KR20140103439A - Composition for preventing, improving or treating allergic dermatitis comprising hot water extract or ferment extract of chestnut inner shell - Google Patents
Composition for preventing, improving or treating allergic dermatitis comprising hot water extract or ferment extract of chestnut inner shell Download PDFInfo
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- KR20140103439A KR20140103439A KR1020130016792A KR20130016792A KR20140103439A KR 20140103439 A KR20140103439 A KR 20140103439A KR 1020130016792 A KR1020130016792 A KR 1020130016792A KR 20130016792 A KR20130016792 A KR 20130016792A KR 20140103439 A KR20140103439 A KR 20140103439A
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- Prior art keywords
- extract
- present
- fermented
- hot water
- skin
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Abstract
Description
본 발명은 율피 열수추출물 또는 율피 발효추출물을 함유하는 알레르기성 피부질환 예방, 개선 또는 치료용 조성물에 관한 것이다. The present invention relates to a composition for preventing, ameliorating or treating an allergic skin disease containing a watery urine extract or a uricellium fermented extract.
산업화·도시화의 심화로 식생활과 주거생활의 변화, 환경오염에 의한 화학적·생물학적 유해물질에 대한 노출로 다양한 연령층에서 알레르기 질환이 크게 증가하고 있는 실정이다. 이는 인체가 무해한 외부 환경에 대해서도 민감한 반응을 보이며, 천식이나 비염과 같은 호흡기 질환 또는 접촉성 피부염, 아토피 질환 등의 피부 질환 등으로 나타나게 된다. Allergic diseases are increasing in various age groups due to changes in diet and living conditions, exposure to chemical and biological harmful substances due to environmental pollution, and so on. This is a sensitive response to the harmless external environment, and is caused by respiratory diseases such as asthma and rhinitis, or skin diseases such as contact dermatitis and atopic diseases.
특히 피부는 외부 알레르기 유발 물질이 공기, 음식 등에 의해 직접적으로 접촉되어 과민한 면역 반응을 일으키게 된다. 알레르기성 피부 질환은 음식물, 세균, 진드기, 기후환경인자 등 외부 항원에 지속적으로 노출되어 발병하는 염증 질환으로 개인의 유전적인 특성 때문에 다양한 환경 인자들에 대한 알레르기 반응이 개개인마다 다르게 나타난다. 이들 질환을 개선, 치료하기 위해서는 우선 알레르기 유발 요인에 노출 및 접촉을 피하고 피부를 청결하게 유지하면서 적절한 약효성분을 피부염 발병 부위에 도포하거나 전신에 투여하는 방법이 있다. In particular, the skin is directly contacted with external allergens by air, food, etc., and causes an immune immune response. Allergic skin diseases are inflammatory diseases that are continuously exposed to external antigens such as food, bacteria, mites, and climate environmental factors. Individual allergic reactions to various environmental factors are different because of their genetic characteristics. In order to improve or treat these diseases, there is a method in which exposure to the allergen causing factors and contact are avoided and the skin is kept clean, and the appropriate medicinal active ingredient is applied to the site of the dermatitis or administered to the whole body.
그러나 합성 알레르기성 피부염 억제제는 가격이 고가이고, 의약품 및 화장품적인 방법 모두 아직까지는 병변의 재발을 막을 수 없어 외부 항원 노출 시 재발이 빈번하게 일어나며 부작용이 검증되지 않는 등의 문제점을 지니고 있어 이를 보완하기 위해 천연소재를 이용한 치료제 개발 필요성이 절실한 실정이다. However, synthetic allergenic dermatitis inhibitors are expensive, and drug and cosmetic methods can not prevent the recurrence of lesions. Therefore, recurrence frequently occurs when external antigens are exposed, and side effects are not proved. The need for the development of therapeutic agents using natural materials is urgent.
최근 천연물로부터 새로운 물질에 대한 탐색 및 연구와 이에 따른 유효성·안전성 평가 기준이 확립되면서 천연물을 이용한 약물개발 연구가 가속화되고 있으며, 알레르기성 피부염 개선의 일환으로 한약재를 이용하거나 황벽나무, 대나무, 다래, 버섯 등 식물이나 수목의 추출물을 단독 또는 혼합하여 이용한 치료제의 개발이 활발하게 진행되고 있다.Recently, researches and researches on new substances from natural products have been established, and researches on the development of drugs using natural materials have been accelerated. As a result, Mushrooms, and the like, are being actively developed.
우리나라는 예로부터 김치, 된장, 간장 등과 같은 발효식품을 많이 섭취하고 있으며, 최근 발효식품의 생리활성 작용이 알려지면서 건강 기능성 소재로 응용하기 위한 연구가 이루어지고 있다. 발효는 넓은 뜻으로 미생물에 의한 유용한 물질 생산으로 여길 수 있는데, 미생물은 영양물질을 외부로부터 세포 내로 섭취하고, 이들 영양물질이 효소에 의해 분해되어 생성되는 에너지를 이용하여 생명의 유지와 생장이 가능하도록 해준다. 최근 캘리포니아 대학의 연구팀의 결과에 따르면, 프로바이오틱 성분이 포함된 보충제를 복용한 아이들에게 알레르기성 피부 질환을 예방, 억제 효과를 보이는 것으로 나타났으며 이들 성분 중에서도 Lactobacillus rhamnosus GG가 장기적으로 예방 효과가 큰 것으로 나타났다고 보고되었다. Korea has been consuming a lot of fermented foods such as kimchi, soybean paste, soy sauce and so on. Since the physiological activity of fermented foods has recently been known, studies for application as health functional materials have been made. Fermentation can be regarded as a wide range of usable materials production by microorganisms. Microorganisms can take the nutrients from the outside into the cells and maintain and grow life by using the energy generated by decomposition of these nutrients by enzymes. It will. Recently, researchers at the University of California have shown that children who take a supplement containing probiotics can prevent and inhibit allergic skin disease. Among these ingredients, Lactobacillus rhamnosus GG has long-term preventive effects , Respectively.
프로바이오틱은 대장에서 항생물질의 작용을 돕는 몸에 유익한 박테리아를 아우르는 단어로 발효식품에 다량 함유되어 있는 성분이며, 항알레르기성을 지닌 레몬밤, 도라지, 인삼, 알로에 등 다양한 식품소재를 발효 또는 첨가하여 식품의 기능성 강화와 더불어 품질을 향상시킨 발효식품으로서 개발하려는 시도가 이루어지고 있다.The term "probiotic" refers to bacteria that are beneficial to the body to help antibiotics in the large intestine. It is a substance that is contained in fermented foods. It is a fermented food that contains various anti-allergic ingredients such as lemon balm, bellflower, ginseng, An attempt has been made to develop a fermented food having improved quality as well as enhanced functionality of the food.
밤나무(Castanea crenata Sieb)는 참나무과에 속하는 낙엽 활엽 교목으로 그 열매인 밤(Castane crenata)은 식용, 약용, 가공식품으로 이용되고 있으며, 열매뿐 아니라 생잎, 밤나무껍질, 밤 열매 껍질과 내피 등도 약재로 사용되고 있다. Chestnut (Castanea crenata Sieb) is a deciduous broad-leaved arboreous tree belonging to the oak family. Castane crenata is used for edible, medicinal and processed foods. It is also used as a medicinal material such as raw leaves, chestnut husks, chestnut husks and endothelium. .
율피를 이용한 종래의 기술로는 대한민국 공개특허 제10-2010-0088007호에 율피 에탄올 추출물을 포함하는 지방간의 예방 또는 치료용 조성물에 관하여 개시하고 있으나 율피 발효추출물의 제조방법 및 항알러지 또는 항아토피 효과에 대하여는 전혀 교시 또는 암시된 바 없다.Korean Patent Laid-Open No. 10-2010-0088007 discloses a composition for prevention or treatment of fatty liver comprising a uri-fructose ethanol extract. However, the present invention relates to a method for producing a fermented extract and a method for producing an anti-allergic or anti- Have not been taught or implied at all.
따라서 본 발명의 목적은 율피 열수추출물 또는 율피 발효추출물을 함유하는 알레르기성 피부질환 예방 및 치료용 약학적 조성물을 제공하는 데 있다.Accordingly, an object of the present invention is to provide a pharmaceutical composition for prevention and treatment of allergic skin diseases containing a hydrolyzed extract of U. P. or a U. fermented extract.
본 발명의 상기 목적은 율피 열수추출물을 제조하는 단계와: 상기 단계에서 수득한 율피 열수추출물을 유산균을 통하여 발효시켜 율피 발효추출물을 수득하는 단계와; 상기 단계들에서 수득한 율피 열수추출물 및 율피 발효추출물의 총 폴리페놀 및 총 플라보노이드 함량 분석과 항산화, 항염 및 항아토피 효과를 각각 평가하는 단계를 통하여 달성하였다.The above object of the present invention can be achieved by a method for producing a fermented hot-water extract, comprising the steps of: preparing a hot-water extract of Uru, and fermenting the hot-water extract of Uru- The total polyphenol and total flavonoid content and the antioxidant, anti-inflammatory and anti-atopic effects of the UFI hot water extract and the UFI fermentation extract obtained in the above steps were evaluated through the respective steps.
본 발명은 율피 열수추출물 또는 율피 발효추출물을 함유하는 알레르기성 피부질환 예방, 개선 또는 치료용 조성물을 제공하는 뛰어난 효과가 있다.The present invention has an excellent effect of providing a composition for preventing, ameliorating or treating an allergic skin disease containing a watery uric extract or a uricellium fermented extract.
도 1은 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 총 폴리페놀 함량을 측정한 실험결과를 나타낸 그래프이다.
도 2는 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 총 플라보노이드 함량을 측정한 실험결과를 나타낸 그래프이다.
도 3은 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 전자공여능을 통하여 항산화 활성을 측정한 실험결과를 나타낸 그래프이다.
도 4는 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 superoxide anion radical 소거능을 통하여 항산화 활성을 측정한 실험결과를 나타낸 그래프이다.
도 5는 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 세포독성을 측정한 실험결과를 나타낸 그래프이다.
도 6은 본 발명 율피 열수추출물 및 율피 발효추출물 각각의 항염 활성을 측정한 실험결과를 나타낸 그래프이다.
도 7은 본 발명 율피 열수추출물 및 율피 발효추출물 각각을 아토피유발 마우스에 처리한 다음 알레르기치료 효과를 촬영한 사진도이다.
도 8은 본 발명 율피 열수추출물 또는 율피 발효추출물 각각을 아토피유발 마우스에 처리한 다음 마우스 피부조직으로부터 피부멜라닌지수(A), 피부홍반지수(B) 및 피부수분지수(C)를 측정한 결과를 나타낸 그래프이다.
도 9는 본 발명 율피 열수추출물 또는 율피 발효추출물 각각을 아토피유발 마우스에 처리한 다음 마우스 피부조직의 염증유발 유전자들의 발현분석결과를 나타낸 그래프이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the results of an experiment in which the total polyphenol content of each of the watery extract of the present invention and the fermented extract of Pui is measured.
FIG. 2 is a graph showing the results of an experiment in which the total flavonoid content of each of the watery fruit extract of the present invention and the fermented extract of Julia pygmae was measured.
FIG. 3 is a graph showing the results of an experiment in which the antioxidative activity was measured by the electron donating ability of each of the watery fruit extract of the present invention and the uri fermented extract.
FIG. 4 is a graph showing the results of an experiment in which antioxidative activity was measured by superoxide anion radical scavenging activity of each of the watery extract of U.
FIG. 5 is a graph showing the results of an experiment for measuring cytotoxicity of each of the watery fruit extract of the present invention and the fermented extract of Ufu.
FIG. 6 is a graph showing the results of an experiment for measuring the anti-inflammatory activity of each of the watery extract of the present invention and the fermented extract of Julia.
FIG. 7 is a photograph showing the effect of treatment of allergy with treating the atopy-induced mouse of the present invention with the watery extract of the present invention and the yeast extract, respectively.
8 is a graph showing the results of measuring the skin melanin index (A), skin eruption index (B) and skin moisture index (C) from mouse skin tissues after treating each of the hot water or hot water extracts of the present invention with atopy- Fig.
FIG. 9 is a graph showing the results of analysis of the expression of inflammatory genes in mouse skin tissues after treating each of the hot-water extract of U. P. or the U. fermented extract of the present invention with atopy-induced mice.
본 발명에 있어서 알러지성 피부질환은 접촉성 피부염, 아토피 질환 등의 피부 질환이다.In the present invention, allergic skin diseases are skin diseases such as contact dermatitis and atopic diseases.
본 발명은 본 발명에 따라 제조된 율피 열수추출물 및 율피 발효추출물을 유효성분으로 함유하는 약학적 조성물을 제공한다. 상기 율피 열수추출물 또는 율피 발효추출물은 바람직하게 약학적 조성물 총 중량에 대하여 0.1 내지 99 중량%를 함유하는 것으로 한다.The present invention provides a pharmaceutical composition containing as an active ingredient an udder extract of hot pepper and a fermented extract of Aspergillus sieboldii according to the present invention. The watery extract or watery extract of UFI is preferably contained in an amount of 0.1 to 99% by weight based on the total weight of the pharmaceutical composition.
본 발명에 따른 상기 약학적 조성물은 약학적으로 허용 가능한 담체를 포함하여 다양한 형태, 예를 들어 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구용 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제제화될 수 있다. The pharmaceutical composition according to the present invention may be formulated into various forms including pharmaceutically acceptable carriers, for example oral preparations, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, Can be formulated in the form of sterile injectable solutions.
상기 약제학적으로 허용 가능한 담체는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유 등을 포함한다. 또한, 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 포함한다. 경구용 고형 제제는 정제, 환제, 산제, 과립제, 캡슐제 등을 포함하며, 이러한 고형제제는 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트 (calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 포함할 수 있으며, 마그네슘 스테아레이트, 탈크 같은 윤활제 등을 포함할 수 있다. 경구용 액상 제제는 현탁제, 내용액제, 유제, 시럽제 등을 포함하며, 물, 리퀴드 파라핀 등의 희석제, 습윤제, 감미제, 방향제, 보존제 등을 포함할 수 있다. 비경구용 제제는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제를 포함하며, 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르류 등을 포함한다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈 (tween) 61, 카카오지, 라우린지, 글리세로 젤라틴 등이 사용될 수 있다.
The pharmaceutically acceptable carrier may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. It also includes diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Solid form preparations for oral use include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose ), Gelatin and the like, and may include a lubricant such as magnesium stearate, talc, and the like. Oral liquid preparations include suspensions, solutions, emulsions, syrups, and the like, and may contain diluents such as water and liquid paraffin, wetting agents, sweetening agents, fragrances, preservatives and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, ethylcellulose, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, polyvinylpyrrolidone, And the like, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
본 발명은 본 발명에 따라 제조된 율피 열수추출물 또는 율피 발효추출물을 유효성분으로 함유하는 건강기능성식품 조성물을 제공한다. 상기 율피 열수추출물 또는 율피 발효추출물은 바람직하게 건강기능성식품 조성물 총 중량에 대하여 0.1 내지 99 중량%를 함유하는 것으로 한다.The present invention provides a health functional food composition comprising as an active ingredient a tubular hot-water extract or a tuberous fermented extract prepared according to the present invention. Preferably, the watery extract or the fermented extract of UFI contains 0.1 to 99% by weight based on the total weight of the health functional food composition.
본 발명의 식품 조성물은 건강기능식품으로서 사용될 수 있다. 상기 "건강기능식품"이라 함은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The food composition of the present invention can be used as a health functional food. The term "functional food" as used herein means a food prepared and processed using a raw material or ingredient having a useful function in the human body pursuant to Law No. 6727 on Health Functional Foods, and the term "functional" And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.
본 발명의 식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 "식품 첨가물"로 서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전청에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The food composition of the present invention may contain conventional food additives, and the suitability of the above-mentioned "food additives" is not limited by the general rules and general test methods approved by the Food and Drug Administration It shall be determined according to standards and standards for the item.
상기 "식품 첨가물 공전"에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류들을 들 수 있다.Examples of the substances found in the above-mentioned "food additives" include natural compounds such as ketones, chemical compounds such as glycine, potassium citrate, nicotinic acid and cinnamic acid, coloring pigments, licorice extracts, crystalline cellulose, high- L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar pigment preparations and the like.
캅셀 형태의 건강기능식품 중 경질 캅셀제는 통상의 경질캅셀에 생약 추출물에 부형제 등의 첨가제와의 혼합물 또는 그의 입상물 또는 제피한 입상물을 충진하여 제조할 수 있으며, 연질 캅셀제는 생약 추출물에 부형제 등의 첨가제와의 혼합물을 젤라틴 등 캅셀기제에 충진하여 제조할 수 있다. 상기 연질 캅셀제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.The hard capsule of the capsule type health functional food can be prepared by filling a normal hard capsule with a mixture of herbal medicine extract with an additive such as an excipient or a granular product thereof or a granulated product with a hard gelatinized product. Of the present invention may be filled in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.
환 형태의 건강기능식품은 생약 추출물에 부형제, 결합제, 붕해제 등의 혼합물을 적당한 방법으로 성형하여 조제할 수 있으며, 필요에 따라 백당이나 다른 적당한 제피제로 제피를, 또는 전분, 탈크 또는 적당한 물질로 환의를 입힐 수도 있다.In case of a ring type health functional food, a mixture of excipient, binder, disintegrant, etc. may be formulated into a herbal medicine extract by an appropriate method. If necessary, the preparation may be mixed with white sugar or other suitable skin care agent or a starch, talc or a suitable substance It is possible to make a ring.
과립형태의 건강기능식품은 상기 생약 추출물에 부형제, 결합제, 붕해제 등의 혼합물을 적당한 방법으로 입상으로 제조할 수 있으며, 필요에 따라 착향제, 교미제 등을 함유할 수 있다. 본원 발명의 상기 부형제, 결합제, 붕해제, 활택제, 교미제, 착향제 등에 대한 용어 정의는 당업계에 공지된 문헌에 기재된 것으로 그 기능 등이 동일 내지 유사한 것들을 포함한다 (대한약전 해설편, 문성사, 한국약학대학협의회, 제 5 개정판, p33-48, 1989).In the granular health functional food, a mixture of an excipient, a binder and a disintegrant may be prepared into a granular form by an appropriate method, and if necessary, a flavoring agent, a mating agent and the like may be contained in the herbal medicine extract. The definitions of the above-mentioned excipients, binders, disintegrants, lubricants, mating agents, flavoring agents and the like of the present invention are described in documents known in the art and include the same or similar functions (see, for example, Korean Association of Pharmacy, 5th ed., Pp. 33-48, 1989).
본 발명은 본 발명에 따라 제조된 율피 열수추출물 또는 율피 발효추출물을 유효성분으로 함유하는 화장료 조성물을 제공한다. 상기 율피 열수추출물 또는 율피 발효추출물은 바람직하게 화장료 조성물 총 중량에 대하여 0.1내지 99 중량%를 함유하는 것으로 한다.The present invention provides a cosmetic composition comprising as an active ingredient an extract of U. l. extract prepared according to the present invention or a extract of U. liliaceae. Preferably, the watery extract or watery extract according to the present invention contains 0.1 to 99% by weight based on the total weight of the cosmetic composition.
본 발명의 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 수렴 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포움, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오즈 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 스프레이인 경우에는 클로로플루오로히드로카본, 프로판/부탄 또는 다이메틸 에테르와 같은 추진제를 추가로 함유할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of spraying, chlorofluorohydrocarbons, propane / Or a propellant such as dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용된다.When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 폴리옥시에틸렌 소르비톨 에스테르와 같은 현탁제, 미소결정질 셀룰로오즈 등이 이용될 수 있다.When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as polyoxyethylene sorbitol ester, microcrystalline cellulose or the like may be used as a carrier component.
본 발명의 제형이 계면활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate and the like may be used as a carrier component.
본 발명의 화장료 조성물은 1일에 1회 내지 수회에 걸쳐 얼굴 등에 도포하여 사용할 수 있다.
The cosmetic composition of the present invention can be applied to face or the like once or several times per day.
이하, 본 발명을 하기 실시예 및 실험예를 들어 보다 구체적으로 설명한다. 그러나, 실시예 및 실험예는 본 발명을 예시하기 위한 것일 뿐이므로, 이로써 본 발명의 권리범위가 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, the embodiments and experimental examples are only for illustrating the present invention, and thus the scope of the present invention is not limited thereto.
<< 실시예Example 1> 본 발명 1> invention 율피Julie 추출물 및 율피 발효추출물 제조 Extracts and Production of Fermented Fermented Extracts
본 발명에서 사용한 율피는 대구 소재 (주)푸드웰에서 제공 받아 사용하였으며 율피 발효는 Lactobacillus bifermentans 균주를 이용하였다.
The uri used in the present invention was supplied from Foodwell, Daegu, Korea. The fermented milk was fermented with Lactobacillus bifermentans Were used.
제조예Manufacturing example 1: 본 발명 1: invention 율피Julie 추출물 제조 Extract preparation
율피 열수추출물 (Chestnut inner Shell; CS)은 율피를 시료중량 대비 10배의 물을 가하여 70 ℃에서 3시간씩 3회 반복 열수추출하여 추출물을 수득하였다. 상기 율피 열수추출물은 감압 농축한 다음 동결건조하여 이를 본 발명 율피 열수추출물의 효능검정용 공시재료로 사용하였다.
The extract of Chestnut inner shell (CS) was obtained by adding
제조예Manufacturing example 2: 본 발명 2: invention 율피Julie 발효물Fermentation product 제조 Produce
율피발효에 이용한 L. bifermentans 균주는 MRS(Difco,USA)배지에 배양한 다음 그 배양액을 접종원으로 사용하였다. The L. bifermentans strains used for UFI fermentation were cultured in MRS (Difco, USA) medium and the culture was used as an inoculum.
율피 발효추출물 (Fermented Chestnut inner Shell; FCS)의 제조를 위한 발효배지는 상기 제조예 1에 따라 제조된 율피 열수추출물을 MRS배지 전체에 대하여 0.5중량% 첨가하여 혼합배지를 제조한 다음 상기 L. bifermentans 균주 배양액을 혼합배지 전체에 대하여 1%(v/v)가 되도록 접종하여 24시간 동안 배양하였다. 상기 배양이 끝난 배양액은 6,000 rpm에서 20분 동안 원심분리한 다음 그 상등액을 회수하여 율피발효액을 수득하였다. 상기 율피발효액을 동결건조하여 본 발명 율피 발효추출물을 수득하고 이를 율피 발효추출물의 효능검정용 공시재료로 사용하였다.
The fermentation medium for the preparation of the fermented chestnut inner shell (FCS) was prepared by adding 0.5% by weight of the hydrolyzed water extract prepared according to Preparation Example 1 to the whole MRS medium to prepare a mixed medium, The culture broth was inoculated to the whole of the mixed medium at 1% (v / v) and cultured for 24 hours. The cultured medium Centrifuged at 6,000 rpm for 20 minutes, and the supernatant was recovered to obtain a lupus fermentation broth. The above-described fermentation broth was lyophilized to obtain the fermented fermented extract of the present invention and used as a test material for the evaluation of the efficacy of the fermented fermented extract.
<<
실시예Example
2> 본 발명 2> invention
율피Julie
추출물 및 율피 발효추출물 Extracts and udder fermented extract
의of
효능검정 Efficacy test
실험예Experimental Example 1: 총 폴리페놀 함량 측정 1: Total polyphenol content measurement
총 폴리페놀 함량은 상기 제조예 1 및 제조예 2에 따라 제조된 본 발명 율피 열수추출물(CS)과 율피 발효추출물(FCS)을 Folin - Ciocalteu의 방법에 따라 측정하였다. 본 발명 율피 열수추출물과 율피 발효추출물 각 시료용액을 0.25, 0.5, 0.75, 1 mg/mL 농도로 제조하여 각각 1 mL씩 취하고 1/10으로 희석한 0.2N Folin-Ciocalteu reagent 5 mL와 혼합한 다음 실온에서 5분 동안 반응하였다. 상기 반응이 끝난 반응액에 7.5% sodium carbonate (Na2CO3) 용액 4 mL를 가하여 실온에서 1시간 동안 반응시킨 다음 765 nm에서 흡광도를 측정하였다. 폴리페놀 표준물질 tannic acid를 사용하여 작성한 검량선으로부터 본 발명 율피 열수추출물과 율피 발효추출물에 함유된 폴리페놀 함량을 정량하였으며 모든 실험은 3회 반복하여 실험하였다.Total polyphenol contents of the present invention were measured according to the Folin - Ciocalteu method according to the
총 폴리페놀 함량은 도 1에 나타낸 바와 같이 율피 발효추출물이 율피 열수추출물보다 5 내지 10% 증가했음을 확인하였다
As shown in FIG. 1, the total polyphenol content was increased by 5 to 10% compared to that of the extract of U. P.
실험예Experimental Example 2: 총 플라보노이드 함량 측정 2: Total flavonoid content measurement
총 플라보노이드 함량의 측정은 AlCl₃법에 따라 측정하였다. 본 발명 율피 열수추출물(CS)과 율피 발효추출물(FCS)을 각각 0.25, 0.5, 0.75, 1 mg/mL 농도로 제조한 시료용액 1 mL에 5% sodium nitrite (NaNO2) 용액 150 μL를 혼합하여 실온에서 5분 동안 반응시킨 다음, 10% aluminum chloride (AlCl3) 용액 300 μL를 가하여 실온에서 다시 5분 동안 반응시킨 후 1M NaOH 1 mL와 혼합하여 510 nm에서 흡광도를 측정하였다. 표준물질로 catechin을 사용하여 검량선을 작성하였으며, 모든 실험은 3회 반복하여 실험하였다.Total flavonoid content was measured by AlCl3 method. In the present invention, 1 mL of the sample solution prepared by mixing the hot water extract (CS) and the yeast extract (FCS) at 0.25, 0.5, 0.75 and 1 mg / mL, respectively, was mixed with 150 μL of a 5% sodium nitrite (NaNO 2 ) solution After reacting at room temperature for 5 minutes, 300 μL of 10% aluminum chloride (AlCl 3 ) solution was added, reacted for 5 minutes at room temperature, mixed with 1 mL of 1 M NaOH and absorbance was measured at 510 nm. The calibration curve was prepared using catechin as a reference material. All experiments were repeated three times.
총 플라보노이드 함량은 도 2에 나타낸 바와 같이 본 발명 율피 열수추출물과 율피 발효추출물 사이에 큰 차이는 보이지 않았으나 농도가 높아질수록 플라보노이드 함량이 증가하는 것을 알 수 있었다.
As shown in FIG. 2, there was no significant difference in total flavonoid content between the UFI hot water extract and the UFI fermented extract of the present invention, but it was found that the flavonoid content was increased as the concentration was increased.
실험예Experimental Example 3: 3: 전자공여능Electron donating ability 측정 Measure
본 발명 율피 열수추출물(CS) 및 율피 발효추출물(FCS) 시료용액의 수소전자공여능을 통한 항산화 활성 측정을 위해 DPPH법을 이용하여 측정하였다. DPPH 12 mg을 에탄올 100 mL에 녹인 후 동량의 증류수를 혼합하여 DPPH 용액을 조제하였다. 각 0.25, 0.5, 0.75, 1 mg/mL 농도의 시료용액 0.5 mL에 상기에서 조제한 DPPH 반응용액 5 mL를 넣어 잘 혼합한 후 암소에서 15분 동안 반응시킨 다음 517 nm에서 흡광도를 측정하였다. 수소전자공여능은 각 실험을 3회 반복하여 평균을 낸 다음 대조군에 대한 흡광도의 감소 정도를 하기 계산식 1에 의하여 계산하였다.The DPPH method was used to measure the antioxidative activity of the urinary insecticidal extract (CS) and the uricellium fermented extract (FCS) sample solution by hydrogen electron donating ability. 12 mg of DPPH was dissolved in 100 mL of ethanol, and the same amount of distilled water was mixed to prepare a DPPH solution. bracket 5 mL of the prepared DPPH reaction solution was added to 0.5 mL of the sample solution at 0.25, 0.5, 0.75, and 1 mg / mL, and the mixture was reacted in a dark place for 15 minutes. Then, the absorbance was measured at 517 nm. The hydrogen electron donating ability was calculated by repeating each experiment three times and calculating the degree of decrease in absorbance of the control group according to the following equation.
계산식 1
EDA (%) = (1-A/B) * 100EDA (%) = (1-A / B) * 100
A: 시료 첨가군의 흡광도A: Absorbance of the sample added group
B: 시료 무 첨가군의 흡광도
B: Absorbance of the sample-free group
본 발명 율피 열수추출물 및 율피 발효추출물의 radical 소거 활성을 측정하기 위해 사용된 DPPH radical 소거법은 방향족 화합물 및 방향족 아민류에 의해 환원되어 자색이 탈색에 의해 나타내는 정도를 지표로 하여 항산화능을 측정하는 방법이다.
The DPPH radical scavenging method used for measuring the radical scavenging activity of the UFI hot water extract and the UFI fermented extract of the present invention is a method for measuring the antioxidant ability by reducing the amount of aromatic compounds and aromatic amines .
실험결과 도 3에 나타낸 바와 같이 본 발명 율피 열수추출물과 율피 발효추출물의 free radical 소거활성의 큰 차이는 없으나 율피 열수추출물이 율피 발효추출물에 비해 높은 항산화능을 나타내는 것을 확인할 수 있으며, 율피 열수추출물과 율피 발효추출물 모두 농도 의존적으로 농도가 높아질수록 free radical 소거능이 증가하는 경향을 보였다.
As shown in FIG. 3, there was no significant difference in the free radical scavenging activity between the hot water extract of the present invention and the hot water extract of the present invention. However, the hot water extract of the present invention showed higher antioxidative activity than the hot water extract of the present invention. Free radical scavenging activity tended to increase with increasing concentration of fermented extracts.
실험예Experimental Example 4: 4: SuperoxideSuperoxide anionanion radicalradical 소거능Scatters 측정 Measure
Superoxide anion radical 소거능은 nitroblue tetrazolium (NBT) 환원방법에 따라 실시하였다. 각각 0.25, 0.5, 0.75, 1 mg/mL 농도의 시료용액 0.1 mL에 0.1 M potassium phosphate buffer (pH 7.4) 0.6 mL, 0.4 mM xanthine과 0.24 mM NBT를 녹인 기질액 1 mL를 첨가하고 0.049U/mL xanthine oxidase 1 mL를 가하여 37℃에서 20분 동안 반응시킨 다음 1N HCl 1 mL를 가하여 반응을 종료시키고, 560 nm에서 흡광도 측정하였다. Superoxide anion radical 소거능은 각 실험을 3회 반복하여 평균을 내어 대조군을 이용해 superoxide anion radical 저해율을 하기 계산식 2에 의하여 계산하였다.Superoxide anion radical scavenging activity was measured by nitroblue tetrazolium (NBT) reduction method. each Concentrations of 0.25, 0.5, 0.75, and 1 mg / mL To 0.1 mL of the sample solution, add 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.4), 1 mL of the substrate solution containing 0.4 mM xanthine and 0.24 mM NBT, add 1 mL of 0.049 U / mL xanthine oxidase, and incubate at 37 ° C for 20
계산식 2Equation 2
Superoxide anion radical 저해율(%) = (1-A/B) * 100Superoxide anion radical inhibition rate (%) = (1-A / B) * 100
A: 시료첨가군의 superoxide anion radical 생성량A: Superoxide anion radical production in the sample-added group
B: 무첨가군의 superoxide anion radical 생성량
B: Superoxide anion radical production in the non-additive group
생체 내에서 superoxide radical을 과산화수소로 전환시키는 SOD 유사능의 측정은 NBT법을 이용하였다. The NBT method was used to measure the SOD - like ability to convert superoxide radicals into hydrogen peroxide in vivo.
실험결과 도 4에 나타낸 바와 같이 본 발명 율피 열수추출물(CS)과 율피 발효추출물(FCS) 모두 농도가 높아질수록 superoxide anion radical 소거능이 증가하는 것을 확인할 수 있었으며 본 발명 율피 발효추출물이 율피 열수추출물보다 높은 활성을 나타내는 것을 확인할 수 있다.
As shown in FIG. 4, it was confirmed that the superoxide anion radical scavenging ability of the present invention was increased as the concentration of both the hot water extract (CS) and the fermented hot water extract (FCS) of the present invention was increased. The fermented extract of the present invention showed a higher Activity. ≪ / RTI >
실험예Experimental Example 5: 세포 독성 측정 5: Cytotoxicity measurement
율피 열수추출물과 율피 발효추출물의 세포 독성을 평가하기 위한 HS68 세포는 한국세포주은행(KCLB, Korea Cell Line Bank)에서 분양받았으며, DMEM (Dulbescco's modifiedEagle's media) 와 FBS (Fetal Bovine Serum) 는 WelGENE사(Korea)에서 구입하였고 penicillin-streptomycin은 Gibco사(USA)에서, MTT는 Sigma-Aldrich사(USA)에서 구매하여 사용하였다.HS68 cells were distributed from Korea Cell Line Bank (KCLB) to evaluate the cytotoxicity of UFI hydrolyzate and UFI fermented extract. DMEM (Dulbesco's modified Eagle's media) and FBS (Fetal Bovine Serum) were purchased from WelGENE ), Penicillin-streptomycin was purchased from Gibco (USA), and MTT was purchased from Sigma-Aldrich (USA).
세포의 생존율은 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) 환원 방법을 이용하여 측정하였다. HS68 세포는 10% FBS, 1% penicillin-streptomycin을 포함하는 DMEM 배지를 사용하였다. 세포는 37℃, 5% CO2 조건에서 배양하였다. 세포는 3×104 cell를 24 well plate에 분주하고 37℃, 5% CO2 incubator에서 24시간 동안 배양하였다. 배양한 세포에 율피 열수추출물(CS) 및 율피 발효추출물(FCS)을 0.1 mg/mL 농도로 처리하여 18시간 동안 배양한 후, phosphate buffered saline (PBS) 에 녹인 MTT용액 100 μL를 각 well에 넣고 incubator에서 4시간 동안 배양하였다. 배양 후, 배양액을 버리고 DMSO 1 mL씩 넣어 혼합한 후, 570 nm에서 흡광도를 측정하였다.Cell viability was measured by 3- (4,5-dimethylthiazole-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) reduction method. HS68 cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin. Cells were cultured at 37 ° C and 5% CO 2 . Cells were plated at 3 × 10 4 cells in a 24-well plate and incubated for 24 h at 37 ° C in a 5% CO 2 incubator. After incubation for 18 hours, 100 μL of the MTT solution dissolved in phosphate buffered saline (PBS) was added to each well. incubator for 4 hours. After culturing, the culture medium was discarded, and 1 mL of DMSO was added thereto. The absorbance was measured at 570 nm.
실험결과 도 5에 나타낸 바와 같이 상기 제조예 1 및 제조예 2에 따라 제조된 본 발명 율피 열수추출물 및 율피 발효추출물을 0.1 mg/mL로 처리한 결과 율피 발효추출물이 율피 열수추출물에 비하여 세포 생존율이 증가하였으며 대조군에 비하여 유의적인 세포 독성을 보이지 않았다.
As shown in FIG. 5, when the extracts of the present invention prepared according to Preparation Example 1 and Preparation Example 2 were treated with 0.1 mg / mL of the hot water extract and the hot water extract of the present invention, the cellulase survival rate And did not show significant cytotoxicity compared with the control group.
실험예Experimental Example 6: 6: NitricNitric oxideoxide ( ( NONO ) 생성량 측정) Production amount measurement
율피 열수추출물과 율피 발효추출물의 항염 효과를 평가하기 위한 RAW264.7 대식세포는 한국세포주은행(KCLB, Korea Cell Line Bank)에서 분양받았으며, DMEM (Dulbescco's modifiedEagle's media) 와 FBS (Fetal Bovine Serum) 는 WelGENE사(Korea)에서 구입하였고 penicillin-streptomycin은 Gibco사(USA)에서, Griess Reagent는 Sigma-Aldrich사(USA)에서 구매하여 사용하였다. RAW264.7 macrophages were distributed from Korea Cell Line Bank (KCLB) to evaluate the anti-inflammatory effects of the UFI hydrothermal extract and UFI fermented extract. DME (Dulbesco's modified Eagle's media) and FBS (Fetal Bovine Serum) Griess Reagent was purchased from Sigma-Aldrich (USA) and purchased from Korea. Penicillin-streptomycin was purchased from Gibco (USA) and Griess Reagent was purchased from Sigma-Aldrich (USA).
NO의 농도는 배양액 내의 nitrite 농도를 Griess Reagent System을 이용하여 측정하였다. RAW264.7 대식세포를 5×104 cells/mL의 농도로 24 well plate에 분주하여 24시간 동안 배양한 후에 율피 열수추출물(CS)과 율피 발효추출물(FCS)을 다양한 농도로 처리하여 2시간 동안 배양하였다. 배양 후, LPS 1 μg/mL를 처리하여 18시간 동안 배양하였다. 세포 배양액 100 μL와 Griess 시약 100 μL를 혼합하여 상온에서 15분 동안 반응시킨 후, 540 nm에서 흡광도를 측정하였고, sodium nitrate로 표준 곡선을 작성하여 NO 함량을 산출하였다.NO concentration was measured by using Griess Reagent System. RAW264.7 macrophages were cultured in a 24-well plate at a concentration of 5 × 10 4 cells / mL and cultured for 24 hours. Various concentrations of uricellium pyroxus extract (CS) and uricellium fermented extract (FCS) Lt; / RTI > After culturing, the cells were treated with 1 μg / mL of LPS and cultured for 18 hours. 100 μL of cell culture solution and 100 μL of Griess reagent were mixed and reacted at room temperature for 15 minutes. Absorbance was measured at 540 nm, and a standard curve was prepared with sodium nitrate to calculate the NO content.
실험결과 도 6에 나타낸 바와 같이 RAW264.7 대식세포에 LPS 1 μg/mL를 처리하였을 때 생성된 NO의 함량은 율피 열수추출물 및 율피 발효추출물의 농도가 증가함에 따라 농도의존적으로 감소하는 경향을 보였다. 두 시료의 농도가 0.1 mg/mL에서 율피 발효추출물의 NO 함량은 16.56 μM로 율피 열수추출물에서 생성된 NO의 함량인 17.91 μM에 비해 낮게 나타난 것을 확인하였다.As shown in FIG. 6, the content of NO produced when 1 μg / mL of LPS was treated with RAW 264.7 macrophages tended to decrease in a concentration-dependent manner as the concentration of the hydrolyzate extract and uricellium fermentation extract increased . The NO content of the UFI extract was found to be 16.56 μM at a concentration of 0.1 mg / mL in both samples, which was lower than that of 17.91 μM NO produced in the hot water extract of UFI.
따라서 본 발명 상기 제조예 1 및 제조예 2에 따라 제조된 율피 열수추출물 및 율피 발효추출물은 염증을 개선 또는 치료를 위한 효과를 지니고 있는 것으로 사료된다.
Therefore, it can be concluded that the watery extract and the fermented extract according to the present invention of Preparation Example 1 and Preparation Example 2 have the effect of improving or treating inflammation.
<< 실시예Example 3> 본 발명 3> invention 율피Julie 추출물 및 율피 발효추출물 Extracts and udder fermented extract 의of 항아토피Anti-atopic 활성검정 Active black
아토피를 유발한 동물실험에 실험에 사용한 1-Chloro-2,4-dinitrobenzene (DNCB), Acetone, Olive oil은 Sigma-Aldrich사(USA) 제품을 사용하였으며, Fibrous Tissue Mini Kit,QuantiTect SYBR Green PCR Kit는 QIAgen Korea사(Korea) 제품을, cDNA Sythesis Kit는 Fermentas사(USA) 제품을 사용하여 실험을 수행하였다.For the experiment with atopic dermatitis, 1-Chloro-2,4-dinitrobenzene (DNCB), Acetone and Olive oil were used in Sigma-Aldrich (USA). Fibrous Tissue Mini Kit, QuantiTect SYBR Green PCR Kit Was performed by QIAgen Korea (Korea), and cDNA synthesis kit was used by Fermentas (USA).
실험동물 모델An experimental animal model
실험동물인 6주령의 수컷 NC/Nga mice (21∼26 g)는 (주)중앙실험동물 (Korea) 에서 공급받아 실험 당일까지 사료와 물을 충분히 공급하고 온도 22 ± 2℃, 습도 55 ± 15%, 12시간 주기의 명암을 유지한 실험동물사에서 1주일 동안 적응시킨 후 실험에 사용하였다. Male NC / Nga mice (21-26 g) at 6 weeks of age were supplied from the Central laboratory animal (Korea) and fed with sufficient feed and water until the day of the experiment. The temperature was 22 ± 2 ℃ and the humidity was 55 ± 15 %, And adapted for one week in an experimental animal kept in 12 hour period contrast and used in the experiment.
피부염 유발 및 시료 처리 안정화 기간이 지난 후 NC/Nga mice의 등 부위를 깨끗하게 제모하고 피부의 미세 상처가 치유되도록 24시간 방치하였다. 1% DNCB 용액 (Acetone:Olive oil=3:1) 200 μL를 등 부위에 도포하고, 3일 후 0.4% DNCB 용액 150 μL를 다시 도포하였으며, 4일 후부터는 1주일에 3번씩 0.4% DNCB 용액 150 μL를 3주간 도포하였다. After the induction of dermatitis and stabilization of the sample treatment, NC / Nga mice were gently removed from the dorsal area and left to stand for 24 hours to heal the fine wounds of the skin. 200 μL of a 1% DNCB solution (Acetone: Olive oil = 3: 1) was applied to the back region, and 150 μL of 0.4% DNCB solution was applied again after 3 days. After 4 days, 0.4
실험동물은 총 12마리를 4개의 군으로 나누었으며 정상군(Normal군), 대조군 (AD군)에는 증류수를, 제1실험군(CS)에는 5% 율피 열수추출물을, 제2실험군(FCS)에는 5% 율피 발효추출물을 1주일에 5번씩 3주 동안 도포하였다.
Twelve animals were divided into four groups: normal (normal), control (AD), distilled water, 5% UFI hot water extract in the first experimental group (CS) The 5% UFI fermented extracts were applied for 5 days per week for 3 weeks.
실험예Experimental Example 7: 피부 병변의 평가(육안 관찰) 7: Evaluation of skin lesion (visual observation)
아토피 피부염 유발 후 율피 열수추출물 및 율피 발효추출물을 도포한 실험을 통해 피부조직의 형태학적 변화인 홍반, 건조피부, 짓무름, 부종과 혈종 등을 육안으로 관찰하였다.The morphological changes such as erythema, dry skin, erythema, edema and hematoma of the skin tissue were visually observed through the application of the urophyte hot water extract and the urophyte fermentation extract after the induction of atopic dermatitis.
율피 열수추출물 및 율피 발효추출물이 NC/Nga mice의 피부 병변에 미치는 영향은 DNCB 도포를 통해 아토피 피부염이 유발된 NC/Nga mice에 율피 열수추출물 및 율피 발효추출물을 3주 동안 도포한 후, 도 7과 같이 사진을 찍어 피부 병변의 형태학적 변화를 관찰하였다.The effect of the watery extract of Pucci and the Pucci fermentation extract on the skin lesions of NC / Nga mice was evaluated by applying the Pucci hot water extract and Pucci fermented extract to NC / Nga mice induced by atopic dermatitis through DNCB application for 3 weeks, And the morphological changes of skin lesions were observed.
그 결과, Normal군에서는 피부의 형태학적 변화가 나타나지 않았으나, DNCB를 처리한 AD 군에서는 홍반, 건조피부, 짓무름, 부종과 혈종 등의 아토피성 피부 변화가 확인되었다. 율피 열수추출물을 처리한 CS군에서는 DNCB 도포에 의한 피부의 형태학적 변화가 일부 호전되었지만 큰 차이는 없었으며 율피 발효추출물을 처리한 FCS군에서는 뚜렷한 개선 효과를 보였고 Normal군과 유사한 피부 형태를 나타내었다.
As a result, the morphological changes of the skin were not observed in the normal group, but the atopic skin changes such as erythema, dry skin, erythema, edema and hematoma were observed in the AD group treated with DNCB. The morphological changes of the skin by DNCB application were partially improved in the CS group treated with the hot water extract of URPI, but there was no significant difference, and the FCS group treated with the UF fermented extract showed a marked improvement and showed a skin shape similar to that of the normal group .
실험예Experimental Example 8: 피부 분석 8: Skin analysis
피부 측정 기기인 MPA5 580을 이용하여 아토피 피부염 상태에서 변화하는 피부수분지수, 피부멜라닌지수, 피부홍반지수를 측정 및 비교분석하였다. 시료처치가 끝난 NC/Nga mice의 등 부위 피부를 3번 연속 측정하여 평균값을 얻었으며 측정 장소는 실내온도 21∼23℃, 습도 50∼60%가 유지되는 조건에서 계측하였다.
The skin moisture index, skin melanin index and skin erythema index of atopic dermatitis were measured and compared using MPA5 580. We measured the back skin of NC / Nga mice after 3 consecutive measurements. The measurement was performed at room temperature of 21 ~ 23 ℃ and humidity of 50 ~ 60%.
실험결과 NC/Nga mice의 피부멜라닌지수는 도 6A에 나타낸 바와 같이 Normal군이 44.5 ± 0.17, AD군이 211.33 ± 12.33, CS군이 158.67 ± 23.11, FCS군이 99.56 ± 5.2로 나타나 아토피 피부염 유발로 인해 증가된 멜라닌 함량이 본 발명 율피 발효추출물 도포에 의해 유의적으로 감소되는 것을 확인하였다. As a result, the skin melanin index of NC / Nga mice was 44.5 ± 0.17 in normal group, 211.33 ± 12.33 in AD group, 158.67 ± 23.11 in CS group and 99.56 ± 5.2 in FCS group as shown in FIG. 6A And the melanin content increased due to the addition of the fermented extract of the present invention.
도 6B에 나타낸 피부홍반지수는 Normal군이 156.5 ± 3.83, AD군이 276.33 ± 10.33, CS군이 258.44 ± 26.12, FCS군이 223.22 ± 4.64로 나타나 본 발명 율피 발효물의 도포에 의한 피부 홍반의 감소 경향을 확인할 수 있었다. 이러한 멜라닌 함량과 홍반의 감소 효과는 본 발명 율피열수추출물을 도포하였을 때도 확인할 수 있었지만 상대적으로 본 발명 율피 발효추출물에 의한 감소 효과가 더 두드러지게 나타났다. The skin eruption index shown in FIG. 6B was 156.5 ± 3.83 in the normal group, 276.33 ± 10.33 in the AD group, 258.44 ± 26.12 in the CS group, and 223.22 ± 4.64 in the FCS group, indicating a decrease tendency of skin erythema . The melanin content and the reduction effect of erythema were confirmed by applying the hot water extract of the present invention, but the decrease effect of the hot water extract of the present invention was more remarkable.
피부수분지수도 도 6C에 나타낸 바와 같이 Normal군이 54.6 ± 4.72, AD군이 15.5 ± 2.23, CS군이 11.5 ± 1.04, FCS군이 29.2 ± 2.18로 나타나 DNCB 도포에 의해 감소된 피부의 수분 함량이 본 발명 율피 발효추출물의 도포에 의해 유의적으로 증가되었다. As shown in FIG. 6C, the skin moisture index was 54.6 ± 4.72 in the normal group, 15.5 ± 2.23 in the AD group, 11.5 ± 1.04 in the CS group and 29.2 ± 2.18 in the FCS group, and the skin moisture content decreased by DNCB application Was significantly increased by application of the fermented extract of the present invention.
따라서 상기 실험 결과들은 본 발명 율피 발효추출물이 아토피 피부질환에서 나타나는 피부 병변의 변화를 호전시킬 수 있음을 확인하였다.
Therefore, the above experimental results confirm that the present invention can improve the skin lesion change in atopic skin diseases.
실험예Experimental Example 9: 염증유발 유전자 발현 분석 9: Analysis of inflammatory gene expression
염증 사이토카인의 유전자 발현 측정 피부조직의 RNA는 Fibrous Tissue Mini Kit를 이용하여 추출하였다. 추출된 RNA의 농도는 NanoDrop ND-1000을 이용하여 측정하였으며 cDNA Synthesis Kit를 이용하여 5 μg의 RNA를 cDNA로 합성한 후, real-time PCR에 사용하였다. Real-time PCR은 Corbett research RG-6000을 이용하여 수행하였다. 염증 사이토카인 유전자 발현은 QuantiTect SYBR Green PCR Kit를 사용하였고, endogenous control은 GAPDH를 사용하였으며, primer의 최종 농도가 10 pM이 되게 반응시켰다. Real-time PCR에 사용된 염증 사이토카인 primer의 염기배열은 하기 [표 1]과 같다.Measurement of gene expression of inflammatory cytokines RNA of skin tissue was extracted using Fibrous Tissue Mini Kit. The concentration of extracted RNA was measured using NanoDrop ND-1000. 5 μg of RNA was synthesized by cDNA synthesis kit and then used for real-time PCR. Real-time PCR was performed using Corbett research RG-6000. The expression of the inflammatory cytokine gene was quantitated using the SYBR Green QuantiTect PCR Kit, the endogenous control was performed using GAPDH, and the final concentration of the primer was 10 pM. The nucleotide sequences of the inflammatory cytokine primers used in the real-time PCR are shown in Table 1 below.
실험결과 앞의 실험을 통해 확인된 본 발명 율피 발효추출물의 항아토피 효능이 피부에 존재하는 염증 사이토카인 유전자의 변화와 관련 있는지 확인하기 위해 IL-1β와 TNF-α에 대한 quantitative real-time PCR을 수행하였다. Experimental Results The quantitative real-time PCR of IL-1β and TNF-α was performed to confirm whether the anti-atopic effect of the present invention was related to the change of the inflammatory cytokine gene present in the skin. Respectively.
피부에서 IL-1β mRNA 유전자 발현은 도 7A에 나타낸 바와 같이 Normal군의 RQ 값이 1일 때, AD군은 1.34로 아토피 유발로 인해 IL-1β mRNA 발현이 증가되었고 CS군은 0.95, FCS군은 0.82로 나타나 AD군에 비하여 감소되는 경향을 나타내었다. As shown in FIG. 7A, when the RQ value of the normal group was 1, expression of IL-1 beta mRNA was 1.34 in the AD group, IL-1 beta mRNA expression was increased due to atopy, 0.95 in the CS group, 0.82, indicating a tendency to decrease compared to the AD group.
TNF-αmRNA 유전자 발현은 도 7B에 나타낸 바와 같이 Normal군의 RQ값이 1일 때, AD군 1.37, CS군 1.1, FCS군 1.06으로 나타나 AD군에서 증가된 TNF-α mRNA의 발현이 본 발명 율피열수추출물 및 율피 발효물 도포에 의해 감소되는 경향을 나타내었다. As shown in FIG. 7B, when the RQ value of the normal group was 1, expression of TNF-α mRNA gene was 1.37 in the AD group, 1.1 in the CS group, and 1.06 in the FCS group, And decreased by application of hot - water extract and UFI fermented product.
따라서 본 발명 율피열수추출물 및 율피 발효추출물이 IL-1β, TNF-α와 같은 염증 사이토카인의 발현을 감소시킴으로써 아토피 피부염에 치료 효과를 나타냄을 확인할 수 있었다.
Thus, it was confirmed that the present invention of the present invention is effective in reducing atopic dermatitis by reducing the expression of inflammatory cytokines such as IL-1β and TNF-α.
이상 설명한 바와 같이 본 발명은 율피 열수추출물 또는 율피 발효추출물을 함유하는 알레르기성 피부질환 개선, 예방 및 치료용 조성물을 제공하는 뛰어난 효과가 있으므로 천연물의약소재산업상 매우 유용한 발명이다.
As described above, the present invention is an extremely useful invention in the natural medicinal materials industry because it has an excellent effect of providing a composition for the improvement, prevention and treatment of allergic skin disease containing a hot water extract of U.
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KR20160072382A (en) * | 2014-12-12 | 2016-06-23 | 주식회사 명진에프앤비 | Method for producing CHESTNUT INNER SKIN fermented beverage and CHESTNUT INNER SKIN fermented beverage produced by the same |
KR101881369B1 (en) | 2017-02-21 | 2018-07-24 | 충남대학교산학협력단 | Anti-oxidant Aged Chestnut Inner Shell and Anti-oxidant Composition Comprising the Aged Chentnut Inner Shell or Its Extract |
KR20180102890A (en) | 2017-03-08 | 2018-09-18 | 충남대학교산학협력단 | Anti-Oxidant Yanggeng Containing Aged Chestnut Inner shell |
KR20210157328A (en) * | 2020-06-19 | 2021-12-28 | 고려대학교 산학협력단 | Cosmetic compositions comprising fermented chestnut inner shell extracts or its fractions as effective components |
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KR20180102890A (en) | 2017-03-08 | 2018-09-18 | 충남대학교산학협력단 | Anti-Oxidant Yanggeng Containing Aged Chestnut Inner shell |
KR20210157328A (en) * | 2020-06-19 | 2021-12-28 | 고려대학교 산학협력단 | Cosmetic compositions comprising fermented chestnut inner shell extracts or its fractions as effective components |
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