KR102669645B1 - Cosmetic Composition for stress relief and skin barrier improvement comprising extract of Momordica charantia - Google Patents
Cosmetic Composition for stress relief and skin barrier improvement comprising extract of Momordica charantia Download PDFInfo
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- KR102669645B1 KR102669645B1 KR1020230098473A KR20230098473A KR102669645B1 KR 102669645 B1 KR102669645 B1 KR 102669645B1 KR 1020230098473 A KR1020230098473 A KR 1020230098473A KR 20230098473 A KR20230098473 A KR 20230098473A KR 102669645 B1 KR102669645 B1 KR 102669645B1
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- extract
- bitter melon
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Botany (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Cosmetics (AREA)
Abstract
본 발명은 여주 추출물을 유효성분으로 함유하는 스트레스 완화 및 피부 장벽 개선용 화장료 조성물에 관한 것으로, 본 발명의 화장료 조성물은 스트레스 호르몬인 코르티솔 생성 및 코르티솔 생성 조절 효소인 11β-HSD1 발현을 억제하여 스트레스 완화 효과를 나타내고, 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin)의 발현 회복 효과를 발휘함에 따라 스트레스에 의해 무너진 피부 장벽 개선 효과가 우수하다. The present invention relates to a cosmetic composition for relieving stress and improving the skin barrier containing bitter melon extract as an active ingredient. The cosmetic composition of the present invention relieves stress by suppressing the production of cortisol, a stress hormone, and the expression of 11β-HSD1, an enzyme that regulates cortisol production. It is effective in improving the skin barrier damaged by stress as it exerts a restoration effect on the expression of aquaporin-3 (AQP-3) and filaggrin.
Description
본 발명은 여주 추출물을 유효성분으로 함유하는 스트레스 완화 및 피부 장벽 개선용 화장료 조성물에 관한 것이다. 본 발명의 화장료 조성물은 스트레스 호르몬인 코르티솔의 생성을 억제하고, 코르티솔 생성 조절 효소인 11β-HSD1의 발현을 저해하여 스트레스 완화 효과를 보이며, 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin)의 발현을 촉진하여 스트레스로 인해 손상된 피부 장벽 개선에 효과적이다.The present invention relates to a cosmetic composition for relieving stress and improving the skin barrier containing bitter melon extract as an active ingredient. The cosmetic composition of the present invention shows a stress relieving effect by inhibiting the production of cortisol, a stress hormone, and inhibiting the expression of 11β-HSD1, an enzyme that regulates cortisol production, and aquaporin-3 (AQP-3) and filaggrin. It is effective in improving the skin barrier damaged by stress by promoting the expression of .
스트레스는 신체적 혹은 심리적으로 상당한 정도의 부담을 받아서 생기는 개인의 긴장 상태 혹은 반응을 말하며 스트레스 반응은 주관적 감정으로 개인의 차이가 크기 때문에 예방법이 정확히 구분되지 않는다. 그러나 스트레스를 적절히 예방하여 대응하지 못할 경우 우울증이나 불안 같은 심리적 스트레스를 유발할 뿐만 아니라 호르몬 변화, 심혈관 질환, 염증 및 면역기능 저하 같은 생리적 변화를 유발할 수 있다. 스트레스는 중추신경계, 자율신경계 및 시상하부-뇌하수체-부신축(HPA axis)을 활성화시켜 글루코코르티코이드(glucocorticoid), 부신피질자극호르몬(adrenocorticotropic hormone, ACTH), 부신피질호르몬방출인자(corticotropic releasing factor, CRF)의 분비를 증가시킨다. 또한, HPA axis의 활성화는 면역세포의 활동을 억제하고 림프구의 감소를 유발하여 면역기능을 저하시킨다. 그 중, 글루코코르티코이드는 면역, 성장, 발달, 대사 및 체내 항상성에 중요한 역할을 하고 콜레스테롤로부터 합성되는 스테로이드 호르몬으로 코르티솔(cortisol), 덱사메타손(dexamethasone), 프레드니손(prednisone), 프레드니솔론(prednisolone), 알도스테론(aldosterone) 등이 있다. 코르티솔은 스트레스 반응에서 가장 먼저 반응하는 호르몬으로 대표적인 스트레스 호르몬으로 알려져 있다. Stress refers to an individual's state of tension or reaction that occurs due to a significant amount of physical or psychological strain, and because the stress response is a subjective emotion and varies greatly from person to person, prevention methods are not clearly defined. However, if stress is not properly prevented and responded to, it can not only cause psychological stress such as depression or anxiety, but also cause physiological changes such as hormonal changes, cardiovascular disease, inflammation, and decreased immune function. Stress activates the central nervous system, autonomic nervous system, and hypothalamic-pituitary-adrenal axis (HPA axis), releasing glucocorticoids, adrenocorticotropic hormone (ACTH), and corticotropic releasing factor (CRF). ) increases the secretion of Additionally, activation of the HPA axis suppresses the activity of immune cells and causes a decrease in lymphocytes, thereby lowering immune function. Among them, glucocorticoids play an important role in immunity, growth, development, metabolism, and body homeostasis. They are steroid hormones synthesized from cholesterol and include cortisol, dexamethasone, prednisone, prednisolone, and aldosterone ( aldosterone), etc. Cortisol is the hormone that reacts first in the stress response and is known as a representative stress hormone.
코르티솔은 스트레스 반응에서 가장 먼저 반응하는 호르몬으로 대표적인 스트레스 호르몬으로 알려져 있다. 코르티솔은 HPA axis와 11β- HSD(hydroxysteroid dehydrogenase)라는 효소 활성에 의해 생성이 조절된다. 그 중, 11β- HSD는 NADPH 의존성 효소로 2종류의 동위효소가 존재한다. 11β- HSD 1은 간, 지방, 근육 등에서 높게 발현되고 체내에서 비활성의 상태인 코르티손(cotisone)을 활성형인 코르티솔로 전환시킨다. 11β- HSD 2는 신장, 간, 폐 등에서 발현되고 코르티솔을 코르티손으로 산화시키는 역할을 한다. 이 2종류의 효소 작용에 의해 체내에서 코르티솔의 항상성이 조절되며 코르티솔의 수준을 확인하여 스트레스 정도를 확인하는 지표로 사용된다. 또한, 코르티솔은 피부에 존재하는 각질형성세포, 섬유아세포, 멜라닌형성세포 등과 같은 다양한 세포에서 생성되며 코르티솔의 증가는 면역력을 약화시키고 피부 세포의 밀착연접(tight junction)을 억제하여 피부장벽의 기능을 손상시키는 것이 보고되었다. 최근 연구에서 스트레스를 받는 상황에서 정상 상태일 때보다 코르티솔 함량 및 11β- HSD 1의 발현이 증가함이 확인되었다. 또한, 피부장벽 단백질의 감소로 인해 경피수분손실이 증가하였고 이를 통해 코르티솔이 피부 장벽에 영향을 주는 것을 입증하였다.Cortisol is the hormone that reacts first in the stress response and is known as a representative stress hormone. The production of cortisol is regulated by the HPA axis and the activity of an enzyme called 11β-HSD (hydroxysteroid dehydrogenase). Among them, 11β-HSD is an NADPH-dependent enzyme and two types of isoenzymes exist. 11β-HSD 1 is highly expressed in the liver, fat, and muscles and converts inactive cortisone into active cortisol in the body. 11β-HSD 2 is expressed in the kidneys, liver, lungs, etc. and plays a role in oxidizing cortisol to cortisone. The action of these two types of enzymes regulates cortisol homeostasis in the body and is used as an indicator to check the level of stress by checking the level of cortisol. In addition, cortisol is produced in various cells such as keratinocytes, fibroblasts, and melanocytes present in the skin, and an increase in cortisol weakens immunity and inhibits the tight junctions of skin cells, thereby impairing the function of the skin barrier. Damage has been reported. In a recent study, it was confirmed that cortisol content and expression of 11β-HSD 1 increased in stressful situations compared to normal conditions. In addition, transepidermal water loss increased due to a decrease in skin barrier proteins, proving that cortisol affects the skin barrier.
현대인들은 다양한 심리적, 정신적 및 신체적 스트레스 상황에 놓여 있다. 스트레스로 인하여 면역력 저하 및 다양한 피부 기능 저하 현상이 나타남에 따라 스트레스를 완화 시킬 수 있는 방법에 대한 관심이 높아지고 있다. 현재 스트레스 완화를 위해 향기를 이용한 아로마 테라피, 마사지 등의 신체적, 정신적 대체 요법이 사용되고 있으나 스트레스로 인한 피부 손상을 완화할 수 있는 제품 등의 개발은 미비한 실정으로 피부에 안전한 천연물을 활용하여 피부 손상을 개선시킬 수 있는 소재의 개발이 필요하다.Modern people are subject to various psychological, mental and physical stress situations. As stress causes decreased immunity and various skin functions, interest in ways to relieve stress is increasing. Currently, physical and mental alternative therapies such as aromatherapy and massage using scents are being used to relieve stress, but the development of products that can alleviate skin damage caused by stress is insufficient, so natural products that are safe for the skin are being used to prevent skin damage. Development of materials that can be improved is necessary.
본 발명은 여주 추출물을 유효성분으로 함유하여 스트레스 완화 및 스트레스로 인해 손상된 피부 장벽 개선 효과를 나타내는 화장료 조성물을 제공하고자 한다. The present invention seeks to provide a cosmetic composition that contains bitter melon extract as an active ingredient and has the effect of relieving stress and improving the skin barrier damaged by stress.
본 발명은 여주(Momordica charantia) 에탄올 추출물을 유효성분으로 함유하는 피부 장벽 손상 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin barrier damage containing ethanol extract of Momordica charantia as an active ingredient.
본 발명은 여주(Momordica charantia) 에탄올 추출물을 유효성분으로 함유하는 피부 스트레스 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin stress containing ethanol extract of Momordica charantia as an active ingredient.
본 발명에 있어서, 상기 추출물은, 여주의 꽃, 잎, 열매, 줄기 또는 뿌리 중 선택되는 어느 하나 이상의 부위에서 얻은 추출물인 것을 특징으로 하는 것일 수 있다.In the present invention, the extract may be an extract obtained from one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon.
본 발명에 있어서, 상기 추출물은, 용매 추출법, 초임계 추출법 및 초음파 추출법 중 선택되는 어느 하나의 추출법을 통해 추출하는 것을 특징으로 하는 것일 수 있다.In the present invention, the extract may be characterized in that it is extracted through any one extraction method selected from solvent extraction, supercritical extraction, and ultrasonic extraction.
본 발명은 코르티솔 생성 또는 11β-HSD1의 발현 억제 효과, AQP-3 또는 filaggrin의 발현 촉진 효과를 발휘하는 여주 추출물을 함유하여 스트레스 완화 및 스트레스로 인해 무너진 피부 장벽 개선에 효과적인 화장료 조성물을 제공한다. The present invention provides a cosmetic composition that is effective in relieving stress and improving the skin barrier collapsed due to stress by containing a bitter melon extract that exerts the effect of suppressing cortisol production or expression of 11β-HSD1 and promoting the expression of AQP-3 or filaggrin.
도 1은 여주 추출물의 인간유래 각질형성세포에서의 세포 독성을 확인한 결과이다.
도 2는 여주 추출물의 코르티솔 생성 억제 효과를 확인한 결과이다.
도 3은 여주 추출물의 11β-HSD1 발현 억제 효과를 확인한 결과이다.
도 4는 여주 추출물의 아쿠아포린-3(AQP-3) 발현 회복 효과를 확인한 결과이다.
도 5는 여주 추출물의 필라그린(filaggrin) 발현 회복 효과를 확인한 결과이다.Figure 1 shows the results of confirming the cytotoxicity of bitter melon extract on human-derived keratinocytes.
Figure 2 shows the results of confirming the inhibitory effect of bitter melon extract on cortisol production.
Figure 3 shows the results confirming the inhibitory effect of bitter melon extract on 11β-HSD1 expression.
Figure 4 shows the results confirming the effect of the bitter melon extract on restoring aquaporin-3 (AQP-3) expression.
Figure 5 shows the results confirming the effect of bitter melon extract on restoring filaggrin expression.
본 발명은 여주(Momordica charantia) 에탄올 추출물을 유효성분으로 함유하는 피부 장벽 손상 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin barrier damage containing ethanol extract of Momordica charantia as an active ingredient.
본 발명은 여주(Momordica charantia) 에탄올 추출물을 유효성분으로 함유하는 피부 스트레스 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin stress containing ethanol extract of Momordica charantia as an active ingredient.
여주(Momordica charantia)는 아시아, 아프리카 등 열대 지역에서 서식하는 덩굴성 한해살이 작물로 주로 관상용으로 이용해 왔다. 여주는 쓴맛을 가지고 있어 고과라고도 불리며 식용 및 약용으로 이용된다. 여주는 비타민 C, 비타민 A, 아미노산, 갈락트론산, 펙틴, 사포닌, β-카로틴 등 여러 가지 생리 활성 성분이 함유되어 있어 항산화 및 혈당 조절 효과가 높은 것으로 알려져 있다. 특히 과실과 종자에 함유되어 있는 카란틴은 인슐린 분비를 촉진하여 혈당을 강화시키는 효과가 있다고 보고되어 있다. 그 외에도 바이러스 및 곰팡이에 대한 항균 활성, 항염 효과 등이 알려져 있다. Bitter melon ( Momordica charantia ) is an annual vine crop that grows in tropical regions such as Asia and Africa and has been mainly used for ornamental purposes. Bitter melon has a bitter taste, so it is also called bitter gourd and is used for edible and medicinal purposes. Bitter melon is known to have high antioxidant and blood sugar control effects as it contains various bioactive ingredients such as vitamin C, vitamin A, amino acids, galactronic acid, pectin, saponin, and β-carotene. In particular, charantin contained in fruits and seeds has been reported to have the effect of strengthening blood sugar levels by promoting insulin secretion. In addition, it is known to have antibacterial and anti-inflammatory effects against viruses and fungi.
한편, 스트레스는 우울증, 수면장애, 불안 등과 같은 심리적 영향 뿐만 아니라 면역력 저하, 피부 염증, 산화적 스트레스 및 장벽 기능의 손상을 야기할 수 있다. 코르티솔은 스트레스 상황에서 가장 먼저 생성되는 호르몬으로 11β-HSD 효소 활성에 의해 생성이 조절된다. 스트레스로 인해 코르티솔이 증가됨에 따라 세포 연접에 관여하는 인자를 감소시켜 피부 장벽을 약화시키는 것으로 알려져 있다. Meanwhile, stress can cause not only psychological effects such as depression, sleep disorders, and anxiety, but also decreased immunity, skin inflammation, oxidative stress, and damage to barrier function. Cortisol is the first hormone produced in stressful situations, and its production is regulated by 11β-HSD enzyme activity. It is known that as cortisol increases due to stress, it weakens the skin barrier by reducing factors involved in cell junctions.
한편, 본 발명에 있어서, 상기 피부 장벽 손상은, 스트레스로 인한 코르티솔 생성 증가로 말미암은 것일 수 있다.Meanwhile, in the present invention, the skin barrier damage may be due to increased cortisol production due to stress.
한편, 본 발명의 여주 에탄올 추출물은 코르티솔 생성 억제 또는 코르티솔 생성 조절 효소인 11β-HSD1(hydroxysteroid dehydrogenase 1) 발현 억제 효과를 발휘하는 것일 수 있다. 또한, 본 발명의 여주 에탄올 추출물은 스트레스로 인해 감소된 피부 장벽 강화 및 피부 보습 관련 인자인 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin) 발현 회복 효과를 발휘하는 것일 수 있다. 본 발명의 실험예에서 본 발명의 여주 에탄올 추출물을 이용하여 실험한 결과, 스트레스 호르몬인 코르티솔의 생성 억제 효과, 코르티솔 생성 조절 효소인 11β-HSD1의 발현 억제 효과가 확인되었다. 또한, 스트레스로 인해 감소된, 피부 장벽 강화 및 피부 보습 관련 인자인 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin)의 발현 회복 효과가 확인되었다. 이를 통해, 본 발명의 여주 에탄올 추출물은 스트레스 완화 및 스트레스로 인해 무너진 피부 장벽 개선에 탁월한 효과를 발휘하는 점을 알 수 있었다.Meanwhile, the ethanol extract of bitter melon of the present invention may exert an effect of suppressing cortisol production or suppressing the expression of 11β-HSD1 (hydroxysteroid dehydrogenase 1), an enzyme that regulates cortisol production. In addition, the ethanol extract of bitter melon of the present invention may exert the effect of strengthening the skin barrier, which is reduced due to stress, and restoring the expression of aquaporin-3 (AQP-3) and filaggrin, which are factors related to skin moisturization. As a result of an experiment using the ethanol extract of bitter melon of the present invention in an experimental example of the present invention, the effect of suppressing the production of cortisol, a stress hormone, and the effect of suppressing the expression of 11β-HSD1, an enzyme that regulates cortisol production, were confirmed. In addition, the effect of restoring the expression of aquaporin-3 (AQP-3) and filaggrin, factors related to skin barrier strengthening and skin moisturization, which were reduced due to stress, was confirmed. Through this, it was found that the ethanol extract of bitter melon of the present invention has an excellent effect in relieving stress and improving the skin barrier broken down by stress.
따라서, 본 발명의 화장료 조성물은, 코르티솔의 생성 억제, 11β-HSD1의 발현 억제, 아쿠아포린-3(AQP-3)의 발현 증가 또는 필라그린(filaggrin)의 발현 증가 중 선택되는 어느 하나의 효능을 가지는 것일 수 있다.Therefore, the cosmetic composition of the present invention has any one effect selected from inhibiting the production of cortisol, inhibiting the expression of 11β-HSD1, increasing the expression of aquaporin-3 (AQP-3), or increasing the expression of filaggrin. It can be something to have.
본 발명에 있어서, 상기 여주 에탄올 추출물은 용매 추출법, 초임계 추출법 또는 초음파 추출법 중 선택되는 어느 하나의 추출법을 이용하여 추출할 수 있다. 다만, 바람직하게는 60~80%(v/v) 에탄올을 추출용매로 이용하여 추출하며, 더욱 바람직하게는 70%(v/v) 에탄올을 추출용매로 이용하여 추출한다. In the present invention, the ethanol extract of Yeoju extract can be extracted using any one extraction method selected from solvent extraction, supercritical extraction, or ultrasonic extraction. However, extraction is preferably performed using 60-80% (v/v) ethanol as the extraction solvent, and more preferably, extraction is performed using 70% (v/v) ethanol as the extraction solvent.
한편, 본 발명에 있어서, 상기 여주 에탄올 추출물은, 여주의 꽃, 잎, 열매, 줄기 또는 뿌리 중 선택되는 어느 하나 이상의 부위에서 얻은 추출물인 것을 특징으로 하는 것일 수 있다.Meanwhile, in the present invention, the ethanol extract of bitter melon may be characterized as being an extract obtained from any one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon.
또한, 상기 여주 에탄올 추출물은, 바람직하게 화장료 조성물 전체 중량에 대하여 0.0001~30.0 중량%가 함유되는 것이 좋다. 더욱 바람직하게는 화장료 조성물 전체 중량에 대해서 0.01~10 중량% 함유되는 것이 좋다. 여주 추출물의 함량이 0.0001 중량% 미만인 경우에는 피부 스트레스 완화 또는 피부 장벽 개선 효과가 미미하고, 30.0 중량% 초과할 경우 함유량 증가에 따른 뚜렷한 효과 증가가 보이지 않을 수 있다.In addition, the ethanol extract of bitter melon is preferably contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. More preferably, it is contained in 0.01 to 10% by weight based on the total weight of the cosmetic composition. If the content of bitter melon extract is less than 0.0001% by weight, the effect of relieving skin stress or improving the skin barrier is minimal, and if it exceeds 30.0% by weight, a clear increase in effect may not be seen as the content increases.
한편, 본 발명의 화장료 조성물에 포함되는 성분은 유효성분으로서 본 발명의 여주 에탄올 추출물 이외에 화장료 조성물에 통상적으로 이용되는 성분들을 포함할 수 있으며, 예컨대 항산화제, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통상적인 보조제, 그리고 담체를 포함한다.Meanwhile, the ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the ethanol extract of Yeoju of the present invention as an active ingredient, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and Contains conventional auxiliaries such as flavorings, and carriers.
본 발명의 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 팩, 마사지크림 및 스프레이 등으로 제형화될 수 있으나, 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양 크림, 마사지크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 폼, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제조될 수 있다.The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing products. , may be formulated into oils, packs, massage creams, sprays, etc., but are not limited thereto. More specifically, it can be manufactured in the form of softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, or powder.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, the carrier ingredient may include animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌글리콜 또는 소르비탄의 지방산 에스테르가 있다.When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, and propylene. These include fatty acid esters of glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소 결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a suspension, the carrier component includes water, a liquid diluent such as ethanol or propylene glycol, and a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester. , microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant, etc. can be used.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the cosmetic composition is a spray, chlorofluorohydride may be additionally used. May contain propellants such as carbon, propane/butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 계면활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, Fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.
본 발명의 화장료 조성물이 비누, 계면활성제 함유 클렌징 제형 또는 계면활성제 비함유 클렌징 제형일 경우, 피부에 도포한 후 닦아내거나 떼거나 물로 씻어낼 수도 있다. 구체적인 예로서, 상기 비누는 액상비누, 가루비누, 고형비누 및 오일비누이며, 상기 계면활성제 함유 클렌징 제형은 클렌징 폼, 클렌징 워터, 클렌징 수건 및 클렌징 팩이며, 상기 계면활성제 비 함유 클렌징 제형은 클렌징크림, 클렌징 로션, 클렌징 워터 및 클렌징 겔이며, 이에 한정되는 것은 아니다.When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it can be applied to the skin and then wiped off, removed, or washed with water. As a specific example, the soap is liquid soap, powdered soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is cleansing cream. , cleansing lotion, cleansing water, and cleansing gel, but are not limited thereto.
이하, 본 발명의 구성을 하기 실시예 및 실험예를 통해 구체적으로 설명하고자 한다. 다만, 본 발명의 권리범위가 하기 실시예 및 실험예에만 한정되는 것은 아니고, 그와 등가의 기술적 사상의 변형까지를 포함한다.Hereinafter, the configuration of the present invention will be described in detail through the following examples and experimental examples. However, the scope of the present invention is not limited to the following examples and experimental examples, and includes modifications of the technical idea equivalent thereto.
[실시예 1: 여주 추출물 제조][Example 1: Preparation of bitter melon extract]
여주(Momordica charantia)를 음건하여 파쇄한 후, 20배 중량의 70%(v/v)에탄올을 첨가하여 24시간 동안 상온에서 추출하였다. 이후, 추출물을 여과하고, 감압농축 및 동결건조하여 여주 추출물을 제조하였다.After the bitter melon ( Momordica charantia ) was dried in the shade and crushed, 20 times the weight of 70% (v/v) ethanol was added and extracted at room temperature for 24 hours. Afterwards, the extract was filtered, concentrated under reduced pressure, and freeze-dried to prepare a bitter melon extract.
[실험예 1: 여주 추출물의 세포 독성 확인][Experimental Example 1: Confirmation of cytotoxicity of bitter melon extract]
본 실험에서는 여주 추출물의 세포 독성을 확인하기 위하여, 인간 유래 각질형성세포에서 여주 추출물의 영향을 확인하였다.In this experiment, in order to confirm the cytotoxicity of the bitter melon extract, the effect of the bitter melon extract on human-derived keratinocytes was confirmed.
인간 유래 각질형성세포를 96 웰 플레이트 (well plate)에 2 × 104 cells/well로 분주하고 37℃, 5% CO2 배양기에서 하루 동안 세포를 안정화시켰다. 이후, 상기 실시예 1에서 제조한 여주 추출물을 10, 30, 50 ㎍/㎖ 농도로 희석하여 24시간 동안 반응시켰다. 5 mg/㎖ 농도의 MTT 시약을 각 웰에 20 ㎕씩 첨가하고 배양기에서 2시간 반응시켰다. 상층액을 모두 제거하고 DMSO 100 ㎕를 각 웰에 추가하여 세포를 완전히 용해시킨 후 550 nm에서 흡광도를 측정하였다. 한편, 세포 생존율은 무처리(untreated) 세포 대비하여 백분율로 계산하여 아래 표 1에 나타내었다. Human-derived keratinocytes were distributed at 2 × 10 4 cells/well in a 96 well plate, and the cells were stabilized for one day in an incubator at 37°C and 5% CO 2 . Afterwards, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml and reacted for 24 hours. 20 μl of MTT reagent at a concentration of 5 mg/ml was added to each well and reacted in an incubator for 2 hours. All supernatants were removed, 100 ㎕ of DMSO was added to each well to completely lyse the cells, and the absorbance was measured at 550 nm. Meanwhile, cell survival rate was calculated as a percentage compared to untreated cells and is shown in Table 1 below.
(여주 추출물)Example 1 (㎍/㎖)
(Bitter melon extract)
실험 결과, 표 1 및 도 1에서 보듯이, 본 발명의 여주 추출물은 50 ㎍/㎖의 농도까지 세포 독성을 보이지 않았다. 한편, 도 1은 여주 추출물의 세포 독성 확인 결과를 나타낸 그래프이다. As a result of the experiment, as shown in Table 1 and Figure 1, the bitter melon extract of the present invention did not show cytotoxicity up to a concentration of 50 μg/ml. Meanwhile, Figure 1 is a graph showing the results of confirming the cytotoxicity of the bitter melon extract.
[실험예 2: 여주 추출물의 코르티솔 생성 억제 효과 확인][Experimental Example 2: Confirmation of the inhibitory effect of bitter melon extract on cortisol production]
본 실험에서는 여주 추출물의 코르티솔 생성에 대한 영향을 확인하고자 하였다.In this experiment, we sought to determine the effect of bitter melon extract on cortisol production.
인간 유래 각질형성세포를 24 웰 플레이트 (well plate)에 5 × 104 cells/well이 되도록 450 ㎕ 분주하고 37℃, 5% CO2 배양기에서 하루 동안 세포를 안정화시켰다. 코르티손(cortisone) 100 μM을 처리하여 스트레스 자극을 준 후, 상기 실시예 1에서 제조한 여주 추출물을 3, 10, 30, 50 ㎍/㎖ 농도로 희석하여 각각 처리한 후 24시간 동안 반응시켰다. 배지를 제거하고 1 × PBS로 3회 세척한 후 스크래퍼를 이용하여 세포를 수집한 뒤 초음파 처리하여 세포를 용해시켰다. 12,000 rpm, 4℃로 10분 동안 원심분리하여 상등액을 회수한 후 human cortisol elisa kit를 이용하여 세포내 코르티솔의 함량을 측정하였다. 아래 표 2는 코르티솔 생성 억제율을 백분율로 계산하여 나타낸 것이다.450 ㎕ of human-derived keratinocytes were dispensed into a 24-well plate at 5 × 10 4 cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO 2 for one day. After providing a stress stimulus by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 3, 10, 30, and 50 μg/ml, respectively, and reacted for 24 hours. The medium was removed, washed three times with 1×PBS, and the cells were collected using a scraper and sonicated to lyse the cells. The supernatant was recovered by centrifugation at 12,000 rpm and 4°C for 10 minutes, and the intracellular cortisol content was measured using a human cortisol elisa kit. Table 2 below shows the inhibition rate of cortisol production calculated as a percentage.
(여주 추출물)Example 1 (㎍/㎖)
(Bitter melon extract)
실험 결과, 표 2 및 도 2에서 보듯이, 본 발명의 여주 추출물은 농도 의존적으로 코르티솔의 생성을 억제하였다. 또한, 10 ㎍/㎖의 농도부터 28.18%로 유의한 억제율을 보였으며, 50 ㎍/㎖의 농도에서 최대 69.14%의 높은 코르티솔의 생성 억제 효과를 나타내는 점을 확인하였다 (도 2). 한편, 도 2는 여주 추출물의 코르티솔 생성 억제 효과 확인 결과를 나타낸 그래프이다.As a result of the experiment, as shown in Table 2 and Figure 2, the bitter melon extract of the present invention suppressed the production of cortisol in a concentration-dependent manner. In addition, a significant inhibition rate of 28.18% was observed at a concentration of 10 μg/ml, and a high cortisol production inhibition effect of up to 69.14% was confirmed at a concentration of 50 μg/ml (Figure 2). Meanwhile, Figure 2 is a graph showing the results of confirming the inhibitory effect of bitter melon extract on cortisol production.
[실험예 3 : 여주 추출물의 11β-HSD1 발현 억제 효과][Experimental Example 3: Inhibitory effect of 11β-HSD1 expression of bitter melon extract]
본 실험에서는 여주 추출물의 스트레스 완화 효과를 확인하기 위해 각질형성세포에서 코르티솔의 생성 조절 효소인 11β-HSD1에 대한 영향을 확인하였다. In this experiment, to confirm the stress-relieving effect of bitter melon extract, its effect on 11β-HSD1, an enzyme that regulates cortisol production in keratinocytes, was confirmed.
인간 유래 각질형성세포를 12 웰 플레이트 (well plate)에 3 × 105 cell/well로 분주하고 37℃, 5% CO2 배양기에서 하루 동안 세포를 안정화시켰다. 상기 실시예 1에서 제조한 여주 추출물을 10, 30, 50 ㎍/㎖ 농도로 희석하여 처리하였다. 6시간 동안 37℃, 5% CO2 배양기에서 반응시킨 후 단백질을 분리하였다. 상층액을 제거하고 1 × PBS로 한번 세척한 후 스크래퍼로 긁어 세포를 수집하였다. Cell lysis buffer를 이용하여 단백질을 분리하고 BCA assay로 단백질을 정량한 후 일정량의 단백질을 10% SDS-PAGE 겔에서 전기영동 시켰다. 겔 상의 분리된 단백질을 PVDF 멤브레인에 전이(transfer)시키고 5% 스킴밀크(skim milk)를 이용하여 1시간 동안 블로킹(blocking)하였다. 1 × TTBS 용액으로 3회 세척하고 1차 항체를 4℃에서 오버나잇(over night) 반응시킨 후, HRP가 결합된 2차 항체를 실온에서 2시간 동안 반응시켰다. 그 후, 1 × TTBS로 3 회 세척하고 ECL용액을 반응시켜 Chemi Doc 기기로 밴드를 검출하였다. Image J 1.47 software를 이용하여 수치화하였으며 베타-액틴(β-actin)의 발현을 기준으로 정량하였다. 한편, 아래 표 3은 무처리(Untreated) 세포 대비하여 11β-HSD1의 발현 억제율(%)을 계산하여 나타낸 것이다.Human-derived keratinocytes were distributed in a 12-well plate at 3 × 10 5 cells/well, and the cells were stabilized for one day in an incubator at 37°C and 5% CO 2 . The Yeoju extract prepared in Example 1 was diluted and treated at concentrations of 10, 30, and 50 μg/ml. After reacting in an incubator at 37°C and 5% CO 2 for 6 hours, proteins were separated. The supernatant was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. Proteins were separated using cell lysis buffer, proteins were quantified using BCA assay, and a certain amount of proteins were electrophoresed on a 10% SDS-PAGE gel. The separated proteins on the gel were transferred to a PVDF membrane and blocked for 1 hour using 5% skim milk. After washing three times with 1 Afterwards, it was washed three times with 1 × TTBS, reacted with ECL solution, and the band was detected with a Chemi Doc instrument. It was quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 3 below shows the calculated inhibition rate (%) of 11β-HSD1 expression compared to untreated cells.
(여주 추출물)Example 1 (㎍/㎖)
(Bitter melon extract)
실험 결과, 표 3 및 도 3에서 보듯이, 본 발명의 여주 추출물은 농도의존적으로 11β-HSD1의 발현을 억제하였다. 또한, 10 ㎍/㎖의 농도부터 52.47%로 유의적인 억제율을 보였으며 50 ㎍/㎖의 농도에서 최대 58.49%의 높은 11β-HSD1의 발현 억제 효과를 나타내는 점을 확인하였다. 한편, 도 3은 여주 추출물의 11β-HSD1 발현 억제 효과를 확인한 결과이다. As a result of the experiment, as shown in Table 3 and Figure 3, the bitter melon extract of the present invention suppressed the expression of 11β-HSD1 in a concentration-dependent manner. In addition, it was confirmed that a significant inhibition rate of 52.47% was observed at a concentration of 10 μg/ml, and that a high 11β-HSD1 expression inhibition effect of up to 58.49% was observed at a concentration of 50 μg/ml. Meanwhile, Figure 3 shows the results confirming the inhibitory effect of bitter melon extract on 11β-HSD1 expression.
[실험예 4 : 여주 추출물의 AQP-3 발현 회복 효과][Experimental Example 4: AQP-3 expression recovery effect of bitter melon extract]
본 실험에서는 여주 추출물의 스트레스에 인해 파괴된 피부 장벽 개선 효과를 확인하기 위해 피부 장벽 강화에 관여하는 인자인 아쿠아포린-3(AQP-3)에 대한 영향을 확인하였다. In this experiment, in order to confirm the effect of the bitter melon extract on improving the skin barrier damaged by stress, the effect on aquaporin-3 (AQP-3), a factor involved in strengthening the skin barrier, was confirmed.
각질형성세포를 6 웰 플레이트 (well plate)에 4 × 105 cell/well로 분주하고 37℃, 5% CO2 배양기에서 하루 동안 세포를 안정화시켰다. 코르티손(cortisone) 100 μM을 처리하여 스트레스 자극을 준 후, 상기 실시예 1에서 제조한 여주 추출물을 10, 30, 50 ㎍/㎖ 농도로 희석하여 각각 처리한 후 6시간 동안 반응시켰다. 배지를 제거하고 1 × PBS로 한번 세척한 후 스크래퍼로 긁어 세포를 수집하였다. 트리졸(Trizol)법을 이용하여 RNA를 분리하였다. Trizol 800 ㎕를 첨가하여 세포를 용해시키고 상온에서 5분 동안 반응시킨 후 클로로포름(chloroform) 200 ㎕를 첨가하여 볼텍싱(voltexing)한 다음 상온에서 5분 동안 반응시켰다. 12,000 rpm, 4℃로 15분 동안 원심분리하고 상등액만 새로운 튜브(tube)로 옮겨준 후 동량의 isopropanol을 첨가하여 1~3번 섞어주고 상온에서 10분 동안 반응시켰다. 12,000 rpm, 4℃로 15분 동안 원심분리하고 상등액을 모두 제거한 후 차가운 70% 에탄올을 800 ㎕씩 첨가하여 세척하고 다시 12,000 rpm, 4℃로 15분 동안 원심분리하였다. 상층액을 완전히 제거하고 펠릿(pellet)이 투명해질 때까지 건조시킨 후 RNA 양에 따라 DEPC DW를 첨가하였다. Nanodrop을 이용하여 RNA를 정량하여 PCR을 수행한 후 2% 아가로오스 겔 (agarose gel)에서 전기영동하여 밴드를 검출하였다. 밴드는 Image J 1.47 software를 이용하여 수치화하였으며 베타-액틴(β-actin)의 발현을 기준으로 정량하였다. 한편, 아래 표 4는 코르티손 처리군 대비하여 아쿠아포린-3(AQP-3)의 발현 회복율(%)을 계산하여 나타낸 것이다. Keratinocytes were distributed in a 6-well plate at 4 × 10 5 cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO 2 for one day. After providing stress stimulation by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml, respectively, and reacted for 6 hours. The medium was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. RNA was isolated using the Trizol method. Cells were lysed by adding 800 ㎕ of Trizol and reacted at room temperature for 5 minutes, then 200 ㎕ of chloroform was added, vortexed, and reacted at room temperature for 5 minutes. Centrifuge at 12,000 rpm, 4°C for 15 minutes, transfer only the supernatant to a new tube, add the same amount of isopropanol, mix 1 to 3 times, and react at room temperature for 10 minutes. After centrifugation at 12,000 rpm and 4°C for 15 minutes and removing all supernatants, 800 ㎕ of cold 70% ethanol was added to wash them, and centrifugation was performed again at 12,000 rpm and 4°C for 15 minutes. The supernatant was completely removed, the pellet was dried until it became transparent, and DEPC DW was added according to the amount of RNA. RNA was quantified using Nanodrop, PCR was performed, and the band was detected by electrophoresis on a 2% agarose gel. The bands were quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 4 below shows the calculated expression recovery rate (%) of aquaporin-3 (AQP-3) compared to the cortisone treatment group.
(여주 추출물)Example 1 (㎍/㎖)
(Bitter melon extract)
실험 결과, 표 4 및 도 4에서 보듯이, 코르티손 처리에 의해 유의적으로 AQP-3의 발현이 억제되었으며, 본 발명의 여주 추출물은 코르티손 처리에 의해 감소된 AQP-3의 발현을 농도의존적으로 회복시켰다. 또한, 30 ㎍/㎖의 농도부터 86.92%로 유의적인 AQP-3 회복 효과를 보였으며, 50 ㎍/㎖의 농도에서 최대 107.84% AQP-3의 발현이 촉진되는 효과를 나타내는 점을 확인하였다. 한편, 도 4는 여주 추출물의 코르티손 처리에 의해 감소된 아쿠아포린-3(AQP-3)의 발현 회복 효과를 확인한 결과이다.As a result of the experiment, as shown in Table 4 and Figure 4, the expression of AQP-3 was significantly suppressed by cortisone treatment, and the bitter melon extract of the present invention restored the expression of AQP-3 reduced by cortisone treatment in a concentration-dependent manner. I ordered it. In addition, it was confirmed that a significant AQP-3 recovery effect of 86.92% was observed at a concentration of 30 μg/ml, and that the expression of AQP-3 was promoted by up to 107.84% at a concentration of 50 μg/ml. Meanwhile, Figure 4 shows the results confirming the effect of recovering the expression of aquaporin-3 (AQP-3) reduced by cortisone treatment of bitter melon extract.
[실험예 5 : 여주 추출물의 filaggrin 발현 회복 효과][Experimental Example 5: Restoration effect of filaggrin expression of bitter melon extract]
본 실험에서는 여주 추출물의 스트레스에 인해 파괴된 피부 장벽 개선 효과를 확인하기 위해 피부 장벽 강화에 관여하는 인자인 필라그린(filaggrin)에 대한 영향을 확인하였다. In this experiment, in order to confirm the effect of the bitter melon extract on improving the skin barrier damaged by stress, the effect on filaggrin, a factor involved in strengthening the skin barrier, was confirmed.
각질형성세포를 6 웰 플레이트 (well plate)에 4 × 105 cell/well로 분주하고 37℃, 5% CO2 배양기에서 하루 동안 세포를 안정화시켰다. 코르티손(cortisone) 100 μM을 처리하여 스트레스 자극을 준 후, 상기 실시예 1에서 제조한 여주 추출물을 10, 30, 50 ㎍/㎖ 농도로 희석하여 각각 처리한 후 6시간 동안 반응시켰다. 배지를 제거하고 1 × PBS로 한번 세척한 후 스크래퍼로 긁어 세포를 수집하였다. 트리졸(Trizol)법을 이용하여 RNA를 분리하였다. Trizol 800 ㎕를 첨가하여 세포를 용해시키고 상온에서 5분 동안 반응시킨 후 클로로포름(chloroform) 200 ㎕를 첨가하여 볼텍싱(voltexing)한 다음 상온에서 5분 동안 반응시켰다. 12,000 rpm, 4℃로 15분 동안 원심분리하고 상등액만 새로운 튜브(tube)로 옮겨준 후 동량의 isopropanol을 첨가하여 1~3 번 섞어주고 상온에서 10분 동안 반응시켰다. 12,000 rpm, 4℃로 15분 동안 원심분리하고 상등액을 모두 제거한 후 차가운 70% 에탄올을 800 ㎕씩 첨가하여 세척하고 다시 12,000 rpm, 4℃로 15분 동안 원심분리하였다. 상층액을 완전히 제거하고 펠릿(pellet)이 투명해질 때까지 건조시킨 후 RNA 양에 따라 DEPC DW를 첨가하였다. Nanodrop을 이용하여 RNA를 정량하여 PCR을 수행한 후 2% 아가로오스 겔 (agarose gel)에서 전기영동하여 밴드를 검출하였다. 밴드는 Image J 1.47 software를 이용하여 수치화하였으며 베타-액틴(β-actin)의 발현을 기준으로 정량하였다. 한편, 아래 표 5는 코르티손 처리군 대비하여 필라그린(filaggrin)의 발현 회복율(%)을 계산하여 나타낸 것이다.Keratinocytes were distributed in a 6-well plate at 4 × 10 5 cells/well, and the cells were stabilized in an incubator at 37°C and 5% CO 2 for one day. After providing stress stimulation by treating with 100 μM of cortisone, the bitter melon extract prepared in Example 1 was diluted to concentrations of 10, 30, and 50 μg/ml, respectively, and reacted for 6 hours. The medium was removed, washed once with 1 × PBS, and cells were collected by scraping with a scraper. RNA was isolated using the Trizol method. Cells were lysed by adding 800 ㎕ of Trizol and reacted at room temperature for 5 minutes, then 200 ㎕ of chloroform was added, vortexed, and reacted at room temperature for 5 minutes. Centrifuge at 12,000 rpm and 4°C for 15 minutes, transfer only the supernatant to a new tube, add the same amount of isopropanol, mix 1 to 3 times, and react at room temperature for 10 minutes. It was centrifuged at 12,000 rpm and 4°C for 15 minutes, and all supernatants were removed, washed by adding 800 ㎕ of cold 70% ethanol, and centrifuged again at 12,000 rpm and 4°C for 15 minutes. The supernatant was completely removed, the pellet was dried until it became transparent, and DEPC DW was added according to the amount of RNA. RNA was quantified using Nanodrop, PCR was performed, and the band was detected by electrophoresis on a 2% agarose gel. The bands were quantified using Image J 1.47 software and quantified based on the expression of beta-actin. Meanwhile, Table 5 below shows the calculated recovery rate (%) of filaggrin expression compared to the cortisone treatment group.
(여주 추출물)Example 1 (㎍/㎖)
(Bitter melon extract)
실험 결과, 표 5 및 도 5에서 보듯이, 코르티손 처리에 의해 유의적으로 필라그린의 발현이 억제되었으며, 본 발명의 여주 추출물은 코르티손 처리에 의해 감소된 필라그린의 발현을 농도의존적으로 회복시켰다. 또한, 10 ㎍/㎖의 농도부터 93.03%로 높은 필라그린 촉진 효과를 보였으며, 50 ㎍/㎖의 농도에서 최대 220.99% 필라그린의 발현이 촉진되는 효과를 나타내는 점을 확인하였다. 한편, 도 5는 여주 추출물의 코르티손 처리에 의해 감소된 필라그린(filaggrin)의 발현 회복 효과를 확인한 결과이다. As a result of the experiment, as shown in Table 5 and Figure 5, the expression of filaggrin was significantly suppressed by cortisone treatment, and the bitter melon extract of the present invention restored the expression of filaggrin reduced by cortisone treatment in a concentration-dependent manner. In addition, it was confirmed that the effect of promoting filaggrin was as high as 93.03% at a concentration of 10 μg/ml, and that the expression of filaggrin was promoted by up to 220.99% at a concentration of 50 μg/ml. Meanwhile, Figure 5 shows the results confirming the effect of restoring the expression of filaggrin, which was reduced by cortisone treatment of bitter melon extract.
이와 같이, 본 발명의 여주 추출물은 스트레스 호르몬인 코르티솔의 생성 억제, 코르티솔 생성 조절 효소인 11β-HSD1의 발현 억제, 스트레스로 인해 감소된 피부 장벽 인자인 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin)의 발현 회복 효과를 통해 스트레스 완화 및 스트레스로 인해 무너진 피부 장벽 개선 효과를 발휘하는 점을 확인할 수 있었다.In this way, the bitter melon extract of the present invention inhibits the production of cortisol, a stress hormone, suppresses the expression of 11β-HSD1, an enzyme that regulates cortisol production, and inhibits aquaporin-3 (AQP-3) and filaggrin, which are skin barrier factors reduced due to stress. It was confirmed that the recovery effect of filaggrin relieves stress and improves the skin barrier damaged by stress.
Claims (4)
상기 추출물은 60~80%(v/v) 에탄올을 추출용매로 하여 상온에서 추출한 것으로, 코르티솔 생성 및 코르티솔 생성 조절 효소인 11β-HSD1의 발현을 억제하고, 스트레스로 인해 감소된 아쿠아포린-3(AQP-3) 및 필라그린(filaggrin)의 발현을 촉진하는 것을 특징으로 하는 피부 스트레스 개선 및 스트레스로 인한 피부 장벽 손상 개선용 화장료 조성물.
Contains bitter melon ( Momordica charantia ) ethanol extract as an active ingredient,
The extract was extracted at room temperature using 60-80% (v/v) ethanol as an extraction solvent, and inhibited the expression of 11β-HSD1, an enzyme that regulates cortisol production and cortisol production, and aquaporin-3 (aquaporin-3) reduced due to stress. A cosmetic composition for improving skin stress and improving skin barrier damage caused by stress, characterized in that it promotes the expression of AQP-3) and filaggrin.
상기 추출물은,
여주의 꽃, 잎, 열매, 줄기 또는 뿌리 중 선택되는 어느 하나 이상의 부위에서 얻은 추출물인 것을 특징으로 하는 화장료 조성물.According to paragraph 1,
The extract is,
A cosmetic composition characterized in that it is an extract obtained from any one or more parts selected from the flowers, leaves, fruits, stems, or roots of bitter melon.
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