KR20130077317A - Functional food composition for improving skin condition and preparation method thereof - Google Patents
Functional food composition for improving skin condition and preparation method thereof Download PDFInfo
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- KR20130077317A KR20130077317A KR1020110145959A KR20110145959A KR20130077317A KR 20130077317 A KR20130077317 A KR 20130077317A KR 1020110145959 A KR1020110145959 A KR 1020110145959A KR 20110145959 A KR20110145959 A KR 20110145959A KR 20130077317 A KR20130077317 A KR 20130077317A
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- extract
- food composition
- functional food
- skin
- health functional
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/40—Shaping or working of foodstuffs characterised by the products free-flowing powder or instant powder, i.e. powder which is reconstituted rapidly when liquid is added
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2250/00—Food ingredients
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- Engineering & Computer Science (AREA)
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Abstract
Description
본 발명은 피부 질환 개선용 건강기능식품 조성물 및 그 제조방법에 관한 것으로, 보다 구체적으로는 맥문동, 인삼, 오미자 및 어성초의 추출물을 포함하는 피부 질환 개선용 건강기능식품 조성물 및 그 제조방법에 관한 것이다.
The present invention relates to a health functional food composition for improving skin diseases, and a method of manufacturing the same, and more particularly, to a health functional food composition for improving skin diseases, including extracts of Macmundong, ginseng, Schisandra chinensis and Estuary. .
인체의 피부는 물리적, 화학적으로 외계로부터 신체를 보호하는 동시에 전신의 대사에 필요한 생화학적 기능을 영위하는 생명유지에 필수 불가결한 기관이다. 비교적 흔한 피부질환에는 아토피 피부염, 접촉피부염, 지루성 피부염, 두드러기, 무좀, 완선, 건선, 단순포진/대상포진, 피부건조증, 주부습진, 여드름 등이 있다.The skin of the human body is an indispensable organ for the maintenance of life, which physically and chemically protects the body from extraterrestrial and performs the biochemical functions necessary for metabolism of the whole body. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, psoriasis, psoriasis, herpes simplex / shingles, dry skin, housewives and acne.
이러한 피부질환의 외인으로는 외계로부터 직접 피부에 작용하는 마찰, 압박, 타박 등의 기계적 자극을 비롯하여, 동물성, 식물성 및 광물성의 독물, 그 밖에 약품, 방사선, 온열, 한랭, 또는 세균, 사상균, 원충, 바이러스 등의 미생물이나 피부기생동물 등을 들 수 있으며, 내인으로는 위장장애, 간장병, 신장병, 물질대사장애, 혈액질환, 내장종양, 내분비장애 등을 들 수 있다. 유전성 피부질환 및 모반, 모반증 등은 그 원인이 유전자에 의한 것이므로 내인성 피부병에 포함된다.The external causes of such skin diseases include mechanical stimuli such as friction, compression, and bruise, which act directly on the skin from the outside world, as well as animal, vegetable and mineral poisons, other medicines, radiation, heat, cold, or bacteria, filamentous fungi, protozoa And microorganisms such as viruses and skin parasitic animals, and the like, and endogenous diseases include gastrointestinal disorders, liver disease, kidney disease, metabolic disorders, blood diseases, visceral tumors, and endocrine disorders. Hereditary skin diseases, birthmarks, birthmarks and the like are caused by genes and are included in endogenous skin diseases.
피부질환 중, 특히 아토피 피부염은 아토피 알레르기를 가진 사람에게서 나타나는 대표적인 피부질환으로서, 흔히 태열이라 불리는 만성피부질환의 하나이며 심한 소양감을 동반하는 재발성 만성 피부염이다. 최근 수십년 동안 환경오염 등에 의한 공기 매개성 알레르기 유발물질(allergen)의 증가와 건조성 피부를 형성시키는 조건의 증가 등으로 인하여 아토피 피부염에 대한 유병율이 계속 상승하고 있으며, 전 세계 인구의 10 내지 20%가 이 피부염으로 고통받고 있는 것으로 알려져 있다. 아토피 피부염은 주로 유아와 소아기에 발병하여 성인기까지 지속되는 만성 재발성 피부염으로, 극심한 소양감과 건조 피부, 홍반, 부종, 삼출 등을 동반하는 피부질환이다. 국내 아토피 피부염의 유병율에 대한 한 연구에 따르면, 소아의 경우 아토피 피부염으로 진단받은 유병율을 28.22%로 보고하였고, 다른 연구에서는 성인 아토피 피부염의 자각적 유병율을 약 3.09%로 보고한 바 있다. 주증상은 가려움증 및 피부 건조증이고, 면역학적 특성을 보여 다른 알레르기 질환인 두드러기, 천식, 알레르기성 비염, 알레르기성 결막염 등을 동반하는 경우도 있다. Among skin diseases, in particular, atopic dermatitis is a representative skin disease in people with atopic dermatitis, and is one of chronic skin diseases commonly referred to as febrile fever and recurrent chronic dermatitis with severe pruritus. In recent decades, the prevalence of atopic dermatitis continues to increase due to the increase in airborne allergens due to environmental pollution and the increase in conditions for forming dry skin. It is known that% suffer from this dermatitis. Atopic dermatitis is a chronic relapsing dermatitis that occurs mainly in infants and young children and lasts to adulthood. It is a skin disease with severe pruritus and dry skin, erythema, edema, and exudation. According to a study on the prevalence of atopic dermatitis in Korea, 28.22% reported the prevalence of atopic dermatitis in children, and another study reported the subjective prevalence of adult atopic dermatitis as about 3.09%. Its main symptoms are itching and dry skin, and its immunological properties may lead to other allergic diseases such as urticaria, asthma, allergic rhinitis, and allergic conjunctivitis.
또한 세포의 노화는 생리적 자연현상으로서, 피부를 구성하고 있는 기본 단위인 피부 세포의 분열이 정상적으로 이루어지지 않고 재생되는 기간이 점점 길어지게 되며, 그 결과 피부는 탄력을 잃고, 노화 각질이 쌓이고 건조해지면서 주름이 생기기 시작하면서 진행된다. 현재, 노화로 인한 피부를 회복시키기 위해 수술이나 연고 및 비타민 C 요법 등을 사용하고 있지만 우리의 오랜 숙원인 팽팽하고 탄력 있는 피부를 유지하기는 역부족이다.In addition, aging of cells is a physiological natural phenomenon, and the period of regeneration of skin cells, which are the basic units constituting the skin, is not normal, and the regeneration period becomes longer. It progresses as it begins to wrinkle. Currently, surgery, ointment, and vitamin C therapy are used to restore the skin caused by aging, but it is not enough to maintain the long and tender skin.
이러한 아토피성 피부질환을 개선시키거나 치료하기 위한 방법으로는 피부 건조 증상을 개선하기 위한 방법들, IgE의 생성을 억제하는 면역 조절제를 사용하는 방법, 항균 및/또는 항염증 성분들을 이용하는 방법, 가려움증을 개선하는 방법 등이 있고 이 중에서 두 가지 이상을 복합하는 방법도 알려져 있다. 또한 아토피 피부염, 염증성 피부염 등을 포함하는 피부질환의 치료에는 보통 스테로이드제가 널리 사용되고 있으나, 스테로이드제는 면역기능 저하, 피부의 박화나 위축, 홍조 등의 다양한 부작용을 발현할 수 있고, 장기 복용시에는 스테로이드성 여드름, 스테로이드성 피부(피부위축, 모세혈관 확장, 자반), 스테로이드성 주사, 입주위 피부염, 다모, 색소탈실, 선조, 수포성 피부염, 피부출혈 등이 나타날 수 있는 등 심각한 부작용이 보고되고 있다. Methods for improving or treating such atopic skin diseases include methods for improving skin dryness symptoms, using immunomodulators that inhibit the production of IgE, using antibacterial and / or anti-inflammatory components, itching There is a method to improve the and the like and a method of combining two or more of these is also known. In addition, steroids are widely used in the treatment of skin diseases including atopic dermatitis, inflammatory dermatitis, etc. However, steroids can cause various side effects such as decreased immune function, thinning or atrophy of the skin, and flushing. Serious side effects have been reported, such as steroid acne, steroid skin (skin atrophy, capillary enlargement, purpura), steroid injections, perioral dermatitis, hairyness, pigment loss, ancestors, bullous dermatitis, skin bleeding, etc. have.
몇 년 전부터 천연물을 이용한 피부 노화 방지 및 피부 개선제 등이 개발되고 있으나, 그 중에는 남성호르몬인 안드로겐(androgen) 억제작용 및 그에 따른 피지분비증가 억제 작용에 대한 피부 개선제로서의 기능 등을 가진 경우도 있고, 피부에 대한 치료제로서의 실질적인 효과를 제공하지는 못하고 있는 실정이다. 최근에는 메디칼 스킨 케어(medical skin care)라 해서, 의학적 치료와 스킨 케어가 접목된 피부 치료방법이 우리나라뿐만 아니라 미국, 유럽, 일본 등지에서도 피부 치료를 위해 개발되고 있으나, 이 방법은 알레르기 피부질환, 여드름, 기미, 주근깨 등을 개선시킬 뿐 여전히 많은 피부질환은 극복되지 않은 채 삶의 질을 저하시키는 중요한 원인이 되고 있다.Several years ago, anti-aging and skin-improving agents using natural products have been developed, but some of them have a function of inhibiting androgen (androgen), which is a male hormone, and a skin-improving effect on sebum secretion. It does not provide a practical effect as a treatment for the skin. Recently, medical skin care (medical skin care), a combination of medical treatment and skin care skin treatment method has been developed for the skin treatment not only in Korea, but also in the United States, Europe, Japan, etc. In addition to improving acne, blemishes and freckles, many skin diseases are still a major cause of poor quality of life without being overcome.
이와 관련하여 피부 개선, 보습, 주름개선 등의 효과를 갖는 것으로 알려진 생약들의 유효성분을 추출하여 피부 개선제로 개발된 예들이 알려져 있다. 예를 들어, 한국공개특허 제10-2010-0101269호, 한국등록특허 제10-0962587호에는 매우 다양한 식물 소재를 사용한 피부 개선용 조성물을 제공하고 있으나, 양자 모두 발효 방법에 특징이 있는 기술로서, 특정 식물의 단독 또는 혼합 추출물의 우수한 피부 개선 효과를 본 것이 아니다. 또한 한국등록특허 제10-0991399호, 한국등록특허 제10-0999870호, 한국등록특허 제10-0987563호 등에는 매우 다양한 식물 추출물을 포함하는 아토피 피부염 개선용 조성물이 기재되어 있다.In this regard, there are known examples of extracts of active ingredients of herbal medicines known to have effects such as skin improvement, moisturization, wrinkle improvement, and the like, and have been developed as skin improvers. For example, Korean Patent Publication No. 10-2010-0101269 and Korean Patent Registration No. 10-0962587 provide a skin improvement composition using a wide variety of plant materials, both of which are characterized by the fermentation method, It is not seen that the skin improvement effect of a single or mixed extract of a particular plant is excellent. In addition, Korean Patent Registration No. 10-0991399, Korean Patent Registration No. 10-0999870, Korean Patent Registration No. 10-0987563, etc., describes a composition for improving atopic dermatitis including a wide variety of plant extracts.
그러나 이러한 기술은 대부분 가려움증이나 염증 등을 경감시켜 줄 뿐이어서 그 치료효과가 미미한 문제점이 있어 실제로는 그 적용에 한계가 있는 실정이다. 앞서 기술한 아토피 피부염을 치유하거나 개선하는 방법들은 가려움증 저감 또는 항염증 등 한두 가지의 증상에 대한 완화나 개선을 목적으로 하며 복합 감염 등 심각한 부작용이 올 수도 있으며 인공합성 물질의 사용으로 증세가 악화되는 경우도 발생하고 있다. 또한, 여러 가지 한약재를 포함하는 것들은 주로 경구용 약제 또는 건강보조식품으로 아토피 피부염 완화에 이로울 수 있으나 다른 부작용이 발생할 수 있다. 즉, 여러 가지 천연 한약 재료를 이용한 경우에는 부작용의 개선 방법이 상호 다를 수도 있고 호전되는 시기가 각기 다르며 재발 방지 효과도 서로 다를 수 있다. 따라서 우수한 효과를 가지면서도 피부 자극이 없고 안전하게 조성한 한방 조성물이 요망된다.
However, most of these techniques only relieve itching and inflammation, so the therapeutic effect is insignificant, and the practical application of the present invention is limited. The aforementioned methods for treating or improving atopic dermatitis aim to alleviate or improve one or two symptoms, such as itching or anti-inflammatory, and may have serious side effects such as complex infections. It is also happening. In addition, those containing various herbal medicines are mainly oral medications or health supplements may be beneficial to relieve atopic dermatitis, but other side effects may occur. In other words, when using a variety of natural herbal ingredients, the way to improve the side effects may be different from each other, the timing of improvement is different, and the relapse prevention effect may also be different. Therefore, there is a need for a herbal composition that has excellent effects and has no skin irritation and is safely prepared.
본 발명을 통하여 건조 피부의 보습력 향상, 아토피 증상 개선, 알레르기 증상 개선, 주름 개선, 피부톤 향상, 미백, 가려움증 개선, 피부 장벽기능 개선, 피부 면역기능 향상 및 염증 증상 개선이 우수하면서도 안전성이 뛰어나고 부작용이 적은 피부 질환 개선용 건강기능식품 조성물 및 그 제조방법을 제공한다.
Improved moisturizing ability of dry skin, atopic symptoms improvement, allergy symptoms improvement, wrinkle improvement, skin tone improvement, whitening, itching improvement, skin barrier function improvement, skin immune function improvement and inflammatory symptoms improvement but excellent safety and side effects The present invention provides a health functional food composition for reducing skin diseases and a method of manufacturing the same.
본 발명의 일 태양에 따르면, 맥문동, 인삼, 오미자 및 어성초의 추출물을 포함하는 피부 질환 개선용 건강기능식품 조성물을 제공한다. According to an aspect of the present invention, there is provided a health functional food composition for improving skin diseases, including extracts of mammundong, ginseng, Schisandra chinensis and Eochocho.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 추출물은 조추출액, 조추출액의 건조물, 조추출액의 농축물. 조추출액의 분획물, 분획물의 건조물 및 분획물의 농축물 중에서 선택된 하나 이상인 것을 특징으로 한다.According to one aspect of the invention, the extract of the health functional food composition for improving skin diseases is crude extract, dried extract of crude extract, concentrate of crude extract. It is characterized in that at least one selected from fractions of crude extract, dried fractions and concentrates of fractions.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 맥문동, 인삼, 오미자 및 어성초의 추출물은 맥문동 : 인삼 : 오미자 : 어성초의 중량비율이 1 내지 8 : 1 내지 4 : 1 내지 4 : 1 내지 8 인 것을 특징으로 한다.According to one aspect of the present invention, the extract of McMung-Dong, Ginseng, Schisandra chinensis and Eochochocho of the health functional food composition for improving skin diseases is the weight ratio of Macmundong: Ginseng: Schisandra chinensis: Echochocho to 1-8: 1-4: 1-4. : It is characterized by 1-8.
본 발명의 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 맥문동, 인삼, 오미자 및 어성초의 추출물은 맥문동 : 인삼 : 오미자 : 어성초의 중량비율이 1 : 1 : 1 : 2 인 것을 특징으로 한다.According to an aspect of the present invention, the extracts of McMun-Dong, Ginseng, Schisandra chinensis and Eochochocho of the health functional food composition for improving skin diseases are characterized in that the weight ratio of McMoon-Dong: Ginseng: Schisandra chinensis: Echochocho is 1: 1: 1: 1. .
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 추출물은 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 한다.According to an aspect of the present invention, the extract of the health functional food composition for improving skin diseases is characterized in that the extract is available as a solvent selected from water, lower alcohols having 1 to 4 carbon atoms or a mixed solvent thereof.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물은 상기 추출물에 담체, 희석제, 부형제 및 첨가제 중 하나 이상을 더 포함하여 정제, 환제, 산제, 과립제, 분말제, 캡슐제 및 액제 제형으로 이루어진 군에서 선택된 하나로 제형된 것을 특징으로 한다.According to an aspect of the present invention, the health functional food composition for improving skin diseases further comprises at least one of a carrier, a diluent, an excipient and an additive in the extract, tablets, pills, powders, granules, powders, capsules and liquids. It is characterized by being formulated as one selected from the group consisting of formulations.
본 발명의 일 태양에 따르면, 맥문동, 인삼, 오미자 및 어성초에 용매를 가하여 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함하는 피부 질환 개선용 건강기능식품 조성물의 제조방법을 제공한다.According to an aspect of the present invention, the step of extracting by adding a solvent to ganmundong, ginseng, Schisandra chinensis and Eochocho to obtain an extract; And it provides a method for producing a health functional food composition for improving skin diseases comprising the step of concentrating the extract.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 제조방법의 추출물을 수득하는 단계는 맥문동, 인삼, 오미자 및 어성초의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출되는 것을 특징으로 한다.According to one aspect of the invention, the step of obtaining an extract of the method for producing a health functional food composition for improving skin diseases is extracted by adding a solvent of 5 to 20 times the weight of the total weight of ganmundong, ginseng, Schisandra chinensis and Echochocho It features.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 제조방법의 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 수행되는 것을 특징으로 한다.According to one aspect of the invention, the step of obtaining the extract of the method for producing a health functional food composition for improving skin diseases is characterized in that it is performed for 1 hour to 10 hours at 80 ℃ to 120 ℃.
본 발명의 일 태양에 따르면, 상기 피부 질환 개선용 건강기능식품 조성물의 제조방법의 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 한다.
According to one aspect of the invention, the step of concentrating the manufacturing method of the health functional food composition for improving skin diseases comprises the steps of filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; It characterized in that it comprises the step of pulverizing the dried concentrate concentrate.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물은 건조 피부의 보습력 향상, 아토피 피부염 증상 개선, 알레르기 증상 개선, 주름 개선, 피부톤 향상, 미백, 가려움증 개선, 피부 장벽기능 향상, 피부 면역 기능 향상 및 염증 증상 개선이 우수하면서도 안전성이 뛰어나고 부작용이 적은 효과를 갖는다.The health functional food composition for improving skin diseases according to the present invention improves the moisturizing ability of dry skin, improves atopic dermatitis symptoms, improves allergic symptoms, improves wrinkles, improves skin tone, improves whitening, itching, improves skin barrier function, improves skin immune function and inflammation. Excellent symptom improvement but safety and fewer side effects.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 제조방법은 건조 피부의 보습력 향상, 아토피 증상 개선, 알레르기 증상 개선, 주름 개선, 피부톤 향상, 미백, 가려움증 개선, 피부 장벽기능 향상, 피부 면역 기능 향상 및 염증 증상 개선이 우수하면서도 안전성이 뛰어나고 부작용이 적은 피부 질환 개선용 건강기능식품 조성물을 효과적으로 제공한다.
The method for producing a health functional food composition for improving skin diseases according to the present invention improves the moisturizing ability of dry skin, improves atopic symptoms, improves allergic symptoms, improves wrinkles, improves skin tone, improves whitening, itching, improves skin barrier function, and improves skin immune function. And it provides an excellent functional food composition for improving skin diseases with excellent safety and excellent safety and less side effects.
도 1은 실시예 1 및 비교예를 농도별로 처리하여 세포 배양한 결과이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 2는 실시예 1 및 비교예를 농도별로 처리한 경우, 타이로시네이즈 저해 활성을 나타낸 그래프이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 3은 실시예 1 및 비교예를 농도별로 처리한 경우, 필라그린 및 SPT의 발현 정도를 나타낸 western blotting사진이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 4는 실시예 1 및 비교예를 농도별로 처리한 경우, 필라그린 및 SPT의 발현 정도를 상대적으로 도시한 그래프이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 5는 실시예 1 및 비교예를 농도별로 처리한 경우, COX-2 및 AP-1의 발현 정도를 나타낸 western blotting사진이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 6은 실시예 1 및 비교예를 농도별로 처리한 경우, COX-2 및 AP-1의 발현 정도를 상대적으로 도시한 그래프이다. (a)는 비교예, (b)는 실시예 1을 처리한 결과이다.
도 7은 아토피 피부염을 유발한 동물 실험에서, 실험 초기의 각 군별 동물의 피부 상태를 나타낸 사진이다. A는 control(미투여), B는 비교군(비교예 투여), C는 실험군(실시예1 투여)이다.
도 8은 아토피 피부염을 유발한 동물 실험에서, 투여 1주 차의 각 군별 동물의 피부 상태를 나타낸 사진이다. A는 control(미투여), B는 비교군(비교예 투여), C는 실험군(실시예1 투여)이다.
도 9는 아토피 피부염을 유발한 동물 실험에서, 투여 2주 차의 각 군별 동물의 피부 상태를 나타낸 사진이다. A는 control(미투여), B는 비교군(비교예 투여), C는 실험군(실시예1 투여)이다.
도 10은 아토피 피부염을 유발한 동물 실험에서, 투여 3주 차의 각 군별 동물의 피부 상태를 나타낸 사진이다. A는 control(미투여), B는 비교군(비교예 투여), C는 실험군(실시예1 투여)이다.
도 11은 아토피 피부염을 유발한 동물 실험에서, 투여 4주 차의 각 군별 동물의 피부 상태를 나타낸 사진이다. A는 control(미투여), B는 비교군(비교예 투여), C는 실험군(실시예1 투여)이다.
도 12는 아토피 피부염을 유발한 동물 실험에서, 투여 기간에 따른 피부 염증 정도를 점수화하여 나타낸 그래프이다.
도 13은 아토피 피부염을 유발한 동물 실험에서, 투여 기간에 따른 면역글로불린 E(IgE)의 양을 나타낸 그래프이다.
도 14는 아토피 피부염을 유발한 동물 실험에서, 인터루킨(IL)-4의 양을 나타낸 그래프이다.
도 15는 아토피 피부염을 유발한 동물 실험에서, 인터루킨(IL)-5의 양을 나타낸 그래프이다.
도 16은 아토피 피부염을 유발한 동물 실험에서, 인터루킨(IL)-6의 양을 나타낸 그래프이다.
도 17은 아토피 피부염을 유발한 동물 실험에서, 면역글로불린M(IgM)의 양을 나타낸 그래프이다.
도 18은 아토피 피부염을 유발한 동물 실험에서, 면역글로불린G1(IgG1)6의 양을 나타낸 그래프이다.
도 19는 아토피 피부염을 유발한 동물 실험에서, 인터페론감마(IFN-γ)의 양을 나타낸 그래프이다.1 is a result of cell culture by treating Example 1 and Comparative Example by concentration. (a) is a comparative example, (b) is the result of having processed Example 1.
2 is a graph showing the tyrosinase inhibitory activity when Example 1 and Comparative Example were treated by concentration. (a) is a comparative example, (b) is the result of having processed Example 1.
3 is a western blotting photograph showing the expression levels of filaggrin and SPT when treated according to the concentration of Example 1 and Comparative Examples. (a) is a comparative example, (b) is the result of having processed Example 1.
FIG. 4 is a graph showing relatively the expression levels of filaggrin and SPT when Example 1 and Comparative Example are treated by concentration. (a) is a comparative example, (b) is the result of having processed Example 1.
5 is a western blotting photograph showing the expression level of COX-2 and AP-1 when Example 1 and Comparative Example were treated by concentration. (a) is a comparative example, (b) is the result of having processed Example 1.
FIG. 6 is a graph showing relatively the expression level of COX-2 and AP-1 when Example 1 and Comparative Example were treated by concentration. (a) is a comparative example, (b) is the result of having processed Example 1.
Figure 7 is a photograph showing the skin condition of the animals in each group at the beginning of the experiment in the animal experiment causing atopic dermatitis. A is control (not administered), B is comparative group (comparative administration), C is experimental group (Example 1 administration).
Figure 8 is a photograph showing the skin condition of the animals in each group of the first week of administration in the animal experiments causing atopic dermatitis. A is control (not administered), B is comparative group (comparative administration), C is experimental group (Example 1 administration).
Figure 9 is a photograph showing the skin condition of animals in each group at the 2nd week of administration in the animal experiments inducing atopic dermatitis. A is control (not administered), B is comparative group (comparative administration), C is experimental group (Example 1 administration).
10 is a photograph showing the skin condition of animals in each group at the 3rd week of administration in an animal experiment inducing atopic dermatitis. A is control (not administered), B is comparative group (comparative administration), C is experimental group (Example 1 administration).
11 is a photograph showing the skin condition of animals in each group at the 4th week of administration in an animal experiment inducing atopic dermatitis. A is control (not administered), B is comparative group (comparative administration), C is experimental group (Example 1 administration).
12 is a graph showing the score of the degree of skin inflammation according to the administration period in the animal experiments inducing atopic dermatitis.
FIG. 13 is a graph showing the amount of immunoglobulin E (IgE) according to the administration period in animal experiments causing atopic dermatitis.
14 is a graph showing the amount of interleukin (IL) -4 in animal experiments that induce atopic dermatitis.
FIG. 15 is a graph showing the amount of interleukin (IL) -5 in animal experiments causing atopic dermatitis.
Figure 16 is a graph showing the amount of interleukin (IL) -6 in animal experiments causing atopic dermatitis.
Figure 17 is a graph showing the amount of immunoglobulin M (IgM) in animal experiments causing atopic dermatitis.
Figure 18 is a graph showing the amount of immunoglobulin G1 (IgG1) 6 in animal experiments causing atopic dermatitis.
Figure 19 is a graph showing the amount of interferon gamma (IFN-γ) in animal experiments causing atopic dermatitis.
상기 상술한 목적을 달성하기 위한 본 발명은 맥문동, 인삼, 오미자 및 어성초의 추출물을 포함하는 피부 질환 개선용 건강기능식품 조성물과 그 제조방법을 제공한다. 이하 첨부된 도면을 이용하여 본 발명의 구성을 더욱 상세하게 살펴보도록 한다.The present invention for achieving the above-described object provides a health functional food composition for improving skin diseases, including the extract of the mammundong, ginseng, Schisandra chinensis and Eochochocho and a method for producing the same. Hereinafter, the configuration of the present invention will be described in detail with reference to the accompanying drawings.
본 발명에 따르면, 맥문동, 인삼, 오미자 및 어성초의 추출물을 포함하는 피부 질환 개선용 건강기능식품 조성물을 제공한다.According to the present invention, there is provided a health functional food composition for improving skin diseases, including extracts of mammundong, ginseng, Schisandra chinensis and Echochocho.
맥문동(Liriope platyphyllla)은 소엽맥문동 및 동속근연 식물의 괴근으로, 외떡잎식물 백합목 백합과의 상록으로 그늘진 곳에서 자란다. 맛은 감미고하고 성질은 차다. 폐, 위, 심경에 들어간다. 주로 소염, 강장, 진해, 거담제 및 강심제로 사용한다. 맥문동의 괴근에는 여러 종류의 스테로이드계 사포닌(saponin)이 함유되어 있고, 그 아글리콘(aglycone)은 ruscogenin이다. 또 베타시토스테롤(β-sitosterol), 스티그마스테롤(stigmasterol), 베타시토스테롤베타디글루코사이드(β-sitosterol-β-d-glucoside)를 함유하고 있다. 맥문동의 과실에는 캠프페롤-3-글루코갈락토사이드(kaempferol-3-glucogalactoside)라는 오피오사이드(ophioside)를 함유하고 있다. 맥문동은 혈당을 낮추며, 백색포도구균, 대장균에 대하여도 항균작용이 있는 것으로 알려져 있다.Limoope platyphyllla is a lump of lobular vegetation and related plants and grows in the shade of evergreens of the monocotyledonous lily. The taste is sweet and the nature is cold. Enter the lungs, stomach and heart. It is mainly used as anti-inflammatory, tonic, cough, expectorant and cardiac. The muscle mass of the pulmonary sinus contains various types of steroidal saponins, and its aglycone is ruscogenin. It also contains beta sitosterol (β-sitosterol), stigmasterol (stigmasterol), and beta sitosterol beta diglucoside (β-sitosterol-β-d-glucoside). The fruit of the pulses contains an opioside called kaempferol-3-glucogalactoside. Macmundong lowers blood sugar and is known to have antimicrobial activity against white Pococcus and E. coli.
인삼(Panax ginseng)은 오가피과 인삼속 식물인 인삼의 뿌리를 건조한 것으로 신농본초경에서는 기를 크게 보하고 비장과 페장의 기운을 북돋우며, 갈증을 멎게하고, 정신을 평안하게 하는 효능이 있다고 알려져 있다. 또한, 현대 약리학적으로는 중추신경계 조절, 면역증강, 강심, 혈당강하 등의 효능이 있다고 한다.Ginseng (Panax ginseng) is the root of ginseng, a genus of ginseng and ginseng plant, and is known to be effective in reinforcing the ginseng roots, encouraging the spleen and the intestines, quenching thirst, and calming the mind. In addition, modern pharmacological effects such as central nervous system control, immune enhancement, heart rate, blood sugar lowering effect is said.
오미자는 오미자(Schizandra chinensis Baillon)나무의 열매로 지름 약 1cm의 공 모양이며, 짙은 붉은 빛깔이다. 특징은 단맛·신맛·쓴맛·짠맛·매운맛의 5가지 맛이 나며, 그 중에서도 신맛이 가장 강하다. 오미자는 시잔드린, 고미신, 시트럴, 사과산, 시트르산 등의 성분이 들어 있어 심장을 강하게 하고 혈압을 내리며 면역력을 높여 주어 강장제로 쓰이며, 폐 기능을 강하게 하고 진해·거담 작용이 있어서 기침이나 갈증 등에 대하여 치료 효과가 있다. 오미자는 오래전부터 수렴, 자양, 강장, 진해약, 해주독, 목마름, 수렴고삽, 익기생진, 보신염심 등의 약효를 가져 생약원료로 한방에서 사용해오던 재료이며, 오미자주의 원료로 오래전부터 사용하여 체내 섭취에 따른 인체 안정성이 이미 확인되어 있다.Schisandra chinensis Baillon is a fruit of Schizandra chinensis Baillon tree, about 1cm in diameter, dark red. It has five flavors: sweet, sour, bitter, salty, and spicy, with the strongest of them. Schisandrarin, gomisin, citric acid, citric acid, citric acid and other ingredients, such as strong to strengthen the heart, lower blood pressure and increase immunity to use as a tonic, strong lung function and antitussive, expectorant action, cough or thirst It has a therapeutic effect against. Schisandra chinensis has long been used in herbal medicine as a raw material of medicinal herbs and has the effects of astringent, nourishment, tonic, ginhae medicine, haejudok, thirsty, convergent shovel, ripe ginjin, bosin dyed heart. Human stability with ingestion has already been confirmed.
어성초(Houttuynia Cordata)는 삼백초과에 속하는 다년생 초본으로, 한국백과사전에는 약모밀로 되어 있으며, 십약, 중약, 즙약 등으로 불리며 이외에 30여 가지의 이명이 있고, 별명으로 메밀나물이라고도 한다. 어성초는 항균작용이 있고, 항염, 진통, 조직재생 등의 효과가 있는 것으로 알려져 있고, 한방에서는 '피를 막게 하고 독을 없앤다'하여 청혈 해독제라 불리고 있다. 봄, 가을에 전초를 채취하여 세정한 후 말려서 청혈해독, 수종, 중풍, 폐렴, 맥대, 습진, 중이염 및 고혈압 치료에 쓰이며 강력한 항균효과를 지닌다. 실제로, katharrh 구균이나 influenza A형에 대한 항바이러스 작용이 우수하며, 또한 이뇨작용, 강심작용, 고혈압 예방 및 모세혈관 강화 작용이 매우 우수하다고 알려져 있다.Houttuynia Cordata is a perennial herb belonging to over three hundred and over, and is composed of medicinal herb in Korean encyclopedia. It is called as medicinal herb, Chinese herb, medicinal herb, and there are about 30 kinds of tinnitus. Eoseongcho has an antibacterial effect, and is known to have anti-inflammatory, analgesic, and tissue regeneration effects. In oriental medicine, it is called a blood detoxifying agent by 'clogging blood and eliminating poison'. It is used for the treatment of bleeding, decolonization, species, paralysis, pneumonia, eczema, eczema, otitis media and hypertension. It has strong antibacterial effect. In fact, it is known that antiviral activity against katharrh cocci and influenza type A is excellent, as well as diuretic, cardiovascular, hypertension and capillary strengthening.
상기 맥문동, 인삼, 오미자 및 어성초는 일반적으로 사용되는 건조물을 통째로 추출할 수도 있고, 적당한 크기로 잘라서(예컨대 길이 3 내지 20 ㎝, 오미자 제외) 추출하거나, 또는 분쇄하거나 분말로 갈아서 추출할 수 있다. 일반적으로 가장 효과적으로 유효성분을 추출하기 위해서는 적당히 부수어 추출하는 것이 바람직하다.The pulsed dong, ginseng, Schisandra chinensis and fish herb may be extracted as a whole, or may be extracted by cutting to a suitable size (for example, 3 to 20 cm in length, schisandra chinensis), or pulverized or ground. Generally, in order to extract the effective ingredient most effectively, it is preferable to break it and extract it.
상기 추출물의 추출방법으로는 온탕추출법, 열추출법, 환류냉각추출법, 초음파 추출법, 이산화탄소에 의한 초임계 추출법, 저온압착법, 일정한 분자량 컷-오프 값을 갖는 한외 여과막을 이용한 추출법, 다양한 크로마토그래피에 의한 추출법 등공지된 다양한 방법으로 추출할 수 있다.Extraction method of the extract is hot water extraction method, heat extraction method, reflux cooling extraction method, ultrasonic extraction method, supercritical extraction method using carbon dioxide, low temperature compression method, extraction method using an ultrafiltration membrane having a constant molecular weight cut-off value, by various chromatography It can be extracted by various known methods such as extraction method.
상기 추출물은 각 성분을 별도로 추출하여 혼합하거나, 상기 약재를 혼합한 후 추출하는 것에 의하여 제조될 수 있다. 즉 약재를 혼합하여 추출하는 것도 가능하고(혼합추출), 각각의 성분별로 추출한 후 추출물을 혼합하는 것도 가능하며, 약재를 혼합한 후 혼합 추출하는 것이 제조 효율이나 경제성 면에서 바람직하다. The extract may be prepared by separately extracting each component and mixing, or after mixing the medicinal herbs. That is, it is also possible to extract and mix the medicine (mixed extraction), it is also possible to mix the extract after extracting for each component, it is preferable in terms of manufacturing efficiency and economics to mix and extract the medicine.
본 발명에서 사용하는 추출물은 조추출액, 조추출액의 건조물, 조추출액의 농축물. 조추출액의 분획물, 분획물의 건조물 및 분획물의 농축물 중에서 선택된 하나 이상인 것을 특징으로 한다. 상기 언급한 형태에 국한되는 것은 아니나, 조추출액, 조추출액의 건조물, 조추출액의 농축물. 조추출액의 분획물, 분획물의 건조물 또는 분획물의 농축물이 피부 개선 효과가 우수하고, 추출물의 제조, 보관 및 취급과 제형 제조에 있어 편리성을 제공하여 바람직하다.Extract used in the present invention is crude extract, dried extract of crude extract, concentrate of crude extract. It is characterized in that at least one selected from fractions of crude extract, dried fractions and concentrates of fractions. Crude extracts, dried products of crude extracts, concentrates of crude extracts, but not limited to those mentioned above. Fractions of crude extracts, dried fractions or concentrates of fractions are preferred for their excellent skin-improving effect and for convenience in the preparation, storage and handling of extracts and formulation preparation.
본 발명에서 사용하는 추출물은 각 단계별로 얻어진 추출물을 모두 사용할 수 있지만, 어느 단계의 추출물을 본 발명에 따른 조성물에 적용하는가에 따라 소기의 효과를 달성하기 위한 함량이 달라진다. 예를 들어, 용매 추출 후 여과하여 바로 수득한 상기 조추출물을 사용하는 경우에는 조성물 총 중량 대비 0.05 내지 80.0 중량% 함유시키는 것이 바람직하다. 0.05 중량% 미만으로 함유시키면 본 발명이 목적하는 피부 개선 효과가 충분치 않을 수 있고, 이와 반대로 80.0 중량%를 초과하여 함유시키면 함량 증가에 따른 효과의 증가가 없어 비경제적일 뿐 아니라 제품의 안정성이 저하될 수 있다. 또한 감압농축장치와 동결건조기 등을 통해 추출물 내 유효성분의 함량을 선택적으로 증가시킨 건조물 또는 농축물은 경우 바람직한 사용함량은 조성물 총 중량 대비 0.001 내지 20.0 중량% 범위이다.The extract used in the present invention may use all of the extracts obtained in each step, but the content for achieving the desired effect varies depending on which step the extract is applied to the composition according to the present invention. For example, when using the crude extract directly obtained by filtration after solvent extraction, it is preferable to contain 0.05 to 80.0% by weight relative to the total weight of the composition. If the content is less than 0.05% by weight, the desired skin improvement effect of the present invention may not be sufficient. On the contrary, if the content is more than 80.0% by weight, the effect of increasing the content is not increased, which is not economical and the stability of the product is lowered. Can be. In addition, in the case of dried products or concentrates in which the content of the active ingredient in the extract is selectively increased through a reduced pressure concentrator and a lyophilizer, the preferred content is in the range of 0.001 to 20.0% by weight based on the total weight of the composition.
본 발명에서 사용하는 추출물은 당업계에 공지된 다양한 추출 방법을 이용하여 수득할 수 있으며, 용매에 의한 조추출물뿐만 아니라, 통상적인 정제 과정을 거친 추출물도 포함한다. 예컨대, 일정한 분자량 컷-오프 값을 갖는 한외여과막을 이용한 분리, 다양한 크로마토그래피(크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)에 의한 분리 등, 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획물도 본 발명의 추출물에 포함되는 것이다. 본 발명의 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.Extracts used in the present invention can be obtained using a variety of extraction methods known in the art, and includes not only crude extracts by a solvent, but also extracts obtained through conventional purification processes. Obtained by various additional purification methods, such as separation using ultrafiltration membranes having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity). Fractions are also included in the extract of the present invention. The extract of the present invention can be prepared in powder form by an additional process such as vacuum distillation and freeze-drying or spray-drying.
본 발명의 피부 개선용 약학 조성물의 피부 개선은 건조 피부의 보습력 향상, 아토피 피부염 증상 개선, 알레르기 증상 개선, 주름 개선, 피부톤 향상, 미백, 가려움증 개선, 피부 장벽기능 향상, 피부 면역기능 향상 및 염증 증상 개선 중 하나 이상인 것을 특징으로 한다.Improvement of the skin of the pharmaceutical composition for improving skin of the present invention improves the moisturizing power of dry skin, improves atopic dermatitis symptoms, improves allergic symptoms, improves wrinkles, improves skin tone, improves whitening, itching, improves skin barrier function, improves skin immune function and inflammation symptoms At least one of the improvements.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 맥문동, 인삼, 오미자 및 어성초의 추출물은 맥문동 : 인삼 : 오미자 : 어성초의 중량비율이 1 내지 8 : 1 내지 4 : 1 내지 4 : 1 내지 8 인 것(원료의 건조 중량 대비 기준)을 특징으로 한다. 맥문동, 인삼, 오미자 및 어성초의 혼합 비율에 제한되는 것은 아니나, 보다 우수한 피부 개선 효과를 나타내기 위해서는 상기 범위 내로 혼합하여 조성하는 것이 바람직하다. 그 중에서 맥문동 : 인삼 : 오미자 : 어성초의 중량비율을 1 : 1 : 1 : 2 로 혼합하는 경우(원료의 건조 중량 대비 기준)가 피부 개선 효과가 우수하여 가장 바람직하다. 상기 비율에 대응하는 범위 내라면, 각각의 추출물을 각기 대응하는 비율로 혼합하여 제조할 수도 있다.Extracts of McMoon-Dong, Ginseng, Schisandra chinensis and Eochochocho of the health functional food composition for improving skin diseases according to the present invention are the weight ratio of McMoon-Dong: Ginseng: Schisandra chinensis: Echochocho to 1-8: 1-4: 1-4: 1-8 It is characterized by (based on the dry weight of the raw material). Although it is not limited to the mixing ratio of ginmundong, ginseng, schisandra chinensis and fish herb, it is preferable to mix and composition in the said range in order to show the more excellent skin improvement effect. Among them, the weight ratio of Macmundong: Ginseng: Schisandra chinensis: Echoseongcho is 1: 1: 1: 2 (based on the dry weight of the raw material), which is most preferable because of excellent skin improvement effect. If it is in the range corresponding to the said ratio, each extract may be prepared by mixing each corresponding ratio.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 추출물은 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 한다. 원칙적으로 추출물의 용매의 종류에 제한되지 않으나, 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매가 바람직하며, 그 중에서도 정제수를 사용하여 추출한 추출물이 본 발명의 효과 및 제형 안정성 면에서 우수하여 바람직하다.The extract of the health functional food composition for improving skin diseases according to the present invention is characterized in that the extract is available as a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. In principle, the extract is not limited to the type of solvent, but a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof is preferable, and among these, an extract extracted using purified water is effective and formulation stability of the present invention. It is preferable because it is excellent in terms.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 추출물은 담체, 희석제, 부형제 및 첨가제 중 하나 이상을 더 포함하여 정제, 환제, 산제, 과립제, 분말제, 캡슐제 및 액제 제형으로 이루어진 군에서 선택된 하나로 제형된 것을 특징으로 한다. 본 발명의 추출물을 첨가할 수 있는 식품으로는, 각종 식품류, 분말, 과립, 정제, 캡슐, 시럽제, 음료, 껌, 차, 비타민 복합제, 건강기능성 식품류 등이 있다.Extract of the dietary supplement composition for improving skin diseases according to the present invention further comprises at least one of a carrier, a diluent, an excipient and an additive selected from the group consisting of tablets, pills, powders, granules, powders, capsules and liquid formulations. Characterized in one. Foods to which the extract of the present invention can be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gums, teas, vitamin complexes, and health functional foods.
상기 본 발명에 더 포함될 수 있는 첨가제로는, 천연 탄수화물, 향미제, 영양제, 비타민, 광물(전해질), 풍미제(합성 풍미제, 천연 풍미제 등), 착색제, 충진제(치즈, 초콜렛 등), 팩트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH조절제, 안정화제, 방부제, 산화 방지제, 글리세린, 알콜, 탄산화제 및 과육으로 이루어진 군으로부터 선택된 1종 이상의 성분을 사용할 수 있다. As additives that may be further included in the present invention, natural carbohydrates, flavors, nutrients, vitamins, minerals (electrolytes), flavors (synthetic flavors, natural flavors, etc.), colorants, fillers (cheese, chocolate, etc.), One or more components selected from the group consisting of fact acids and salts thereof, alginic acids and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonating agents and pulp .
상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상기 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
상기 외에 본 발명에 따른 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명에 따른 조성물은 천연 과일 쥬스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. In addition to the above, the composition according to the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the composition according to the present invention may contain fruit flesh for the production of natural fruit juices and vegetable drinks. These components can be used independently or in combination.
상기 담체, 부형제, 희석제 및 첨가제의 구체적인 예로는 이에 한정하는 것은 아니나, 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 에리스리톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 메틸셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일로 이루어진 그룹으로부터 선택된 1종 이상이 사용되는 것이 바람직하다.Specific examples of the carrier, excipient, diluent, and additives include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium phosphate, calcium Silicates, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate And at least one selected from the group consisting of mineral oils.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물을 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.When formulating a health functional food composition for skin disease improvement according to the present invention is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. that are commonly used.
상기 상술한 제형 내 유효성분으로서의 본 발명에 따른 추출물의 함량은 사용 형태 및 목적, 환자 상태, 증상의 종류 및 경중 등에 의하여 적절하게 조절할 수 있으며, 고형분 중량 기준으로 0.001 내지 99.9 중량%, 바람직하게는 0.01 내지 50 중량%일 수 있으나, 이에 한정되지 않는다.The content of the extract according to the present invention as an active ingredient in the above-described formulations can be appropriately adjusted according to the use form and purpose, patient condition, type of symptom and severity, etc. 0.01 to 50% by weight, but is not limited thereto.
본 발명의 추출물 함유 조성물의 투여 용량은 환자의 나이, 몸무게, 성별, 투여형태, 건강상태 및 질환 정도에 따라 달라질 수 있으며, 의사 또는 약사의 판단에 따라 일정 시간간격으로 1일 1회 내지 수회로 분할 투여할 수도 있다. 예컨대, 유효성분 함량을 기준으로 1일 투여량이 0.5 내지 500 ㎎/kg, 바람직하게는 1 내지 300㎎/kg일 수 있다. 상기한 투여량은 평균적인 경우를 예시한 것으로서 개인적인 차이에 따라 그 투여량이 높거나 낮을 수 있다. 본 발명의 혼합 추출물 함유 조성물의 1일 투여량이 상기 투여 용량 미만이면 유의성 있는 효과를 얻을 수 없을 수 있으며, 그 이상을 초과하는 경우 비경제적일 뿐만 아니라 상용량의 범위를 벗어나므로 바람직하지 않은 부작용이 나타날 수 있다.The dosage of the extract-containing composition of the present invention may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and once or several times a day at regular intervals according to the judgment of the doctor or pharmacist. It may be administered in divided doses. For example, the daily dosage may be 0.5 to 500 mg / kg, preferably 1 to 300 mg / kg, based on the active ingredient content. The above dosages are illustrative of the average case and may be high or low depending on individual differences. If the daily dose of the mixed extract-containing composition of the present invention is less than the dosage, it may not be possible to obtain a significant effect. Can be.
본 발명에 따르면, 맥문동, 인삼, 오미자 및 어성초에 용매를 가하여 추출하여 추출물을 수득하는 단계; 및 상기 추출물을 농축하는 단계를 포함하는 피부 질환 개선용 건강기능식품 조성물의 제조방법을 제공한다. 상기 추출하는 방법은 통상의 방법으로 수행될 수 있고, 상기 추출물에서 상술한 바와 같다.According to the present invention, a step of extracting by adding a solvent to the ganmundong, ginseng, Schisandra chinensis and Eochocho to obtain an extract; And it provides a method for producing a health functional food composition for improving skin diseases comprising the step of concentrating the extract. The extraction method may be performed by a conventional method, as described above in the extract.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 제조방법의 추출물을 수득하는 단계는 맥문동, 인삼, 오미자 및 어성초의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출되는 것을 특징으로 한다. 비록 상기 용매의 양에 제한되는 것은 아니나, 상기보다 용매의 양이 더욱 많은 경우에는 유효 성분의 농도가 낮거나 추출에 드는 에너지나 시간이 많이 소모되고, 이보다 더 적은 경우에는 초기 재료 투입량 대비 최종 추출물의 수율이 감소할 수 있다.Obtaining the extract of the method for producing a health functional food composition for improving skin diseases according to the present invention is characterized in that the extract by adding a solvent of 5 to 20 times the weight of the total weight of ganmundong, ginseng, Schisandra chinensis and eoseongcho. Although not limited to the amount of the solvent, when the amount of the solvent is higher than the above, the concentration of the active ingredient is low or the energy or time required for extraction, and if less than the final extract compared to the initial material input Yield can be reduced.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 제조방법의 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 수행되는 것을 특징으로 한다. 비록 상기 시간과 온도에 국한되는 것은 아니나 유효 성분의 실질적인 용출을 위해서는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 수행되는 것이 바람직하며, 그 중에서도 95℃ 내지 115℃에서 3시간 내지 5시간 동안 수행되는 것이 유효 성분의 용출면에서 가장 효과적이다. 상기 온도보다 더 낮거나 시간이 짧은 경우에는 유효성분의 실질적인 용출이 감소하고, 상기 온도보다 더 높거나 시간이 긴 경우에는 추출 공정의 시간이나 효율이 저하되거나 유효성분이 파괴되거나 바람직하지 않은 성분이 혼합되거나 맛이 변할 우려도 있다. Obtaining the extract of the method for producing a health functional food composition for improving skin diseases according to the present invention is characterized in that it is carried out at 80 ℃ to 120 ℃ for 1 hour to 10 hours. Although not limited to the above time and temperature, it is preferable to perform 1 hour to 10 hours at 80 ° C. to 120 ° C. for substantial dissolution of the active ingredient, and particularly, 3 hours to 5 hours at 95 ° C. to 115 ° C. It is most effective in terms of dissolution of the active ingredient. If the temperature is lower or shorter than the temperature, the actual elution of the active ingredient is reduced. If the temperature is higher or longer than the temperature, the time or efficiency of the extraction process is reduced, the active ingredient is destroyed, or undesirable components are mixed. Or taste may change.
본 발명에 따른 피부 질환 개선용 건강기능식품 조성물의 제조방법의 농축하는 단계는 보다 구체적으로 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화하는 단계를 포함하는 것을 특징으로 한다.Concentrating the manufacturing method of the health functional food composition for improving skin diseases according to the present invention more specifically the step of filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; And pulverizing the dried concentrate.
상기 추출물을 여과, 농축, 건조, 분쇄하는 방법은 추출물에 대하여 통상적으로 적용할 수 있는 여과, 농축, 건조, 분쇄방법이면 특정한 방법에 국한되지 않는다. 보다 바람직하게는 여과과정에서는 추출된 유효성분이 여과액 속에 포함될 수 있으면서 불순물은 제거할 수 있는 적당한 크기의 필터를 사용하고, 여과에서 분쇄의 일련의 과정은 유효성분의 파괴를 막기 위하여 60℃이하에서 진행하는 것이 바람직하다.The method of filtering, concentrating, drying and pulverizing the extract is not limited to a specific method as long as it is a filtration, concentration, drying and pulverization method that can be commonly applied to the extract. More preferably, in the filtration process, a filter of a suitable size capable of removing impurities while containing the extracted active ingredient may be included in the filtrate, and a series of pulverization in filtration may be performed at 60 ° C. or lower to prevent destruction of the active ingredient. It is preferable to proceed.
따라서, 본 발명에 사용되는 추출물은 바람직하게는 하기와 같은 방법으로 수득될 수 있다. 먼저, 맥문동, 인삼, 오미자 및 어성초 각각 분량에 물을 일정량 첨가한 후 환류장치 하에서 약 1시간 내지 10시간 동안 끓이면서 추출한 후 여과하여 그 여액을 취하고 물을 첨가하여 총량을 조정한 다음, 저온에서 1일 내지 12일간 방치하여 생성된 침전물들을 막 여과를 실시함으로써 추출물을 수득할 수 있다.Therefore, the extract used in the present invention can preferably be obtained by the following method. First, add a certain amount of water to each of the mungmundong, ginseng, Schisandra chinensis and Eoseongcho and then extract it while boiling for about 1 hour to 10 hours under reflux, and then filtrate the filtrate and adjust the total amount by adding water, and then at 1 low temperature. The precipitate can be obtained by performing membrane filtration on the resulting precipitate left for 1 to 12 days.
또한 생약재 내 존재하는 유효성분의 충분한 추출을 위하며, 상기 과정에 의해 1회 추출한 생약재에 다시 한번 용매를 가하여 다시 한번 추출하는 과정을 부가할 수 있다. 이 경우 첫 번째 추출한 추출물과 두 번째 추출한 추출물을 혼합하여 동시에 여과 및 농축과정을 거친다면 더욱 효율적이고 바람직하다.
In addition, for the sufficient extraction of the active ingredient present in the herbal medicine, by adding the solvent once again to the herbal medicine extracted once by the above process may be added to the extraction process again. In this case, it is more efficient and preferable if the first and second extracts are mixed and filtered and concentrated at the same time.
이하에서는 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 다만, 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다 할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.
본 발명에 따른 실시예 1은 다음과 같은 과정을 통해 제조하였다. Example 1 according to the present invention was prepared through the following process.
맥문동, 인삼, 오미자 및 어성초를 시중에서 구입하여 맥문동 40g, 인삼 40g, 오미자 40g 및 어성초 80g을 추출기에 넣은 다음, 정제수를 2.7L 가하고 112℃ 에서 3시간 가온 추출하여 추출액 1.5L 를 수득하였다.Megmundong, ginseng, Schisandra chinensis and Eochocho were purchased on the market, 40g of Macmundong, 40g of Ginseng, 40g of Schisandra chinensis and 80g of Echochocho were added to an extractor, 2.7L of purified water was added thereto, and extracted by heating at 112 ° C for 3 hours to obtain 1.5L of extract.
비교예는 어성초를 첨가하지 않은 것을 제외하고는 상기 실시예 1의 제조와 동일한 방법으로 제조하였다.Comparative Example was prepared in the same manner as in Preparation of Example 1, except that the fish paste was not added.
고형분 측정기로 전탕액의 농도를 측정한 결과, 실시예 1 추출액의 농도는 10.3%, 비교예 추출액의 농도는 9.8% 였다.As a result of measuring the density | concentration of the hot water liquid with a solid content measuring device, the density | concentration of the extract of Example 1 was 10.3%, and the density | concentration of the extract of the comparative example was 9.8%.
상기에서 얻어진 각각의 추출물을 여과포를 사용하여 여과하고 동결건조기를 이용하여 냉동 건조한 다음, 이를 4℃ 에서 보관하여 사용하였다.
Each extract obtained above was filtered using a filter cloth and lyophilized using a lyophilizer, which was then stored and used at 4 ° C.
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시험예Test Example
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1. 실험의 준비 및 실험 방법1. Preparation and Experiment Method
1) 실험동물의 준비1) Preparation of experimental animals
웅성(雄性) 8주령의 NC/Nga atopy mice를 중앙실험동물에서 공급받아 실험당일까지 고형사료와 물을 공급하고 실온 22±2℃, 상대습도 50~55%, 조도 200 lux(8시 점등, 20시 소등)를 계속 유지하면서 1주일간 실험실 환경에 적응시킨 후 실험에 사용하였다.
8 week-old NC / Nga atopy mice were fed from a central laboratory animal and fed solid feed and water until the day of the experiment. Room temperature 22 ± 2 ℃, relative humidity 50-55%,
2) 시약의 준비2) Preparation of Reagents
본 실험에 사용된 시약으로 디에틸피로카보네이트(diethyl pyrocarbonate, DEPC), 염화암모늄(NH4Cl), 탄산수소칼륨(KHCO3), 에틸렌디아민테트라아세트산화 나트륨(Na2EDTA), 완전보조제(complete adjuvant), 클로로포름(chloroform), 콜라게나제(collagenase) IV, 배지 RPMI-1640, 이소프로판올(isopropanol), 적혈구용혈액(ACK lysis solution), 브롬화 에티듐(ethidium bromide, EtBr), 둘베코즈 인산완충식염수(Dulbecco's phosphate buffered saline, D-PBS), 디엠이엠 배지(Dulbecco's minimum essential medium, DMEM), 포름알데하이드(formaldehyde), 염화마그네슘(magnesium chloride, MgCl2) 및 아가로스(agarose)는 Sigma 사(USA) 제품을, 우태아혈청(fetal bovine serum, FBS)은 Gibco BRL사(USA) 제품을, RNase 저해제(RNase inhibitor), Taq 중합효소(Taq polymerase), 마구잡이시발체(random primer), 디옥시리보뉴클레오타이드(Deoxynucleotide triphosphate, dNTP) 및 Moloney murine leukemia virus reverse transcriptase(M-MLV RT)는 Promega 사(USA) 제품을, 인터루킨-4(IL-4), 인터루킨 5(IL-5), 인터루킨 6(IL-6) 및 인터페론감마(IFN-γ)의 ELISA kit는 BD(USA)사 제품을, 면역글로블린 E(IgE), 면역글로블린M(IgM) 및 면역글로블린G1(IgG1)의 ELISA kit는 고마바이오텍(Korea) 제품을, RNAzolB는 invitrogen 사(USA) 제품을 사용하였고, 그 외 사용된 시약은 Duksan(Korea)에서 구입하여 사용하였다.
The reagents used in this experiment were diethyl pyrocarbonate (DEPC), ammonium chloride (NH 4 Cl), potassium bicarbonate (KHCO 3 ), ethylenediaminetetraacetate (Na 2 EDTA), and complete adjuvant (complete). adjuvant, chloroform, collagenase IV, medium RPMI-1640, isopropanol, erythrocyte ACK lysis solution, ethidium bromide (EtBr), Dulbecco's phosphate buffered saline (Dulbecco's phosphate buffered saline (D-PBS), Dulbecco's minimum essential medium (DMEM), formaldehyde, magnesium chloride (magnesium chloride, MgCl 2 ) and agarose are available from Sigma Corporation (USA). Products, fetal bovine serum (FBS) is a Gibco BRL (USA) product, RNase inhibitor, RNase inhibitor, Taq polymerase, random primer, deoxynucleotide triphosphate , dNTP) and Moloney murine leukemia virus reverse transcriptase (M-MLV RT) uses Promega (USA) products such as interleukin-4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6) and interferongamma (IFN- γ) ELISA kit from BD (USA), immunoglobulin E (IgE), immunoglobulin M (IgM) and immunoglobulin G1 (IgG1) ELISA kit of Goma Biotech (Korea), RNAzolB is invitrogen (USA) was used, and other used reagents were purchased from Duksan (Korea).
3) 인간 각질 형성 세포주 배양, 3) culturing human keratinocyte cell lines, 실시예와Examples and 비교예의Comparative example 처리 process
실험에 사용한 인간 각질 형성 세포주 (Human kerationocyte HaCaT cell line)는 DMEM 배지에 10% fetal bovine serum(FBS; Gibco CO., USA), 1% 페니실린-스테렙토마이신(penicilin-streptomycin, Gibco Co., USA) 을 첨가하여 37℃, 5% CO2 조건하에서 배양하여 사용하였다. 세포를 75mm plate에 약 80%의 confluency에 도달할 때까지 배양하였다. Human kerationocyte HaCaT cell line used in the experiment was prepared in 10% fetal bovine serum (FBS; Gibco CO., USA), 1% penicilin-streptomycin (Gibco Co., USA) in DMEM medium. ) Was used to incubate at 37 ℃, 5% CO 2 conditions. Cells were incubated in a 75 mm plate until reaching a confluency of about 80%.
상기에서 마련한 비교예 및 실시예 1을 각각 증류수에 희석하여 1%(w/v)의 stock을 만들어, 배지 내 최종 농도가 0.01, 0.02, 0.05, 0.1%가 되도록 처리하여, CO2 배양기에서 24시간 동안 배양하였다. 대조군은 상기에서 비교예 및 실시예 1을 첨가하지 않은 배지에서 배양한 것을 제외하고는 동일한 방법을 사용하였다.
Diluted in the Comparative Example and Example 1 provided in the respective distilled water to make a stock of 1% (w / v), the processing is in the final concentration of the medium to be 0.01, 0.02, 0.05, 0.1% in the CO 2 incubator 24 Incubated for hours. As a control, the same method was used except that the control was incubated in a medium without addition of Comparative Example and Example 1.
4) 세포 생존능력 시험(4) Cell viability test CellCell viabilityviability testtest ))
세포 생존능력 시험은 cell counting assay를 이용하여 진행하였다. 상기에서 제조한 추출물 시료(실시예 1 및 비교예)를 각각 농도별로 처리하여 24시간 배양 후 처리된 배지와 시약을 제거하였다. 200㎕의 트립신-이디티에이 완충액(Trypsin-EDTA buffer, TE buffer)를 처리한 후 incubator에서 2~3분 동안 방치하였다. 1.8㎖의 인산완충식염수를 첨가한 후 세포를 plate에서 떨어뜨린 후 10㎕의 트리판 블루(Trypan blue)와 10㎕의 세포액을 잘 섞은 후 실온에서 3~4분 동안 반응시켰다. Cell viability test was conducted using a cell counting assay. The extract samples (Example 1 and Comparative Example) prepared above were treated for each concentration to remove the treated medium and reagents after 24 hours of incubation. After treatment with 200 μl of trypsin-EDTA buffer (Trypsin-EDTA buffer, TE buffer) and left in the incubator for 2-3 minutes. After adding 1.8 ml of phosphate buffered saline, the cells were dropped from the plate, 10 μl of Trypan blue and 10 μl of cell solution were mixed well, and reacted at room temperature for 3 to 4 minutes.
혈구계수기(Hemacytometer)에 덮개 유리(cover glass)를 올려놓은 후 hemacytometer와 cover glass사이의 틈에 반응물을 10㎕ 취하여 주입한 후 현미경을 이용하여 염색되지 않은 세포와 염색된 세포의 수를 측정하였다.
After the cover glass was placed on the hemacytometer, 10 µl of the reactant was injected into the gap between the hemacytometer and the cover glass, and the number of unstained and stained cells was measured using a microscope.
5) 5) 타이로시네이즈Tairosineiz 저해 활성 ( Inhibitory activity ( TyrosinaseTyrosinase inhibitioninhibition activityactivity ) 분석) analysis
각 조건별 시료를 처리한 세포 추출물을 취한 다음, 96 well plate에 세포 추출물 100㎕와 500U tyrosinase 20㎕ 및 1.5mM L-DOPA 80㎕(A)를 각각 가하고 37℃에서 10분간 반응시킨 후 450nm에서 흡광도를 측정하였다. 그리고 tyrosinase 대신 인산완충식염수(PBS)를 참가하여 동일한 방법으로 측정한 Blank 값(B)과 시료용액 대신 lysis buffer를 첨가하고 위와 같이 tyrosinase를 첨가(C)한 것과 PBS를 첨가(D)하여 동일한 방법으로 37℃에서 10분간 반응시킨 후 450nm에서 흡광도를 측정한 값으로부터 하기 수학식 1에 의해 타이로시네이즈에 대한 저해 활성을 계산하였다.
After taking the cell extract treated with each sample, 100 μl of cell extract, 20 μl of 500U tyrosinase and 80 μl of 1.5 mM L-DOPA were added to a 96 well plate and reacted at 37 ° C. for 10 minutes, and then at 450 nm. Absorbance was measured. And phosphate buffered saline (PBS) instead of tyrosinase was added in the same way by measuring the blank value (B) and lysis buffer instead of the sample solution and tyrosinase (C) and PBS (D) as above. After the reaction at 37 ° C. for 10 minutes, the inhibitory activity against tyrosinase was calculated by the following
6) 6) 필라그린Pillar Green (( FilaggrinFilaggrin ) 과 ) And 세린팔미토일Serine palmitoyl 트랜스퍼레이즈( Transferraise ( serineserine palmitoylpalmitoyl transferase, transferase, SPTSPT ) 발현 분석) Expression analysis
필라그린(Filaggrin)의 발현을 평가하기 위하여, 상기의 방법으로 배양 및 시험한 세포로부터 트리졸(Trizol, invitrogen CO., USA)을 이용하여 total RNA를 추출한 다음, one step RNA PCR kit(AMV) (Takara Bio Inc., Japan)에서 제공되는 방법에 준하여 역전사 중합효소 연쇄반응(reverse transcription polymerase chain reaction, 이하 RT-PCR) 분석을 시행하였다. PCR 증폭은 GeneAmp® PCR System 2700(Applied Biosystems, USA)을 이용하여 94℃에서 30초 동안 열변성(denaturation) 시킨 후, 60℃에서 30초 동안 서냉복원(annealing)을 하고, 72℃에서 75초 동안 DNA 중합효소에 의한 확장(extension)이 되도록 하였다. 상기 반응에서 얻어지는 증폭 산물은 필라그린(filaggrin)과 β-액틴(actin)이 각기 약 400bp와 300bp이다. 이때, filaggrin에 대한 프라이머는 바이오니아사(Korea)의 5'-GGTAGATAGATCTGGACACTCAGGG-3'를 사용하였으며, internal control으로는 β-액틴을 이용하여 발현량을 비교하였다. To evaluate the expression of Filaggrin, total RNA was extracted from the cells cultured and tested by the above method using Trizol, invitrogen CO., USA, and then one step RNA PCR kit (AMV). According to the method provided by (Takara Bio Inc., Japan), reverse transcription polymerase chain reaction (RT-PCR) analysis was performed. PCR amplification was performed using GeneAmp ® PCR System 2700 (Applied Biosystems, USA) for 30 seconds of denaturation at 94 ° C, followed by slow cooling at 60 ° C for 30 seconds, and 75 ° C at 72 ° C. During extension by DNA polymerase. The amplification products obtained in the reaction are filaggrin (filaggrin) and β-actin (about 400bp and 300bp, respectively). At this time, the primer for filaggrin was used 5'-GGTAGATAGATCTGGACACTCAGGG-3 'of Bioneer Corporation (Korea), and the expression level was compared using β-actin as the internal control.
세린팔미토일 트랜스퍼레이즈(serine palmitoyl transferase, SPT) 발현을 분석하기 위하여, SPT의 mRNA 수준을 Dorfman과 Lichtenstein의 방법에 따라 RT-PCR로 분석하였다. 상기의 방법으로 배양 및 시험한 세포로부터 트리졸을 이용하여 total RNA를 추출하고, First strand cDNA synthesis kit (Fermentas, USA)에서 제공되는 방법에 준하여 cDNA를 합성하였다. RT-PCR은 1㎕ template, 10㎕ PCR master mix(Qiagen, USA), 1㎕ β-actin과 7㎕ 증류수로 총 부피를 20㎕로 한 PCR mix로 PCR을 수행하였다. 상기 PCR 수행 조건은 denature (95℃, 15초), annealing, extension(60℃, 60초)으로 45 cycle을 수행하였으며, mRNA의 양은 comparative CT method를 이용하여 각각의 template의 변이에 기인하는 증폭양을 normalization하였고, 또한 house keeping gene인 β-actin을 각 유전자의 기준으로 정하여 정량을 유도하였고, 대조시료에서의 각 유전자를 표준기(calibrator)로 정하여 정량을 하였다.
To analyze serine palmitoyl transferase (SPT) expression, mRNA levels of SPT were analyzed by RT-PCR according to Dorfman and Lichtenstein's method. Total RNA was extracted from the cells cultured and tested by the above method using trizol, and cDNA was synthesized according to the method provided by First strand cDNA synthesis kit (Fermentas, USA). RT-PCR was performed by PCR mix with a total volume of 20 μl with 1 μl template, 10 μl PCR master mix (Qiagen, USA), 1 μl β-actin and 7 μl distilled water. The PCR conditions were 45 cycles with denature (95 ℃, 15 seconds), annealing, extension (60 ℃, 60 seconds), and the amount of mRNA was amplified due to variation of each template using comparative CT method. In addition, the house keeping gene β-actin was determined as the reference for each gene, and quantification was induced, and each gene in the control sample was determined as a calibrator.
7) 단백질 추출 및 7) protein extraction and COXCOX -2, -2, APAP -1의 -1 of WesternWestern 분석 analysis
COX-2와 AP-1의 발현을 평가하기 위하여 상기의 방법으로 배양 및 시험한 세포에 라이시스 버퍼(lysis buffer)를 처리하여 단백질을 추출하였다. 상기 라이시스 버퍼는 다음과 같은 조성으로 제조하였다; 0.1% 트리톤(Triton) X-100, 20mM Tris-Cl(pH7.4), 1mM 에틸렌디아민사아세트산(ethylenediaminetetraacetic acid, EDTA), 1mM 에틸렌글리콜비스(2-아미노에틸에테르)4-아세트산(ethylene glycol bis(2-aminoethyl ether) tetraacetic acid, EGTA), 150mM 염화나트륨(NaCl). In order to evaluate the expression of COX-2 and AP-1, the cells were cultured and tested by the above method, and the protein was extracted by treating lysis buffer. The Lysis buffer was prepared in the following composition; 0.1% Triton X-100, 20 mM Tris-Cl (pH 7.4), 1 mM ethylenediaminetetraacetic acid (EDTA), 1 mM ethylene glycol bis (2-aminoethylether) 4-acetic acid (ethylene glycol bis (2-aminoethyl ether) tetraacetic acid (EGTA), 150 mM sodium chloride (NaCl).
30㎍의 단백질을 10% SDS-폴리아크릴아마이드 젤(polyacrylamide gel)에 전기영동한 후, 니트로셀룰로오스 멤브레인(nitrocellulose membrane)으로 단백질을 이동시키고, COX-2, AP-1, Actin 부분의 membrane을 blocking solution(탈지분유를 1×TBS-T 용액에 5 %로 희석)으로 1시간 동안 blocking 시킨다. 1차 항체를 4℃에서 12시간 동안 반응시키고, 세척 후 2차 항체를 반응시킨 후 ECL western blotting detection system을 이용하였다.
After electrophoresis of 30 µg of protein on a 10% SDS-polyacrylamide gel, the protein was transferred to a nitrocellulose membrane, and the membrane of COX-2, AP-1, and Actin was blocked. Block for 1 hour with solution (diluted skim milk powder in 5% in 1 × TBS-T solution). The primary antibody was reacted at 4 ° C. for 12 hours, and after washing, the secondary antibody was reacted with an ECL western blotting detection system.
8) 염증유발 실험8) Inflammatory Experiment
NC/Nga mice는 실험군별로 각각 6마리를 실험하였으며 각 케이지별로 3마리씩 사육하였다. 즉 물을 제공한 대조군 6마리, 비교예를 제공한 비교군 6마리, 및 실시예 1를 제공한 실험군 6마리로, 총 NC/Nga mice 18마리에 cage 6개로 나누어 실험을 실시하였다.NC / Nga mice were tested for 6 animals per experimental group and 3 animals for each cage. That is, the experiment was carried out by dividing into six cages in 18 NC / Nga mice and 6 control groups provided with water, 6 comparative groups provided with a comparative example, and 6 experimental groups provided with Example 1.
상기 실험 동물 모두에 1-클로로-2,4-디니트로벤젠(1-chloro 2,4-dinitrobenzene, DNCB)을 이용하여 아토피 피부염을 유발하였다. 먼저 NC/Nga mice의 등 부위를 깨끗이 제모한 후 피부의 미세 상처가 치유되도록 24시간 방치하였다. 실험 4일 전 1% DNCB 용액(acetone:olive oil=3:1) 200㎕를 등 부위에 도포하고, 실험 시작일부터 종료일까지 4주간 일주일에 2번씩 0.2% DNCB용액 150㎕를 등 부위에 도포하여 염증을 유발하였다.Atopic dermatitis was induced in all of the experimental animals using 1-chloro-2,4-dinitrobenzene (DNCB). First, the back of the NC / Nga mice was epilated and left for 24 hours to heal fine wounds on the skin. 200 μl of 1% DNCB solution (acetone: olive oil = 3: 1) was applied to the
아토피 피부염이 유발된 쥐에게 4주 동안 실험을 진행하였고, 그 기간 동안 대조군은 음용수만을 투여하였고, 비교군은 어성초를 함유하지 않은 비교예를 혼합한 음용수를, 실험군은 실시예 1을 혼합한 음용수를 제공하였다. 상기 비교군 및 실험군의 음용수는 증류수 150g에 추출물 150㎎을 첨가(0.1% 희석)하여 제공하였으며, 이는 실험 동물 한 마리당 추출물 50㎎을 공급하는 양에 해당한다.
Experiments were conducted in rats with atopic dermatitis for 4 weeks, during which time the control group was administered only drinking water, and the comparison group was drinking water mixed with a comparative example containing no fish herb, and the experimental group was mixed with Example 1 Provided. Drinking water of the comparison group and the experimental group was provided by adding 150 mg of extract (0.1% dilution) to 150 g of distilled water, which corresponds to the amount of 50 mg of extract per animal.
9) 피부 손상도 측정9) Skin damage measurement
NC/Nga mice의 피부 손상도는 아토피성 피부염에서 일반적으로 사용되는 임상적 육안 평가법을 이용하였다. 육안평가 결과는 홍반(erythema), 가려움과 건조피부(Pruritus & Dry Skin), 부종과 혈종(Edema & escoriation), 미란(Erosion), 및 태선화(Lichenification)의 5가지 항목을 각각 평가한 점수의 총합으로 나타내었다. 이 각각의 항목은 없음(0), 약함(1), 중증도(2), 심함(3)으로 나누어서 채점하였다.
The skin damage of NC / Nga mice was evaluated by clinical visual evaluation method commonly used in atopic dermatitis. The gross evaluation results are the sum of the scores of each of the five categories of erythema, itching and dry skin, edema and escoriation, erosion, and lichenification. As shown. Each of these items was scored by dividing into none (0), weak (1), severity (2), and severe (3).
11) 11) 혈청내In serum 인터루킨( Interleukin ( ILIL )-4,5,6 및) -4,5,6 and 면역글로불린M(Immunoglobulin M ( IgMIgM ), 면역글로불린G1(IgG1), ), Immunoglobulin G1 (IgG1), 인터페론감마Interferongamma (( IFNIFN -γ) 정량γ) quantification
추출물 투여 4주 후 NC/Nga 생쥐를 에틸에테르(ethyl ether)로 마취한 후 심장 천자법으로 혈액을 채혈하여 혈청을 분리한 후 혈청 중 인터루킨(interlukin, IL)-4, 5, 6 및 면역글로불린M(immunoglobulin M, IgM), 면역글로불린G1(immunoglobulin G1, IgG1), 인터페론감마(interferon-γ, IFN-γ) 양을 각각 ELISA kit로 정량하였다.
Four weeks after the administration of the extract, NC / Nga mice were anesthetized with ethyl ether, and blood was collected by cardiac puncture to separate the serum, and then interlukin (IL) -4, 5, 6 and immunoglobulin in serum. The amount of M (immunoglobulin M, IgM), immunoglobulin G1 (immunoglobulin G1, IgG1), and interferon-gamma (interferon-γ, IFN-γ) were respectively quantified by ELISA kit.
2. 추출물이 세포에 미치는 영향2. Effect of Extracts on Cells
1) 세포 성장에 미치는 영향1) Effect on Cell Growth
본 발명에 따른 추출물이 피부 세포에 미치는 영향을 평가하였다. 실시예 1과 비교예를 각각 0, 0.01, 0.02, 0.05, 0.1%의 농도로 배지에 첨가하여 배양하였을 때의 결과를 도 1에 나타내었다.The effect of the extract according to the invention on the skin cells was evaluated. The results obtained when Example 1 and Comparative Example were added to the culture medium at concentrations of 0, 0.01, 0.02, 0.05 and 0.1%, respectively, are shown in FIG. 1.
도 1에 나타나 있는 것처럼, 두 가지 경우 모두 세포 생존에 유해한 영향을 주지 않았고, 세포 성장도 대조군(대조군의 배양 사진은 미도시)과 같이 아무런 영향없이 잘 이루어지는 것을 확인할 수 있었다.
As shown in Figure 1, both cases did not have a detrimental effect on cell survival, it was confirmed that the cell growth is well performed without any effect as a control (culture picture of the control group not shown).
2) 2) 타이로시네이즈Tairosineiz (( tyrosinasetyrosinase ) 억제 활성에 미치는 영향) Effect on inhibitory activity
추출물이 tyrosinase 억제 활성에 미치는 영향을 알아보기 위해서 비교예 및 실시예 1을 인간각질형성세포 HaCaT cell에 각각 0.01, 0.02, 0.05, 0.1% 농도별로 배양배지에 처리하였으며, 그 결과는 도 2에 나타내었다. (a)는 비교예를 처리한 실험 결과이고, (b)는 실시예 1을 처리한 실험 결과이다.In order to examine the effect of the extract on tyrosinase inhibitory activity, Comparative Example and Example 1 were treated in culture media by 0.01, 0.02, 0.05, and 0.1% concentrations in human keratinocyte HaCaT cells, respectively, and the results are shown in FIG. 2. It was. (a) is the experimental result which processed the comparative example, (b) is the experimental result which processed Example 1.
도 2에 나타난 것처럼, 비교예(a)에서는 농도가 증가함에 따라 tyrosinase 억제활성이 감소하는 경향을 보였으나, 반대로 실시예 1(b)에서는 농도 의존적으로 tyrosinase 억제 활성이 증가함을 보였다. As shown in FIG. 2, in the comparative example (a), the tyrosinase inhibitory activity tended to decrease as the concentration was increased. In contrast, in Example 1 (b), the tyrosinase inhibitory activity was increased in a concentration-dependent manner.
즉, 실시예 1에 의해 tyrosinase 활성이 억제되면서 멜라닌과 그 밖에 다른 색소들의 생성을 억제시켜 피부 노화시 생성될 수 있는 반점 등의 생성을 억제시킬 수 있다는 것을 알 수 있다.
In other words, it can be seen that by inhibiting the production of melanin and other pigments while inhibiting tyrosinase activity according to Example 1, it is possible to inhibit the production of spots and the like that may be generated during skin aging.
3) 피부 장벽 조절 인자에 미치는 영향3) Effects on Skin Barrier Regulators
추출물이 피부 장벽 기능 조절인자에 미치는 영향을 알아보기 위해서, 비교예와 실시예 1을 각각 0.01, 0.02, 0.05, 0.1% 농도별로 처리하여 필라그린(filaggrin)과 세린팔미토일 트랜스퍼레이즈(SPT)의 mRNA 발현을 reverse transcription(RT) PCR로 분석하였으며, 그 결과를 도 3 및 도 4에 나타내었다. (a)는 비교예를 처리한 실험 결과이고, (b)는 실시예 1을 처리한 실험 결과이다.In order to examine the effect of the extract on skin barrier function regulators, Comparative Example and Example 1 were treated by 0.01, 0.02, 0.05 and 0.1% concentrations of filaggrin and serine palmitoyl transferase (SPT), respectively. mRNA expression was analyzed by reverse transcription (RT) PCR, and the results are shown in FIGS. 3 and 4. (a) is the experimental result which processed the comparative example, (b) is the experimental result which processed Example 1.
도 3 및 도 4에 나타난 것처럼, 비교예(a)에서는 농도 의존적으로 filaggrin의 발현양이 감소하였으나, 실시예 1(b)에서는 filaggrin 발현양이 농도 의존적으로 증가하다가 0.05%에서 가장 높은 발현양을 보였다. 또한 SPT의 발현양 역시 비교예(a)에서는 0.01%에서 잠시 증가하는 듯 하였으나 농도 의존적으로 감소하는 양상을 보였고, 실시예 1(b)에서는 농도의존적으로 증가하며 0.05%에서 가장 높은 발현양을 보였다. 3 and 4, in Comparative Example (a), the expression level of filaggrin was decreased in a concentration-dependent manner. In Example 1 (b), the expression level of filaggrin was increased in a concentration-dependent manner. Seemed. In addition, the expression level of SPT also appeared to increase temporarily at 0.01% in Comparative Example (a), but showed a decrease in concentration-dependent manner. In Example 1 (b), the expression level increased in concentration and showed the highest expression level at 0.05%. .
이를 종합하면 비교예는 피부 장벽 조절 인자의 발현에 좋지 않은 영향을 미치는 것으로 보아 그에 노출되면 피부에 자극을 주는 것으로 판단되지만, 실시예 1은 피부 장벽 조절 인자의 발현에는 좋은 영향을 미치는 것을 확인할 수 있었다. 따라서 실시예 1은 피부 장벽 조절인자인 filaggrin과 SPT의 양을 증가시켜 줌으로써 피부의 수분 유지 기능을 유지시켜주는 기능을 강화시키는 것을 알 수 있다.
Taken together, the comparative example was found to adversely affect the expression of the skin barrier regulator, and when exposed to it, it is judged to be irritating to the skin, but in Example 1, it can be seen that it has a good effect on the expression of the skin barrier regulator. there was. Therefore, it can be seen that Example 1 enhances the function of maintaining the skin's water retention function by increasing the amount of skin barrier regulators, filaggrin and SPT.
4) 피부 항염증 반응 인자에 미치는 영향4) Effect on skin anti-inflammatory response factors
추출물이 피부 항염증 반응 인자에 미치는 영향을 알아보기 위해서, 비교예와 실시예 1을 각각 0.01, 0.02, 0.05, 0.1% 농도별로 처리하여 COX-2의 발현과 COX-2의 전사인자(transcription factor)중 하나인 AP-1의 발현을 western blotting으로 분석하였으며, 그 결과를 도 5 및 도 6에 나타내었다. (a)는 비교예를 처리한 실험 결과이고, (b)는 실시예 1을 처리한 실험 결과이다.To investigate the effect of the extract on skin anti-inflammatory response factors, Comparative Example and Example 1 were treated by 0.01, 0.02, 0.05 and 0.1% concentrations, respectively, to express COX-2 and transcription factor of COX-2. Expression of AP-1 was analyzed by western blotting, and the results are shown in FIGS. 5 and 6. (a) is the experimental result which processed the comparative example, (b) is the experimental result which processed Example 1.
도 5 및 도 6에 나타난 것처럼, 비교예(a)에서는 0.02% 처리한 경우를 제외하고는 농도 의존적으로 COX-2의 발현이 증가하였, AP-1 역시 농도가 증가함에 따라 그 발현양이 증가하였다. 반면, 실시예 1(b)에서는 0.02% 처리한 경우를 제외하고는 농도 의존적으로 COX-2의 발현이 감소하였고, AP-1도 농도가 증가함에 따라 확연히 감소하였다.As shown in Figure 5 and 6, in Comparative Example (a) except for 0.02% treatment, the expression of COX-2 increased in a concentration-dependent manner, AP-1 also increased the amount of expression as the concentration increases It was. On the other hand, in Example 1 (b) except for 0.02% treatment, the expression of COX-2 decreased in a concentration-dependent manner, AP-1 also decreased significantly as the concentration increased.
이를 종합하면, 실시예 1은 피부 염증 반응 인자를 감소시켜 피부 염증을 억제할 수 있다는 것을 알 수 있다.
Taken together, it can be seen that Example 1 can suppress skin inflammation by reducing skin inflammatory response factors.
3. 피부염 동물 모델에 대한 피부 염증 증상 개선 효능 평가3. Evaluation of Skin Inflammation Symptom Improvement Efficacy in Dermatitis Animal Model
아토피 피부염을 유발시킨 쥐에 대하여 음용수에 추출물을 첨가하지 않은 대조군(control), 비교예를 첨가한 비교군 및 실시예 1을 첨가한 실험군으로 나누어 염증 증상의 개선 정도를 평가하고, 그 결과를 도 7 내지 도 11에 나타냈다. 각 사진에서 A는 대조군, B는 비교군, C는 실험군의 동물 사진이다.The rats causing atopic dermatitis were divided into a control group without adding an extract to drinking water, a comparative group with a comparative example, and an experimental group with Example 1 to evaluate the degree of improvement of the inflammatory symptoms. 7 to 11 are shown. In each photo, A is a control group, B is a comparison group, and C is an animal photograph of the experimental group.
상기 실험 방법에 따라 NC/Nga mice에 DNCB를 처리하였을 때 4일 후부터 처리 부위에 피부 발진이 확연히 나타나기 시작했다(도 7 참조). 투여 1주 후에는, 대조군(A)은 피부 발진이 심해졌으며 비교군(B)과 실험군(C) 역시 대조군(A) 보다는 약하지만 피부 발진이 진행됨을 관찰할 수 있었다(도 8 참조). 투여 2주 후에는, 대조군(A)의 피부 발진은 더욱 악화 되었으나, 비교군(B)과 실험군(C) 모두에서 피부 발진이 조금씩 개선되는 증상을 관찰할 수 있었고, 특히 실험군(C)에서의 개선 정도가 다소 높음을 확인할 수 있었다(도 9 참조). 투여 3주 후에는, 대조군(A)의 피부 발진이 더 이상 악화되지는 않았으나, 비교군(B)과 실험군(C)에서 피부 발진이 상당히 많이 회복됨을 볼 수 있었다(도 10 참조). 투여 4주 후에는, 대조군(A)의 피부 발진도 다소 개선됨을 볼 수 있었으며 비교군(B)과 실험군(C)에서 피부발진이 거의 정상 수준으로 회복되었음을 관찰할 수 있었다(도 11 참조).According to the experimental method, when the DNCB was treated in NC / Nga mice, the skin rash began to appear clearly at 4 days after treatment (see FIG. 7). After 1 week of administration, the control group (A) had a severe skin rash, and the control group (B) and the experimental group (C) were also weaker than the control group (A), but it was observed that the skin rash progressed (see FIG. 8). After 2 weeks of administration, the skin rash in the control group (A) became worse, but the symptoms of the skin rash improved slightly in both the control group (B) and the experimental group (C), especially in the experimental group (C). It was confirmed that the degree of improvement is rather high (see FIG. 9). After 3 weeks of administration, the skin rash of the control group (A) did not worsen anymore, but it was seen that the skin rash recovered significantly in the comparison group (B) and the experimental group (C) (see FIG. 10). After 4 weeks of administration, the skin rash of the control group (A) was also slightly improved, and it was observed that the skin rash was restored to almost normal level in the comparison group (B) and the experimental group (C) (see FIG. 11).
상기 실험 결과를 점수화하여 평가한 결과(Clinical skin severity score)를 도 12에 나타냈다. 도 12에 나타나 있는 것처럼, 투여 2주후 대조군(A)(8.5) 보다 비교군(B)(3.8) 및 실험군(C)(3.5)의 피부 손상이 더욱 감소하였다. 투여 3주차에는 대조군(A)의 피부손상도는 9.5로 투여 2주차 때보다 다소 심해진 반면, 비교군(B)(3.1)과 실험군(C)(2.8)에서는 모두 피부 발진 및 손상 정도가 지속적으로 회복되어 갔다. 마지막 주인 투여 4주차에는, 대조군(A)에서도 피부 손상도가 6.5 정도로 되어 투여 3주차 때보다 많은 회복을 보였으나, 비교군(B)(1.5)과 실험군(C)(1.2)은 거의 정상으로 개선되었다. 또한 전체 실험 과정을 통해 실험군(C)에서 피부 염증 완화 및 회복이 초기부터 빠르게 나타난 것을 알 수 있고, 회복 정도 또한 비교군(B)에 비하여 실험군(C)이 큰 것을 알 수 있다.
The results obtained by scoring the test results (Clinical skin severity score) are shown in FIG. 12. As shown in FIG. 12, two weeks after administration, the skin damage of the control group (B) (3.8) and the experimental group (C) (3.5) was further reduced than the control group (A) (8.5). At 3 weeks of administration, the skin damage of the control group (A) was 9.5, which was slightly worse than that of the 2nd week of administration. I recovered. At
4. 피부염 동물 모델에서 추출물이 4. Extracts from dermatitis animal models 염증인자Inflammatory factor 및 And 면역인자Immune factor 발현에 미치는 영향 Effect on expression
아토피 피부염을 유발시킨 쥐에 대하여 음용수에 추출물을 첨가하지 않은 대조군(control), 비교예를 첨가한 비교군 및 실시예 1을 첨가한 실험군으로 나누어 혈청 내 여러 염증인자 및 면역인자 발현에 미치는 영향을 평가하였다.
The rats inducing atopic dermatitis were divided into a control group without adding extract to drinking water, a comparative group with a comparative example, and an experimental group with Example 1, and their effects on the expression of various inflammatory and immune factors in serum. Evaluated.
1) 면역글로불린E(IgE)에 대한 결과1) Results for Immunoglobulin E (IgE)
면역글로불린E(IgE)에 대한 결과는 도 13에 나타내었다. 실험 초기에는 아토피 피부염 유발로 인하여 대조군, 비교군 및 실험군 모두에서 면역글로불린의 수치가 서로 유사한 정도로 높은 것을 알 수 있다. 대조군(control)의 혈청 중 IgE 생성량은 투여 3주 동안 지속적으로 증가하였으나 4주차에는 약간 감소하였다. 비교군에서 혈청 중 IgE 생성량은 1주차에 피부염 유발로 인해 급격히 증가한 후 투여시간경과에 따라 크게 감소되었다. 실험군의 IgE 생성량은 1주차에 피부염 유발로 인해 급격히 증가한 후 투여시간경과에 따라 통계적으로 유의성 있게 크게 감소되었다(p<0.01). 특히 실험군은 비교군과 동일한 경향을 보이기는 하였으나, 실험군의 혈청 중 IgE 생성량 감소폭은 더 컸다. 각 실험 기간별 면역글로불린 IgE의 구체적인 수치는 하기 표와 같다.
Results for immunoglobulin E (IgE) are shown in FIG. 13. Early in the experiment, due to the induction of atopic dermatitis, it can be seen that the levels of immunoglobulins are similar to each other in the control, comparison and experimental groups. IgE production in control serum was continuously increased during 3 weeks of administration but slightly decreased at 4 weeks. Serum IgE production increased rapidly due to dermatitis in the first week and then decreased significantly with time of administration. The IgE production in the experimental group increased sharply due to dermatitis in the first week and then significantly decreased significantly with the administration time (p <0.01). In particular, the experimental group showed the same tendency as the comparative group, but the decrease in IgE production in the serum of the experimental group was greater. Specific values of immunoglobulin IgE for each experimental period are shown in the following table.
2) 인터루킨-4(IL-4)에 대한 결과2) Results for Interleukin-4 (IL-4)
인터루킨-4(IL-4)에 대한 결과는 도 14에 나타내었다. 도 14에 나타나 있는 것처럼, 대조군은 45.25±4.85 pg/㎖, 비교군은 25.12±2.9 pg/㎖, 실험군은 20.35±3.54 pg/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 감소를 나타냈으며, 그 중에서도 실험군에서의 IL-4 감소폭이 더 컸다.
Results for interleukin-4 (IL-4) are shown in FIG. 14. As shown in FIG. 14, the control group was 45.25 ± 4.85 pg / ml, the comparison group was 25.12 ± 2.9 pg / ml, and the experimental group was 20.35 ± 3.54 pg / ml. Both the control and experimental groups showed a significant decrease (p <0.01) compared to the control group, and the reduction of IL-4 in the experimental group was greater.
3) 인터루킨-5(IL-5)에 대한 결과3) Results for Interleukin-5 (IL-5)
인터루킨-5(IL-5)에 대한 결과는 도 15에 나타내었다. 도 15에 나타나 있는 것처럼, 대조군은 215.63±5.22 pg/㎖, 비교군은 156.52±4.12 pg/㎖, 실험군은 145.66±4.52 pg/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 감소를 나타냈으며, 그 중에서도 실험군에서의 IL-5 감소폭이 더 컸다.
Results for interleukin-5 (IL-5) are shown in FIG. 15. As shown in FIG. 15, the control group was 215.63 ± 5.22 pg / ml, the comparative group was 156.52 ± 4.12 pg / ml, and the experimental group was 145.66 ± 4.52 pg / ml. Both the control and experimental groups showed a significant decrease (p <0.01) compared to the control group, and the reduction of IL-5 in the experimental group was greater.
4) 인터루킨-6(IL-6)에 대한 결과4) Results for Interleukin-6 (IL-6)
인터루킨-6(IL-6)에 대한 결과는 도 16에 나타내었다. 도 16에 나타나 있는 것처럼, 대조군은 450.12±4.25 pg/㎖, 비교군은 256.45±4.75 pg/㎖, 실험군은 241.54±5.23 pg/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 감소를 나타냈으며, 그 중에서도 실험군에서의 IL-6 감소폭이 더 컸다.
Results for interleukin-6 (IL-6) are shown in FIG. 16. As shown in FIG. 16, the control group was 450.12 ± 4.25 pg / ml, the control group was 256.45 ± 4.75 pg / ml, and the experimental group was 241.54 ± 5.23 pg / ml. Both the control and experimental groups showed a significant decrease (p <0.01) compared to the control group, and the reduction of IL-6 was greater in the experimental group.
5) 면역글로불린M(IgM)에 대한 결과5) Results for Immunoglobulin M (IgM)
면역글로불린M(IgM)에 대한 결과는 도 17에 나타내었다. 도 17에 나타나 있는 것처럼, 대조군은 585.15±45.58 ㎍/㎖, 비교군은 325±19.21 ㎍/㎖, 실험군은 288±25.25 ㎍/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 감소를 나타냈으며, 그 중에서도 실험군에서의 IL-4 감소폭이 더 컸다.
Results for immunoglobulin M (IgM) are shown in FIG. 17. As shown in FIG. 17, the control group was 585.15 ± 45.58 μg / ml, the comparison group was 325 ± 19.21 μg / ml, and the experimental group was 288 ± 25.25 μg / ml. Both the control and experimental groups showed a significant decrease (p <0.01) compared to the control group, and the reduction of IL-4 in the experimental group was greater.
6) 면역글로불린G1(IgG1)에 대한 결과6) Results for Immunoglobulin G1 (IgG1)
면역글로불린G1(IgG1)에 대한 결과는 도 18에 나타내었다. 도 18에 나타나 있는 것처럼, 대조군은 2,784±255.85 ㎍/㎖, 비교군은 1,958±125.25 ㎍/㎖, 실험군은 1,754±165.73 ㎍/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 감소를 나타냈으며, 그 중에서도 실험군에서의 IgG1 감소폭이 더 컸다.
Results for immunoglobulin G1 (IgG1) are shown in FIG. 18. As shown in FIG. 18, the control group was 2,784 ± 255.85 μg / ml, the comparison group was 1,958 ± 125.25 μg / ml, and the experimental group was 1,754 ± 165.73 μg / ml. Both control and experimental groups showed a significant decrease (p <0.01) compared to the control group, and the decrease of IgG1 in the experimental group was greater.
7) 인터페론감마(IFN-γ)에 대한 결과7) Results for Interferon-gamma (IFN-γ)
인터페론감마(IFN-γ)에 대한 결과는 도 19에 나타내었다. 도 19에 나타나 있는 것처럼, 대조군은 815.15±58.52 pg/㎖, 비교군은 1,158±85.65 pg/㎖, 실험군은 1,298±128.54 pg/㎖로 나타났다. 비교군과 실험군 모두 대조군에 비하여 유의성 있는(p<0.01) 증가를 나타냈으며, 그 중에서도 실험군에서의 IFN-γ 증가폭이 더 컸다. 인터페론감마는 특히 항암물질로 널리 알려져 있는 물질이다.
Results for interferon gamma (IFN-γ) are shown in FIG. 19. As shown in FIG. 19, the control group was 815.15 ± 58.52 pg / ml, the comparison group was 1,158 ± 85.65 pg / ml, and the experimental group was 1,298 ± 128.54 pg / ml. Both the control and experimental groups showed a significant increase (p <0.01) compared to the control group, and the IFN-γ increase in the experimental group was greater. Interferon gamma is particularly well known as an anticancer substance.
5. 5. 제형예의Formulation 제조 Produce
통상의 기능성 식품(건강기능식품) 제조방법에 따라 하기의 성분을 혼합하여 다양한 제형의 건강기능식품을 제조한다. 하기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강기능식품에 적합한 성분을 영양권장량에 준하여 제형예로 조성하였으나, 그 배합비를 임의로 변형하여 실시하여도 무방하다.
According to the conventional functional food (health functional food) manufacturing method by mixing the following ingredients to produce a health functional food of various formulations. The composition ratio of the following vitamin and mineral mixtures was formulated as an example of the formulation based on the nutritional recommended amount of a relatively suitable component for health functional food, but may be carried out by modifying the formulation ratio arbitrarily.
<제형예 1> 음료<Formulation Example 1> Drink
하기 분량의 성분을 혼합액을 2∼3㎛의 필터를 통과시켜 청징화한 최종혼합액을 90∼93℃에서 15∼20초간 전 살균하여 50㎖병에 충진하고 80∼85℃에서 15∼20분간 후 살균하여 음료제품(50㎖)을 제조한다.The following amount of the component was passed through a filter having a diameter of 2-3 μm, and the final mixture was clarified at 90-93 ° C. for 15-20 seconds to be sterilized in a 50 ml bottle, followed by 15-20 minutes at 80-85 ° C. Sterilization to prepare a beverage product (50ml).
음료1
Drinks1
히아루론산
콜라겐 펩티드
비타민 C
비타민 B2
비타민 B6
식이섬유
석류추출물
액상과당
정제수Example 1 Extract
Hyaluronic acid
Collagen Peptides
Vitamin C
Vitamin B 2
Vitamin B 6
Dietary Fiber
Pomegranate Extract
Liquid fructose
Purified water
75
75
75
4
3
1,000
8,000
20,000
적량1,000
75
75
75
4
3
1,000
8,000
20,000
Suitable amount
음료2
Drinks2
구연산
올리고당
타우린
정제수Example 1 Extract
Citric acid
oligosaccharide
Taurine
1000㎎
100g
1g
적량1000 mg
1000mg
100 g
1g
Suitable amount
<제형예 2> 정제<Formulation Example 2> Tablet
하기 분량의 재료를 혼합하고 반죽하여 과립제조기를 통과시켜 과립을 제조한 다음 스테아린산 마그네슘 5mg를 배합하여 고르게 혼합한 뒤 일정한 중량으로 타정하여 정제제품을 제조한다.The following amounts of ingredients are mixed and kneaded to produce granules through a granulation machine, and then 5 mg of magnesium stearate is mixed and evenly mixed and compressed into tablets to produce a tablet.
히아루론산
콜라겐 펩티드
비타민 C
비타민 B2
비타민 B6
식이섬유
유당
락츄로스
향료Example 1 Extract
Hyaluronic acid
Collagen Peptides
Vitamin C
Vitamin B2
Vitamin B6
Dietary Fiber
Lactose
Lacchuros
Spices
75
75
75
4
3
100
200
260
301,000
75
75
75
4
3
100
200
260
30
<제형예 3> 연질캅셀<Formulation Example 3> soft capsule
하기 분량의 재료를 혼합하고 균질화한 다음 통상의 방법에 따라 일정중량의 연질캅셀에 충전하여 제조한다.The following amount of material is mixed, homogenized and prepared by filling into a soft capsule of constant weight according to a conventional method.
히아루론산
콜라겐 펩티드
비타민 C
비타민 B2
비타민 B6
토코페롤
식이섬유
소맥배아유
밀납
왁스Example 1 Extract
Hyaluronic acid
Collagen Peptides
Vitamin C
Vitamin B2
Vitamin B6
Tocopherol
Dietary Fiber
Wheat germ oil
Wax
Wax
75
75
75
4
3
50
100
3.500
500
5001,000
75
75
75
4
3
50
100
3.500
500
500
<제형예 4> 과립제<Formulation Example 4> Granules
하기 분량의 재료를 혼합하고 반죽하여 과립제조기를 통과시켜 과립을 제조한다.The following amounts of materials are mixed and kneaded to produce granules through a granulator.
히아루론산
콜라겐 펩티드
비타민 C
비타민 B2
비타민 B6
식이섬유
유당
락츄로스
향료Example 1 Extract
Hyaluronic acid
Collagen Peptides
Vitamin C
Vitamin B2
Vitamin B6
Dietary Fiber
Lactose
Lacchuros
Spices
75
75
75
4
3
100
200
260
301,000
75
75
75
4
3
100
200
260
30
<제형예 5> 분말제<Formulation Example 5> Powder
하기 분량의 재료를 혼합하여 통상의 제조방법을 사용하여 분말제를 제조한다.The powder of the following amount is mixed and a powder is manufactured using a conventional manufacturing method.
비타민 A 아세테이트
비타민 E
비타민 B1
비타민 B2
비타민 B6
나이아신
엽산
황산제1철
산화아연
탄산마그네슘
인산칼슘
구연산칼륨
염화마그네슘Example 1 Extract
Vitamin A Acetate
Vitamin E
Vitamin B1
Vitamin B2
Vitamin B6
Niacin
Folic acid
Ferrous sulfate
Zinc oxide
Magnesium carbonate
Calcium phosphate
Potassium citrate
Magnesium chloride
70㎍
1㎎
0.2㎎
0.2㎎
0.2㎎
20㎎NE
50㎍
2㎎
1㎎
20㎎
100㎎
100㎎
25㎎1,000mg
70 µg
1mg
0.2mg
0.2mg
0.2mg
20mgNE
50 µg
2mg
1mg
20mg
100mg
100mg
25mg
<제형예 6> 시럽제<Formulation Example 6> Syrup
하기 분량의 재료를 혼합하여 통상의 제조방법을 사용하여 시럽제를 제조한다.The following amounts of materials are mixed to prepare syrups using conventional manufacturing methods.
전분 글리콜산나트륨
히드록시프로필메틸셀룰로오스
잔탄검
D-만니톨
정제백당
구연산
구연산나트륨Example 1 Extract
Starch sodium glycolate
Hydroxypropylmethylcellulose
Xanthan Gum
D-mannitol
Refined sugar
Citric acid
Sodium citrate
1.4㎎
3.2㎎
0.3㎎
50㎎
142㎎
0.3㎎
0.5㎎0.2mg
1.4mg
3.2mg
0.3mg
50 mg
142mg
0.3mg
0.5mg
<제형예 7> 껌<Formulation Example 7> Gum
하기 분량의 재료를 혼합하고 반죽하여 통상의 방법으로 껌을 제조한다.The following amounts of ingredients are mixed and kneaded to prepare a gum in a conventional manner.
껌베이스
설탕
향료
물Example 1 Extract
Gum base
Sugar
Spices
water
20
72
1
25
20
72
One
2
<제형예 8> 비스켓Formulation Example 8 Biscuits
하기 분량의 재료를 혼합하고 반죽하여 통상의 방법으로 비스켓을 제조한다.The following amounts of ingredients are mixed and kneaded to prepare biscuits in a conventional manner.
박력 1급
중력 1급
정백당
식염
포도당
팜쇼트닝
중조
중아황산나트륨
쌀가루
비타민 B1
비타민 B2
밀크향
물
전지분유
대용분유
제일인산칼슘
살포염
분무유Example 1 Extract
Gravity Class I
White sugar
saline
glucose
Palm Shortening
baking soda
Sodium bisulfite
Rice flour
Vitamin B1
Vitamin B2
Milk flavor
water
Whole milk powder
Substitute powdered milk
Calcium Phosphate
Spreading salt
Spray oil
21.59
22.22
4.80
0.73
0.78
11.15
0.17
0.16
1.45
0.0001
0.0001
0.04
21.33
1.16
0.29
0.03
0.29
7.275.00
21.59
22.22
4.80
0.73
0.78
11.15
0.17
0.16
1.45
0.0001
0.0001
0.04
21.33
1.16
0.29
0.03
0.29
7.27
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시 양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항 들과 그것들의 등가물에 의하여 정의된다고 할 것이다.While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.
Claims (10)
Health functional food composition for improving skin diseases, including extracts of mammundong, ginseng, Schisandra chinensis and Eoseongcho.
상기 추출물은 조추출액, 조추출액의 건조물, 조추출액의 농축물. 조추출액의 분획물, 분획물의 건조물 및 분획물의 농축물 중에서 선택된 하나 이상인 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물.
The method of claim 1,
The extract is crude extract, dried extract of crude extract, concentrate of crude extract. A dietary supplement for improving skin disease, characterized in that at least one selected from fractions of crude extract, dried fractions and concentrates of fractions.
상기 맥문동, 인삼, 오미자 및 어성초의 추출물은 맥문동 : 인삼 : 오미자 : 어성초의 중량비율이 1 내지 8 : 1 내지 4 : 1 내지 4 : 1 내지 8 인 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물.
The method of claim 1,
The extracts of the mammun-dong, ginseng, Schisandra chinensis and Eochocho are the weight ratio of McMoon-dong: Ginseng: Schisandra chinensis: Echo vinegar 1 to 8: 1 to 4: 1 to 4: 1 to 8, characterized in that the health functional food composition for improving skin diseases .
상기 맥문동, 인삼, 오미자 및 어성초의 추출물은 맥문동 : 인삼 : 오미자 : 어성초의 중량비율이 1 : 1 : 1 : 2 인 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물.
The method of claim 1,
The extracts of the mammundong, ginseng, Schisandra chinensis and Eoseongcho is the weight ratio of McMoondong: Ginseng: Schisandra chinensis: Echo vinegar is 1: 1: 1: 2, a health functional food composition for improving skin diseases.
상기 추출물은 물, 탄소수 1 내지 탄소수 4의 저급 알코올 또는 이들의 혼합 용매로부터 선택된 용매로 가용한 추출물인 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물.
The method of claim 1,
The extract is a health functional food composition for improving skin diseases, characterized in that the extract available in a solvent selected from water, lower alcohols having 1 to 4 carbon atoms or a mixed solvent thereof.
상기 피부 질환 개선용 건강기능식품 조성물은 담체, 희석제, 부형제 및 첨가제 중 하나 이상을 더 포함하여 정제, 환제, 산제, 과립제, 분말제, 캡슐제 및 액제 제형으로 이루어진 군에서 선택된 하나의 제형으로 제형된 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물.
The method according to any one of claims 1 to 5,
The health functional food composition for improving skin diseases is further formulated as one formulation selected from the group consisting of tablets, pills, powders, granules, powders, capsules and liquid formulations further comprising one or more of carriers, diluents, excipients and additives. Health functional food composition for improving the skin disease, characterized in that.
상기 추출물을 농축하는 단계를 포함하는 피부 질환 개선용 건강기능식품 조성물의 제조방법.
Extracting by adding a solvent to ganmundong, ginseng, Schisandra chinensis and Eoseongcho to obtain an extract; And
Method for producing a health functional food composition for improving skin diseases comprising the step of concentrating the extract.
상기 추출물을 수득하는 단계는 맥문동, 인삼, 오미자 및 어성초의 총 중량대비 5 내지 20배 중량의 용매를 가하여 추출되는 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물의 제조방법.
The method of claim 7, wherein
The step of obtaining the extract is a method for producing a health functional food composition for improving skin diseases, characterized in that the extract by adding a solvent of 5 to 20 times the weight of the total weight of the mammundong, ginseng, Schisandra chinensis and Eoseongcho.
상기 추출물을 수득하는 단계는 80℃ 내지 120℃에서 1시간 내지 10시간 동안 수행되는 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물의 제조방법.
The method of claim 7, wherein
The step of obtaining the extract is a method for producing a health functional food composition for improving skin diseases, characterized in that performed for 1 hour to 10 hours at 80 ℃ to 120 ℃.
상기 농축하는 단계는 상기 추출물을 여과하는 단계; 상기 여과된 추출물을 감압 농축하는 단계, 상기 농축된 추출물을 건조하는 단계; 상기 건조된 농축물을 분쇄하여 분말화 하는 단계를 포함하는 것을 특징으로 하는 피부 질환 개선용 건강기능식품 조성물의 제조방법.The method of claim 7, wherein
The step of concentrating includes filtering the extract; Concentrating the filtered extract under reduced pressure, and drying the concentrated extract; Method for producing a health functional food composition for improving skin diseases, characterized in that it comprises the step of pulverizing the dried concentrate.
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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KR20160091507A (en) | 2015-01-23 | 2016-08-03 | 부산대학교 산학협력단 | Composition for treating, improving or preventing allergy disease |
KR101907921B1 (en) * | 2017-12-22 | 2018-10-16 | 한국 한의학 연구원 | Composition for preventing or ameliorating skin wrinkle comprising Schisandra chinensis extract as effective component |
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KR20220089193A (en) | 2020-12-21 | 2022-06-28 | 농업회사법인 주식회사 아그로스 | Functional composition for skin moisturizing and skin soothing |
KR20230011764A (en) | 2021-07-14 | 2023-01-25 | 농업회사법인 주식회사 아그로스 | Functional cosmetic composition for improving skin condition containing oriental herbal extracts |
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KR101483440B1 (en) * | 2008-05-02 | 2015-01-19 | (주)아모레퍼시픽 | Medicinal plants extract using processing of herbal medicine and composition of skin external application comprising the same |
KR20110086894A (en) * | 2010-01-25 | 2011-08-02 | 김선일 | Cosmetic included to alkaline powder & solution of shell |
KR101805004B1 (en) * | 2010-04-06 | 2017-12-07 | (주)아모레퍼시픽 | Fermentation method using medicinal leaf and skin external composition containing fermented extract by the method |
KR101200317B1 (en) * | 2010-06-03 | 2012-11-12 | 강원도 | Manufacturing Method for Nutritional Snack Using Puffing Cereal and Snack Molding-Syrup |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
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KR20160091507A (en) | 2015-01-23 | 2016-08-03 | 부산대학교 산학협력단 | Composition for treating, improving or preventing allergy disease |
KR101907921B1 (en) * | 2017-12-22 | 2018-10-16 | 한국 한의학 연구원 | Composition for preventing or ameliorating skin wrinkle comprising Schisandra chinensis extract as effective component |
WO2019124862A1 (en) * | 2017-12-22 | 2019-06-27 | 한국 한의학 연구원 | Skin wrinkle prevention and improvement composition containing schisandra chinensis extract as active ingredient |
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