)로 cDNA를 합성하였다. First, Hs27 cells were cultured, 2 × 10 5 cells / well were dispensed into 6 well plates, and then cultured for 24 hours. After removing the medium, washed with DPBS, irradiated with 50 mJ / ㎠ using a UVB lamp (5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan), and positive control L-ascorbic acid and extracts by region (buds) , Leaves, roots) were cultured at 50, 100, 200 μg / mL concentration for 24 hours. RNA extraction of cultured Hs27 cells was performed using RNeasy Mini kit (Qiagen, Valencia, CA, USA), and cDNA was synthesized with iScript cDNA Synthesis kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
피부 주름 관련 유전자들의 발현을 측정하기 위하여 SYBR Green (iQ SYBR Green Supermix, Bio-Rad Laboratories Inc.)을 이용한 Real-Time PCR (Bio-Rad Laboratories, Inc., Hercules, CA, USA)을 이용하였다. 각각의 유전자에 대한 PCR primer의 염기서열은 하기 표 2에 나타내었다. Real-time PCR 반응은 총 20 μL 내에 10 μL의 2X SYBR mix, 2 μL의 cDNA, primer는 각각 100 pmol/μL를 1 μL씩 첨가하였고 나머지는 nuclease free water로 채워주었다. 모든 유전자에 대하여 PCR 증폭 단계는 다음과 같고 증폭 cycle은 40 cycle을 실시하였다. Hot start를 위해 95℃에서 10분, Denaturation을 95℃에서 15초, annealing을 58℃에서 30초, extension을 72℃에서 30초간 반복하며 각 cycle의 extension 후에 값이 기록되었다. 모든 cycle이 완료된 후 primer의 특이성을 확인하기 위해 melting curve 분석을 실시하였다. mRNA의 상대적인 함량은 자외선 처리를 하지 않은 군에 대한 상대적인 양으로 나타내었다.Real-Time PCR (Bio-Rad Laboratories, Inc., Hercules, CA, USA) using SYBR Green (iQ SYBR Green Supermix, Bio-Rad Laboratories Inc.) was used to measure the expression of skin wrinkle related genes. The base sequence of the PCR primers for each gene is shown in Table 2 below. In the real-time PCR reaction, 1 μL of 100 pmol / μL was added to each of 10 μL of 2X SYBR mix, 2 μL of cDNA, and primer in 20 μL, and the rest was filled with nuclease free water. PCR amplification steps for all genes were as follows, and the amplification cycle was 40 cycles. For the hot start, the value was recorded after the extension of each cycle by repeating for 10 minutes at 95 ° C, Denaturation at 95 ° C for 15 seconds, annealing at 58 ° C for 30 seconds, and extension at 72 ° C for 30 seconds. After all cycles were completed, melting curve analysis was performed to confirm the specificity of the primer. The relative content of mRNA was expressed as a relative amount to the group not treated with ultraviolet rays.
콜라겐(Collagen)은 진피의 85~90%를 차지하는 것으로 프로콜라겐(Procollagen)의 형태로 생성되었다가 효소 반응을 거쳐 피브릴(Fibril)을 형성하여 콜라겐 섬유를 형성하게 하는데, 콜라겐의 대부분을 차지하고 있는 것이 Collagen type I이다. Collagen accounts for 85 to 90% of the dermis. It is produced in the form of procollagen and then forms fibril through an enzymatic reaction to form collagen fibers, which occupy most of collagen. It is Collagen type I.
이와 관련된 유전자들의 발현을 확인한 결과, 도 4의 A)와 같이 Procollagen 유전자 발현량은 Normal control 군에 비해 control 군에서의 발현량이 유의적으로 감소하였다. 양하 추출물 처리군의 경우 control 군에 비해 농도 의존적으로 증가한 경향을 보이며 100, 200 μg/mL 농도에서 각각 39.8%, 52.3% 유의적 증가를 나타냈다. As a result of confirming the expression of related genes, the expression level of the Procollagen gene was significantly decreased in the control group compared to the normal control group, as shown in FIG. 4A). Yangha extract treated group showed a tendency to increase in a concentration-dependent manner compared to the control group, and showed a significant increase of 39.8% and 52.3% at 100 and 200 μg / mL concentration, respectively.
또한, 도 4의 (B)와 같이 Procollagen 유전자 발현량은 Normal control 군에 비해 control 군에서의 발현량이 유의적으로 감소하였다. 부위별 양하 추출물 처리군의 경우 control 군에 비해 Procollagen 유전자 발현량이 증가하였고(각 71%, 36%, 15%), 특히 양하 꽃봉오리 추출물(FSH-ZM)에서 유의적으로 가장 높은 발현 증가를 나타냈다. 또한, 도 4의 (C)와 같이 Collagen type I의 유전자 발현량은 Normal control군에 비해 control 군에서의 발현량이 유의적으로 감소하다. 양하 꽃봉오리 추출물 처리군의 경우 control군에 비해 농도 의존적으로 증가하는 경향을 보이며, 특히 200 μg/mL 농도에서 65.1% 유의적 증가를 나타냈다.In addition, as shown in FIG. 4 (B), the expression level of the Procollagen gene was significantly decreased in the control group compared to the normal control group. Compared to the control group, the expression group of yangha extract was increased by the expression level of procollagen gene (71%, 36%, 15%, respectively), and in particular, the expression of yangha bud extract (FSH-ZM) was the highest. . In addition, as shown in FIG. 4 (C), the expression level of the collagen type I is significantly reduced in the control group compared to the normal control group. The Yangha bud extract treatment group showed a tendency to increase in a concentration-dependent manner compared to the control group, and showed a significant increase of 65.1% at a concentration of 200 μg / mL.
한편, 자외선의 노출에 의해 증가하여 최종적으로 콜라겐을 가수분해하는데 관여하는 것으로 알려져 있는 MMPs(Matrix metalloproteinases) 유전자 발현량을 확인한 결과, 도 5와 같이 모든 MMPs에서 Normal control 군에 비해 control 군이 유의적으로 증가하였다. 이와 비교하여 양하 꽃봉오리 추출물 처리군의 경우 MMP1에 대해 양하 꽃봉오리 추출물 200 μg/mL에서 39.6% 유의적 감소를 나타냈고, MMP3에 대해 양하 꽃봉오리 추출물 100, 200 μg/mL 농도에서 각각 24.1%, 40.1% 유의적 감소를 나타냈으며, MMP9에 대해 양하 꽃봉오리 추출물 200 μg/mL에서 33% 유의적 감소를 나타냈다. Meanwhile, as a result of confirming the expression level of the matrix metalloproteinases (MMPs) known to be involved in hydrolysis of collagen by increasing by exposure to ultraviolet rays, as shown in FIG. 5, the control group was significantly higher than the normal control group in all MMPs. Increased to In comparison, the Yangha bud extract treatment group showed a significant decrease of 39.6% at 200 μg / mL of Yangha bud extract for MMP1, and 24.1% at concentrations of 100 and 200 μg / mL Yangha bud extract for MMP3, respectively. , 40.1%, and 33% significant reduction at 200 μg / mL of Yangha Bud Extract for MMP9.
또한, 자외선에 노출되면 염증반응이 촉진되게 되는데 염증반응을 일으키는 시토카인의 일종으로 면역 세포를 조절하는 TNF-α 유전자 발현량을 확인한 결과, 도 5와 같이 Normal control 군에 비해 control 군이 유의적으로 증가한 반면, 양하 꽃봉오리 추출물 처리군의 경우 control 군에 비해 양하 꽃봉오리 추출물 100, 200 μg/mL 농도에서 각각 28.8%, 43.7% 유의적 감소를 나타냈다.In addition, when exposed to UV rays, the inflammatory reaction is promoted. As a result of confirming the expression level of the TNF-α gene that regulates immune cells as a kind of cytokine that causes an inflammatory reaction, the control group is significantly higher than the normal control group as shown in FIG. 5. On the other hand, in the case of the Yangha bud extract treatment group, 28.8% and 43.7% of the
4) 양하 꽃봉오리 추출물이 염증성 사이토카인(Cytokine) 분비량에 미치는 영향 확인 실험4) Experiment to confirm the effect of Yangha bud extract on the amount of inflammatory cytokine secretion
상기 실시예의 양하 꽃봉오리 추출물이 Hs27 세포에서 자외선 조사에 의한 염증성 사이토카인 분비량에 미치는 영향을 확인하기 위해 하기와 같은 실험을 진행하였다. The following experiment was carried out to confirm the effect of the extract of yangha bud of the above example on the amount of inflammatory cytokine secretion by ultraviolet irradiation in Hs27 cells.
인간 섬유아세포인 Hs27 세포를 배양하여 96 well plate에 1×104 cells/well 분주한 후 24시간 배양하였다. 배지를 제거한 후 DPBS로 세척하고 UVB lamp(5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan)를 이용하여 50 mJ/㎠조사한 후 양성대조군인 L-ascorbic acid 및 양하 꽃봉오리 추출물을 50, 100, 200 μg/mL 농도로 처리하여 24시간 배양하였다. 염증성 사이토카인 IL-6, IL-1β를 측정하기 위해 Duoset ELISA kit (R&D system, Minneapolis, MN, USA)를 이용하여 검출하였으며 ELISA reader (Bio-Rad Laboratories Headquarters, Hercules, CA, USA)로 655 nm에서 흡광도를 측정하였다. 표준곡선(standard curve)를 이용하여 세포에서 생성된 사이토카인의 양을 계산하였다.Human fibroblast Hs27 cells were cultured, 1 × 10 4 cells / well were dispensed into 96 well plates, and cultured for 24 hours. After removing the medium, washed with DPBS, irradiated with 50 mJ / ㎠ using UVB lamp (5 Sankyo Denki G5T5 lamps, Sankyo Denki Co., Yokohama, Japan), and the positive control L-ascorbic acid and
자외선의 노출은 피부에서 활성산소종의 생성을 자극하여 세포막 지질의 산화적 손상을 유발함으로써 염증성 사이토카인의 분비를 촉진시키게 된다. 염증성 사이토카인의 일종인 IL-1β의 분비량을 확인한 결과, 도 6과 같이 Normal control군에 비해 control 군이 유의적으로 증가한 반면, 양하 꽃봉오리 추출물 처리군의 경우 control 군에 비해 농도 의존적으로 감소하는 경향을 보이며 특히 200 μg/mL농도에서만 27.4% 유의성 감소를 나타냈다. 또한, 염증성 사이토카인의 또 다른 일종인 IL-6의 분비량을 확인한 결과, Normal control군에 비해 control 군이 유의적으로 증가한 반면, 양하 꽃봉오리 추출물 처리군의 경우 control 군에 비해 200 μg/mL농도에서 18.2% 유의성 감소를 나타냈다.Exposure to UV rays stimulates the production of free radicals in the skin, causing oxidative damage to cell membrane lipids, thereby promoting the secretion of inflammatory cytokines. As a result of confirming the secretion amount of IL-1β, a type of inflammatory cytokine, as shown in FIG. 6, the control group was significantly increased compared to the normal control group, whereas the yangha bud extract treatment group was concentration-dependently decreased compared to the control group. It showed a tendency to show a 27.4% decrease in significance, especially at the concentration of 200 μg / mL. In addition, as a result of confirming the secretion amount of IL-6, another type of inflammatory cytokine, the control group significantly increased compared to the normal control group, whereas in the case of the yangha bud extract treatment group, the concentration of 200 μg / mL compared to the control group The 18.2% decrease in significance was observed.
이와 같이 인간섬유아 세포인 Hs27 세포에서 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)은 control 군에 비해 콜라겐 관련 유전자 Procollagen의 발현량을 증가시켰고, 특히 양하 꽃봉오리 추출물은 다른 부위별 양하 추출물(잎, 뿌리)에 비해 Procollagen의 발현량을 크게 증가시켰다. 또한, 양하 꽃봉오리 추출물은 control 군에 비해 콜라겐 관련 유전자 Collagen type I의 발현량을 증가시켰고, 콜라겐 가수분해 관련 유전자 MMPs와 염증성 사이토카인 TNF-α의 발현량을 감소시켰고, 염증성 사이토카인인 IL-1β 및 IL-6의 분비량을 감소시켰다. 이를 통해 자외선 노출에 의해 콜라겐의 가수분해에 관여하는 MMPs 관련 기전에 있어서 양하 꽃봉오리가 억제 효과를 나타냄으로써 피부 주름 개선 효능을 나타내는 물질로 이용할 수 있음을 알 수 있다.Thus, in human fibroblasts, Hs27 cells, extracts by region (buds, leaves, and roots) increased the expression level of the collagen-related gene Procollagen compared to the control group. Leaf, root) compared to Procollagen expression significantly increased. In addition, Yangha bud extract increased the expression level of collagen-related gene Collagen type I compared to the control group, decreased the expression level of collagen hydrolysis-related gene MMPs and inflammatory cytokine TNF-α, and the inflammatory cytokine IL- The secretion of 1β and IL-6 was reduced. Through this, it can be seen that, in the mechanism related to MMPs involved in the hydrolysis of collagen by exposure to ultraviolet rays, the yangha bud can be used as a substance showing the effect of improving skin wrinkles by exhibiting an inhibitory effect.
[실험예 3 : 양하 추출물의 피부 미백 효과 확인][Experimental Example 3: Confirmation of skin whitening effect of Yangha extract]
본 실험예에서는 상기 실시예에서 제조한 양하 추출물의 피부 미백에 대한 효과를 확인하고자 하였다.In this experimental example, it was intended to confirm the effect on the skin whitening of the extract of yangha prepared in the above example.
1) 세포 배양 1) Cell culture
실험에 사용된 멜라닌생성세포인 B16F10은 American Type Cultured Collection (ATCC; Rockville, MD, USA)에서 분주 받아 실험하였다. 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (100 units/mL)이 함유된 High-glucose Dulbeco’s modified Eagle’s medium배지를 사용하였고, 37℃, 5% CO₂, 95% humid air로 조절된 배양기(Thermo Fisher Scientific Inc., Pittsburgh, PA, USA)에서 배양하였다.The melanocytes used in the experiment, B16F10, were tested after being dispensed from the American Type Cultured Collection (ATCC; Rockville, MD, USA). High-glucose Dulbeco's modified Eagle's medium medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (100 units / mL) was used, and incubator adjusted to 37 ℃, 5% CO₂, 95% humid air (Thermo Fisher Scientific Inc., Pittsburgh, PA, USA).
2) 세포 독성 시험2) Cytotoxicity test
B16F10 세포를 96-well plate에 1×104 cells/well로 분주하여 24시간 동안 배양한 후, 각 well에 양하 꽃봉오리 추출물을 농도별로 처리하였다. 24시간 후, 5 mg/ml 농도의 MTT solution을 20 μL 첨가하고 4시간 동안 배양한 후, 배지를 제거하고 DMSO 100 μL/well 씩 첨가하여 560 nm에서 흡광도를 측정하였다.After dispensing B16F10 cells in a 96-well plate at 1 × 10 4 cells / well and incubating for 24 hours, each well was treated with extracts of yangha buds by concentration. After 24 hours, after adding 20 μL of 5 mg / ml MTT solution and incubating for 4 hours, the medium was removed and DMSO was added at 100 μL / well to measure absorbance at 560 nm.
그 결과, 도 1의 (C)와 같이 양하 꽃봉오리 추출물 200 μg/mL이하의 농도에서 세포독성이 나타나지 않는 것을 확인하여 본 실험에서는 양하 꽃봉오리 추출물 200 μg/mL농도까지 처리하였다.As a result, as shown in (C) of FIG. 1, it was confirmed that cytotoxicity was not observed at a concentration of 200 μg / mL or less of the yangha bud extract, and in this experiment, the yangha bud extract was treated to a concentration of 200 μg / mL.
3) 양하 추출물의 멜라닌 생성 효소 활성 억제 확인 실험3) Experiment to confirm the inhibition of melanin-producing enzyme activity of Yangha extract
상기 실시예의 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)이 멜라닌 생성세포인 B16F10 세포에서 멜라닌 반응을 조절하는 대표적인 효소 티로시나제(tyrosinase)의 활성에 미치는 영향을 확인하기 위해 하기와 같이 실험을 진행하였다.The experiments were conducted as follows to confirm the effect of the extract of each part by region (bud, leaf, root) on the activity of tyrosinase, a representative enzyme that regulates the melanin response in B16F10 cells, which are melanocytes. .
먼저 B16F10 세포를 배양하여 24 well plate에 5×104cells/well에 분주하고 24시간 동안 배양한 후, 각 well에 양성대조군인 알부틴(arbutin)과 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)을 50, 100, 200 μg/mL 농도로 처리함과 동시에 분화시약인 IBMX 250 μM을 처리하여 3일간 멜라닌 색소의 분화를 유도하였고 24시간마다 추출물과 배지를 교체하였다. 72시간 배양한 후 각 well을 DPBS로 세척하고 Cell lysis reagent buffer 125 μL를 분주하여 세포를 분리하였다. 상층액을 분리하여 Bradford Dye Reagent로 단백질을 정량하고, 세포에 1N NaOH에 10% DMSO가 함유된 용액을 300 μL를 첨가하여 100 ℃에서 10분 동안 용해시켜 ELISA reader(Bio-Rad Laboratories Headquarters, Hercules, CA, USA)로 450 nm에서 흡광도를 측정하였다.First, B16F10 cells were cultured, dispensed into 5 × 10 4 cells / well in a 24 well plate, and incubated for 24 hours, followed by positive control of arbutin in each well and extracts from each region (buds, leaves, and roots). Was treated with concentrations of 50, 100, and 200 μg / mL, and at the same time, treatment with 250 μM of the differentiation reagent, induced differentiation of melanin pigment for 3 days, and extract and medium were replaced every 24 hours. After incubation for 72 hours, each well was washed with DPBS and 125 μL of Cell lysis reagent buffer was dispensed to separate cells. The supernatant was separated to quantify proteins with Bradford Dye Reagent, and 300 μL of a solution containing 10% DMSO in 1N NaOH was added to cells to dissolve for 10 minutes at 100 ° C. and ELISA reader (Bio-Rad Laboratories Headquarters, Hercules) , CA, USA) was measured at 450 nm.
티로시나제(Tyrosinase)는 멜라닌 생성의 첫 단계에 관여하는 효소로 티로신을 히드록시화하여 디하이드록시페닐알라닌(dihydroxyphenylalanine, DOPA)을 형성하고, 더 산화되어 도파퀴논을 형성하는 역할을 한다. 멜라닌(Melanine)은 색소 중 하나로써 흑갈색 또는 검은색을 띠며 멜라닌의 양에 따라 색의 짙은 정도가 결정된다.Tyrosinase (Tyrosinase) is an enzyme involved in the first step of melanin production, which hydroxylates tyrosine to form dihydroxyphenylalanine (DOPA), and is further oxidized to form dopaquinone. Melanine (Melanine) is one of the pigments, dark brown or black, and the color of melanin is determined depending on the amount of melanin.
양하 추출물이 티로시나제 활성에 미치는 영향을 확인한 결과, 도 7의 A)와 같이 양하 꽃봉오리 추출물 처리군의 경우 control 군에 비해 50, 100, 200 μg/mL 농도에서 각각 7.3%, 13.2%, 18.3%로 농도 의존적으로 유의성 감소를 나타냈다. 또한, 도 7의 B)와 같이 부위별 양하 추출물 처리군의 경우 control 군에 비해 티로시나제 활성이 감소하였고(각 14%, 10%, 2%), 특히 양하 꽃봉오리 추출(FSH-ZM)에서 티로시나제 활성에 가장 유의적으로 높은 감소를 나타냈다.As a result of confirming the effect of yangha extract on tyrosinase activity, in the case of the yangha bud extract treatment group as shown in FIG. 7A), 7.3%, 13.2%, and 18.3% at 50, 100, and 200 μg / mL concentrations, respectively, compared to the control group. Significantly decreased in concentration-dependent manner. In addition, as shown in B) of FIG. 7, the tyrosinase activity was decreased in the case of the yangha extract extract by region compared to the control group (14%, 10%, 2%, respectively), especially in the yangha bud extract (FSH-ZM). It showed the most significant decrease in activity.
4) 양하 꽃봉오리 추출물의 멜라닌 합성 억제능 확인 실험4) Experiment to confirm the ability to inhibit melanin synthesis of Yangha bud extract
상기 실시예의 양하 꽃봉오리 추출물이 멜라닌 생성세포인 B16F10 세포에 대한 멜라닌 합성 억제능을 확인하기 위해 하기와 같이 실험을 진행하였다.The experiment was conducted as follows to confirm the melanin synthesis inhibitory ability of the yangha bud extract of the above Example for B16F10 cells, which are melanocytes.
멜라닌 합성에 관여하는 티로시나제를 측정하기 위해 양성대조군인 알부틴(arbutin)과 양하 꽃봉오리 추출물을 용매에 녹이고, 0.1M 인산염완충액(pH 6.5) 140 μL, 시료액 100 μL, 1.5 mM L-DOPA(L-3,4- dihydroxy-phenylalanine) 40 μL, mushroom tyrosinase(2000 U/mL) 20 μL를 순서대로 넣었다. 37 ℃ 10분 동안 반응시킨 다음 490 nm에서 흡광도를 측정하였다.In order to measure tyrosinase involved in melanin synthesis, the positive control arbutin and the extract of the bud bud were dissolved in a solvent, 140 μL of 0.1M phosphate buffer (pH 6.5), 100 μL of sample solution, and 1.5 mM L-DOPA (
멜라닌 생성세포인 B16F10 세포는 세포 내 cAMP가 증가되면 MITF의 발현을 증가시켜 멜라닌 합성을 유도하는데 본 실험에서는 cAMP를 자극하는 유도제로 IBMX를 사용하였고 양하 꽃봉오리 추출물을 함께 처리하면서 멜라닌 생성 억제능을 확인하였다. 그 결과, 도 8과 같이 Normal control 군에 비해 IBMX로 분화를 유도한 control 군의 멜라닌 농도(melanin contents)가 유의적 증가를 나타냈다. 이에 비교하여 양하 꽃봉오리 추출물 처리군의 경우, control 군에 비해 50, 100, 200 μg/mL에서 각각 47.6%, 51%, 54.8%로 농도 의존적으로 유의성 감소를 나타냈다.B16F10 cells, which are melanin-producing cells, increase the expression of MITF when intracellular cAMP is increased to induce melanin synthesis. In this experiment, IBMX was used as an inducer to stimulate cAMP, and the melanin production inhibitory ability was confirmed while processing the extracts of yangha bud together. Did. As a result, as shown in FIG. 8, the melanin contents of the control group induced differentiation with IBMX showed a significant increase compared to the normal control group. Compared to this, in the case of the Yangha bud extract treatment group, it showed a concentration-dependent decrease in concentration at 47.6%, 51%, and 54.8% at 50, 100, and 200 μg / mL, respectively, compared to the control group.
이와 같이 멜라닌 생성세포인 B16F10 세포에서 부위별 양하 추출물(꽃봉오리, 잎, 뿌리)은 멜라닌 생성에 관여하는 효소 티로시나제 활성을 억제시켰고, 특히, 양하 꽃봉오리 추출물은 다른 부위별 양하 추출물(잎, 뿌리)에 비해 가장 높은 억제 효과를 나타냈다. 또한, 양하 꽃봉오리 추출물은 control 군에 비해 멜라닌 생성량을 감소키는 것으로 나타났다. 이를 통해 양하 꽃봉오리는 흑갈색 또는 검은색 색소인 멜라닌 생성 기전에 있어서 억제 효과를 나타냄으로써 피부 미백 효능을 나타내는 물질로 이용할 수 있음을 알 수 있다.As described above, in the B16F10 cells, which are melanin-producing cells, the extracts by region (buds, leaves, and roots) inhibited the activity of the enzyme tyrosinase involved in melanin production. ). In addition, it was found that Yangha bud extract reduced melanin production compared to the control group. Through this, it can be seen that the Yangha flower bud can be used as a substance showing skin whitening efficacy by exhibiting an inhibitory effect in the mechanism of melanin production, which is a black-brown or black pigment.
이상과 같이, 본 발명의 식품 또는 화장료 조성물은 양하 추출물을 함유함에 따라 피부 보습 효과, 피부 주름 개선 효과 및 피부 미백 효과가 우수하고, 특히 양하 꽃봉오리 추출물 고농도(200 μg/mL)에서 공통적으로 그 효능이 가장 뛰어나며, 양성대조군의 50, 100 μg/mL과 유사한 결과를 나타내는 것을 확인하였다. As described above, the food or cosmetic composition of the present invention is excellent in skin moisturizing effect, skin wrinkle improvement effect and skin whitening effect as it contains a Yangha extract, especially in a high concentration (200 μg / mL) of Yangha bud extract. It was confirmed that the efficacy was the best, and the results showed similar results to 50 and 100 μg / mL of the positive control.
이에 본원발명의 양하(특히, 양하 꽃봉오리) 추출물을 이용한 식품 또는 화장료 조성물은 부작용 위험이 낮으며 피부 보습을 통해 피부 장벽을 견고하게 하고, 피부 주름과 멜라닌 색소의 형성을 억제하여 효과적인 피부 건강 기능성을 가진다. Thus, the food or cosmetic composition using Yangha (especially, Yangha bud) extract of the present invention has a low risk of side effects and strengthens the skin barrier through moisturizing the skin, and suppresses the formation of skin wrinkles and melanin pigments, thereby effective skin health function Have
Claims (10)
Yangha ( Zingiber mioga ) Food composition for skin moisturizing, skin wrinkle improvement and skin whitening, characterized by containing a bud extract as an active ingredient.
After adding 1200 to 1800 parts by weight of purified water to 100 to 400 parts by weight of Yangha ( Zingiber mioga ) bud, extracting it for 6 to 10 hours at 80 to 90 ° C to obtain a Yangha bud extract. A method for preparing a food composition for moisturizing skin, improving skin wrinkles, and whitening skin.
상기 양하 꽃봉오리 추출물은,
양하 꽃봉오리 열수 추출물인 것을 특징으로 하는 식품 조성물.
According to claim 1,
The Yangha bud extract,
Food composition, characterized in that the Yangha bud hot water extract.
Yangha ( Zingiber mioga ) Cosmetic composition for moisturizing skin, improving skin wrinkles and whitening skin, characterized by containing a bud extract as an active ingredient.
After adding 1200 to 1800 parts by weight of purified water to 100 to 400 parts by weight of Yangha ( Zingiber mioga ) bud, extracting it for 6 to 10 hours at 80 to 90 ° C to obtain a Yangha bud extract. A method for preparing a cosmetic composition for moisturizing skin, improving skin wrinkles, and whitening skin.
상기 양하 꽃봉오리 추출물은,
양하 꽃봉오리 열수 추출물을 특징으로 하는 화장료 조성물.The method of claim 6,
The Yangha bud extract,
Cosmetic composition characterized by Yangha bud hot water extract.
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| KR20110001538A (en) * | 2009-06-30 | 2011-01-06 | 한국콜마 주식회사 | Flowers extract having anti-oxidation and whitening effects and extraction method thereof and cosmetics comprising flowers extract |
| KR20110032715A (en) * | 2009-09-24 | 2011-03-30 | 두리화장품 주식회사 | Antiaging cosmetic compositions and method for producing thereof |
| KR20140059958A (en) * | 2012-11-09 | 2014-05-19 | 주식회사 유니베라 | Composition for obesity treatment and improvement which contains extract of flower petals of zingiber mioga |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20110001538A (en) * | 2009-06-30 | 2011-01-06 | 한국콜마 주식회사 | Flowers extract having anti-oxidation and whitening effects and extraction method thereof and cosmetics comprising flowers extract |
| KR20110032715A (en) * | 2009-09-24 | 2011-03-30 | 두리화장품 주식회사 | Antiaging cosmetic compositions and method for producing thereof |
| KR20140059958A (en) * | 2012-11-09 | 2014-05-19 | 주식회사 유니베라 | Composition for obesity treatment and improvement which contains extract of flower petals of zingiber mioga |
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