In vitro evaluation systems for depigmentation and anti-wrinkle efficacy

안수미 ( Soo Mi Ahn ) 한국동물실험대체법학회 2008 한국동물실험대체법학회 학술대회집 Vol.2008 No.1

The cosmetics industry grows with the advancement of the society and culture. It is one field of typical fine chemical industry where the basic science and application technique of chemistry, biology, physiology and the pharmacy are applied together. The cosmetic products must have safety, efficacy, stability and user convenience. Recently the regulation against animal experiments is claimed in Europe and many in vitro methods are used and developed. In Korea, functional cosmetic fields are growing after cosmetic law was separated from pharmaceutical affairs law at June, 2000. Sunscreen, whitening and anti-wrinkle effectsare three categories in functional cosmetic law. Recently many cosmeceutical ingredients are developed which have a more biological activity than before in these categories. In this lecture I will review the detail of assessment method for evaluation of anti-wrinkle and depigmentaion agents in vitro. Collagen and MMP-1 are well known biological targets for evaluation of anti-wrinkle ingredients. Usually human dermal fibroblast (HDF) is used for these assays. The selected ingredients are evaluated in in vivo animal or human study. Initial evaluation of depigmenting properties should be performed on purified tyrosinase and other melanogenic enzymes, thereafter employing melanocyte cultures, cytotoxicity and effects on melanin synthesis should be assayed. All these evaluation methods have limits and methods which have higher relationships with human should be developed continuously with reliable biomarkers.

인체각질형성세포에서 Fructose 1,6-diphosphate의 자외선에 의해 유도되는 Cyclooxygenase-2 and Matrix Metalloproteinases의 발현억제기전

안수미 ( Soo Mi Ahn ),김지현 ( Ji Hyun Kim ),이병곤 ( Byeong Gon Lee ),이수환 ( Soo Hwan Lee ),장이섭 ( Ih Seoup Chang ) 대한화장품학회 2004 대한화장품학회지 Vol.30 No.2

자외선은 피부에 염증반응이나 광노화와 같은 다양한 반응을 야기시킨다고 알려져 있다. 특히 자외선에 의해 손상을 받은 피부는 콜라겐의 양이 감소되어 있는데, 이는 자외선에 의해 피부 내에서 콜라겐을 분해하는 효소(MMP, matrix metalloproteinases)의 양이 증가하기 때문이라고 알려져 왔다. 또한 자외선에 의해 피부에서 염증반응이 유발되는데, 이러한 반응은 프로스타글란딘이라는 물질에 의해 매개되며, 이 프로스타글란딘에 의해서도 MMP가 증가한다고 알려져 왔다. 본 연구에서는 6개월의 임상실험을 통해 광노화된 피부에서 주름형성억제효능이 뛰어난 FDP (fructose 1.6-diphosphate)의 작용기전을 인체각질형성 세포를 이용하여 연구하였다. 인체각질형성세포에 자외선을 조사할 경우 프로스타글란딘, COX-2 (cyclooxygenase-2), MMPs의 활성이 증가하는 것을 확인하였며, 이는 FDP의 처리에 의해 감소되었다. 이러한 효과는 자외선에 의해 인체각질형성세포에서 발생하는 신호전달과정을 억제함으로써 일어나는 효과임이 증명되었다. 따라서, FDP는 자외선에 의해 일어나는 세포 내 신호전달과정을 억제하며, 이로 인해 야기되는 프로스타글란딘, COX-2, MMPs의 증가를 억제함으로써 피부의 광노화를 억제할 수 있는 원료로 여겨진다. UV radiation exerts various influences in the skin, including photoaging and inflammation (1). The MMPs (Matrix metalloproteinases), which are induced by UV irradiation, can degrade matrix proteins, and these results in a collagen deficiency in photodamaged skin that leads to skin wrinkling. It has been known that the production of PGE₂ stimulates MMPs expression, and inhibits procollagen (2). Thus, it is possible that the induction of MMPs and the inhibition of matrix protein synthesis by UV -induced PGE₂ may play some role in UV-induced collagen deficiency in photoaged skin. Fructose-1,6-diphosphate (FDP), a glycolytic metabolite, is reported to have cytoprotective effects against ischemia and postischemic reperfusion injury of brain and heart, presumably by augmenting anaerobic carbohydrate metabolism (3). And also, FDP significantly prevent skin aging by decreasing facial winkle compared with vehicle alone after 6 months of use. We studied the mechanism of anti-aging effect of FDP on UVB-irradiated HaCaT keratinocyte model. FDP has protective role in UVB injured keratinocyte by attenuating prostaglandin E₂ (PGE₂) production and COX-2 expression. And FDP also suppressed UVB-induced MMP-2 expression. Further, to delineate the inhibition of UVB-induced COX-2 and MMPs expression with cell signaling pathways, treatment of FDP to HaCaT keratinocytes resulted in marked inhibition of UVB-induced phosphorylation of ERK1/2, JNK. It also prevents UV induced NFB translocation, which are activated by cellular inflammatory signal. Our results indicate that FDP has protecting effects in UV-injured skin aging by decreasing UVB-induced COX-2 and MMPs expression, which are possibly through blocking UVB-induced signal cascades.

자산관리과정과 부부의 역할구조

安秀美(Ahn Soo Mi),李基榮(Lee Ki Young) 가정과삶의질학회 1998 가정과삶의질연구 Vol.16 No.4

Adamantyl Benzamide 유도체의 미백효과

백흥수 ( Heung Soo Baek ),안수미 ( Soo Mi Ahn ),우병영 ( Byoung Young Woo ),조영석 ( Young Seok Cho ),최수정 ( Soo Jeong Choi ),노호식 ( Ho Sik Rho ),변경희 ( Kyoung Hee Byoun ),신송석 ( Song Seok Shin ),박영호 ( Young Ho Park 대한화장품학회 2013 대한화장품학회지 Vol.39 No.2

Polyhydroxylated benzamide 유도체의 구조변화에 따른 미백효과의 상관관계를 고찰하였다. Adamantyl benzamide 유도체에서 B ring 부분의 치환기가 catechol (3,4-dihydroxyphenyl)인 경우 우수한 멜라닌 생성 저해활성을 보였으나, mono-hydroxyphenyl (3-OH 또는 4-OH)이거나 3,4-dimethoxyphenyl인 경우에 그 활성이 감소하거나 없어졌다. 따라서 catechol unit이 멜라닌 생성 저해에 중요한 인자임을 알 수 있었다. 그리고 A-ring부분의 2-OH의 존재여부는 활성에 큰 영향을 주지는 않았고, A-ring과 B-ring을 연결하는 탄소사슬의 길이 역시 멜라닌 생성저해에 큰 영향을 주는 요소는 아니었다. The structure activity relationship of polyhydroxylated benzamide derivatives for whitening effects was examined. The adamantyl benzamide derivatives with catechol (3,4-dihydroxyphenyl) of B-ring part showed good anti- melanogenesis activity, but the inhibitory activity of mono-hydroxyphenyl (3-OH or 4-OH) or 3,4-dimethoxyphenyl substituted derivatives was decreased or lost. Therefore the catechol unit was appeared to be the crucial factor for the inhibition of melanogenesis. And the existence of 2-OH of A-ring part had minor influence on the activity, the length of carbon chain between A-ring and B-ring was also not the major factor for the anti-melanogenesis activity.

연구논문 : 기능성 화장품 소재개발을 위한 in vitro 평가법

이진영 ( Jin Young Lee ),박원석 ( Won Seok Park ),손의동 ( Eui Dong Son ),안수미 ( Soo Mi Ahn ),황재성 ( Jae Sung Hwang ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.1

The cosmetics industry grows with the advancement of the society and culture. The cosmetic products must have safety, efficacy, stability and user convenience. In Korea, functional cosmetic fields are growing after cosmetic law was separated from pharmaceutical affairs law at June, 2000. Sunscreen, whitening and anti-wrinkle effects are three categories in functional cosmetic law. Though not included in functional cosmetics, anti-hair loss and hair growth products are also important in cosmetics and cosmeceuticals. Recently the regulation against animal experiments is claimed in Europe and many in vitro methods are used and developed. In vitro evaluation methods for whitening, anti-wrinkle and hair growth ingredients are introduced in this paper. Collagen and MMP-1 are well known biological targets for evaluation of anti-wrinkle ingredients. For the whitening effect, the activity of purified tyrosinase and other melanogenic enzymes are tested, thereafter employing melanocyte cultures, cytotoxicity and effects on melanin synthesis should be assayed. In vitro dermal papillar cell culture and hair organ culture are generally used for the screening of hair growth ingredients. Co-culture systems and reconstructed skin seem to be reliable methods to screen the capability of new agents compared to monolayer cell culture system. But all these evaluation methods have limits, and methods which have higher relationships with human should be developed continuously with reliable biomarkers.

포도나무가지 추출물의 프리라디칼 소거 작용 및 염증 발현 매개인자 생성 억제 효과

허선경(Sun Kyung Hur),김선숙(Sun Sook Kim),허연희(Yeon Hoi Heo),안수미(Soo Mi Ahn),이병곤(Byeong Gon Lee),이상국(Sang Kook Lee) 한국응용약물학회 2001 Biomolecules & Therapeutics(구 응용약물학회지) Vol.9 No.3

N/A Free radical scavengers or quenching agents for reactive oxygen species (ROS) present in consumable fruits, vegetables, and beverages have received considerable attention as potential antioxidants, and thus uses for treatment of several human diseases. In this study, grapevine shoot extract (GSE) containing high concentration of resveratrol and viniferine was evaluated for antioxidant potential and inhibition of pro-inflammatory mediator production. Utilizing 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and xanthine oxidase (XOD) inhibition assay the GSE showed inhibitory effects of DPPH radical scavenging and XOD activity with the IC_(50) values of 34.5 and 155 ㎍/㎖, respectively. In addition, GSE also exhibited the inhibition of prostaglandin E₂ (PGE₂) and nitric oxide (NO) production in lipopolysaccharide (LPS)-induced mouse macrophage RAW264.7 cells with the IC_(50) value of 6.4 and 14.5 ㎍/㎖, respectively. This result suggests that grapevine shoot extract has the potential activity as a natural antioxidant or antiinflammatory agent.

기능성 화장품 소재개발을 위한 in vitro 평가법

이진영 ( Jin Young Lee ),박원석 ( Won Seok Park ),손의동 ( Eui Dong Son ),안수미 ( Soo Mi Ahn ),황재성 ( Jae Sung Hwang ) 한국동물실험대체법학회 2007 동물실험대체법학회지 Vol.1 No.1

The cosmetics industry grows with the advancement of the society and culture. The cosmetic products must have safety, efficacy, stability and user convenience. In Korea, functional cosmetic fields are growing after cosmetic law was separated from pharmaceutical affairs law at June, 2000. Sunscreen, whitening and anti-wrinkle effects are three categories in functional cosmetic law. Though not included in functional cosmetics, anti-hair loss and hair growth products are also important in cosmetics and cosmeceuticals. Recently the regulation against animal experiments is claimed in Europe and many in vitro methods are used and developed. In vitro evaluation methods for whitening, anti-wrinkle and hair growth ingredients are introduced in this paper. Collagen and MMP-1 are well known biological targets for evaluation of anti-wrinkle ingredients. For the whitening effect, the activity of purified tyrosinase and other melanogenic enzymes are tested, thereafter employing melanocyte cultures, cytotoxicity and effects on melanin synthesis should be assayed. In vitro dermal papillar cell culture and hair organ culture are generally used for the screening of hair growth ingredients. Co-culture systems and reconstructed skin seem to be reliable methods to screen the capability of new agents compared to monolayer cell culture system. But all these evaluation methods have limits, and methods which have higher relationships with human should be developed continuously with reliable biomarkers.

pH 감응형 나노입자를 이용한 멜라닌 합성저해 연구

박주영 ( Ju Young Park ),최현정 ( Hyun-jung Choi ),심종원 ( Jong Won Shim ),안수미 ( Soo Mi Ahn ),김준오 ( Junoh Kim ),장이섭 ( Ih-seop Chang ) 대한화장품학회 2004 대한화장품학회지 Vol.30 No.1

내형질 세망 조직에서 N-글리코실레이션 과정의 초기 단계를 차단하면 멜라닌 생 합성의 주 효소인 티로시나제의 활성이 저해된다. 본 연구에서는 in vitro 환경에서 N-글리코실레이션 저해제의 활성을 증가시키고자 전달체로 pH 감응성을 갖는 나노크기의 지질구조체를 제조하고 이를 평가하였다. 이 pH 감응성 지질구조체 Melexsome은 일반적인 지질성분인 포스포리피드와 콜레스테롤 기반의 지질안정 성분으로 구성되며, 통상적인 리포좀 제조법에 따라 제조되었다. 글리코실레이션 저해 성분물질을 포집시킨 Melel[some의 효과는 EndoH & PNGaseF 분해와 western blotting 방법에 의해 평가하였고, 멜라닌 합성량 또한 측정 되었다. 이 결과, pH 감응성을 갖도록 제조된 Melexsome이 N-글리코실레이션 저해제의 효능을 효과적으로 증진시킴을 알 수 있었다. 또한, 공초점 주사 현미경에 의한 세포관찰 결과에 따르면 Melexsome은 여타의 전달체에 비하여 세포질 내에 보다 효과적으로 전달되는 것으로 보여지며, 따라서 이같은 양친성 지실성분 기반의 pH 감응성 나노 전달체는 N-글리코실레이션 저해제의 전달 시스템으로서 미백 화장료 제품이 가져야 하는 침착된 색소에 의해 어두워진 피부톤의 개선 효과를 극대화 시키는데 적합하다고 여겨진다. Inhibition of the early N-glycosylation process in the endoplasmic reticulum prevents the activation of tyrosinase, a key enzyme for melanin biosynthesis. This work aims at evaluating the increased activity of N-glycosylation inhibitors in vitro b, employing a nano-sized pH-sensitive liposome as a delivery carrier. Melexsome, a pH-sensitive nano carrier loaded with glycosylation inhibitos, was prepared by the hydration method with phospholipids and cholresterol-based amphiphiles. Inhibitory effects of Melexsome on the N-glycosylation process were evaluated by EndoH & PNGaseF digestion and the western blotting. Melanin synthesis was also monitored after treatment with Melexsome Interestingly, Melexsome effectively increased the efficacy of N-glycosylation inhibitors. Melexsome was also much more efficiently translocated into the cytoplasm as observed in CLSM. These results demonstrated that the amphiphilic lipid-based pH-sensitive nano-carriers could be, used as an efficient delivery system for N-glycosylation inhibitor to enhance the effects of skin whitening cosmetics.

Coenzume Q10의 멜라닌 생성억제 효과

황재성 ( Jae-sung Hwang ),박원만 ( Won- Man Park ),안수미 ( Soo-mi Ahn ),강병영 ( Beyong-young Kang ),이병곤 ( Byeong-gon Lee ),심영철 ( Young- Chul Sim ) 대한화장품학회 2000 대한화장품학회지 Vol.26 No.1

Coenzyme Q10(CoQ10)은 피부를 포함한 모든 생체조직에 존재하는 널리 알려진 조효소이다. 전자전달에 관여하는 퀴논링은 세포에서 에너지를 생성하기 위한 매우 중요한 기능을 가지고 있다. CoQ10 은 피부에서 항산화제로서 연구되어 왔으며, 최근 외용제로써 노화억제와 주름개선작용에 대해 보고된 바 있다. 이런 보고들은 CoQ10 이 항산화제로서 산화-환원작용을 통해 피부의 방어기능에 중요한 역할을 한다는 점을 시사하며, 일반적으로 산화-환원작용은 피부에서 흑화과정의 조절에도 많은 영향을 미친다. 따라서 본 연구자들은 CoQ10 이 피부의 색소조절기능이 있는지 알아보고자 하였다. 인체 정상 색소세포에 CoQ10을 0.05-0.5 mM 처리한 결과 0.5, 0.25mM 에서 멜라닌의 생함성을 약 50% 저해하였으며 이는 알려진 미백제인 Kojic acid 나 vitamin C 와 유사한 수준이었다. 또한, CoQ10 은 인체 정상 색소세포에서 자외선이나 세포내 cAMP 증가 유도물질에 의한 멜라닌 생성을 억제하였다. 그러나 tyrosinase inhibitor 인 kojic acid 와는 달리, in vitro tyrosine hydroxylase 의 억제효과는 보이지 않았다. CoQ10을 자외선으로 tanning 을 유도시킨 brown guinea pig 에 4 주간 도포하고 육안 및 chromameter 를 이용하여 미백효과를 측정한 결과, vehicle 처리군에 비해 미백효과가 있음을 확인할 수 있었다. 이상의 결과에서 coenzyme Q10 은 in vitro 및 in vivo 에서 미백효과를 지닌 물질임을 확인할 수 있었다. Coenzyme Q10 is found in all tissues including skin and it is the well-known coenzyme for mitochondrial enzymes. The electron and proton transfer functions of the quinone ring are of fundamental importance for the oxidative phosphorylation pathway to generate energy in the cells. Coenzyme Q10 has been studied as a potent antioxidant molecule in the skin. It is involved in the skin’s response to UVR irradiation. The concentration of this antioxidant in UVR exposed skin is higher than in non-exposed skin. However, recent studies have also shown that coenzyme Q10 is one of the first antioxidants to be depleted when skin is UVR-irradiated. This indicates that coenzyme Q10 is primarily involved in defense mechanisms of the skin. Therefore, we questioned whether coen2yme Q10 shows regulatory effect of melanogenesis. Here we report that coenzyme Q10 inhibits melanin neosynthesis of normal human melanocytes grown in culture, and lightens UVB-induced hyperpigmentation of the guinea pig skin in vivo. We treated human melanocytes with 0.05mM to 0.5mM of coenzyme Q10 for a total of two days. This inhibited melanin neosynthesis of cultured human melanocytes dose-dependently. The inhibitory effects of coenzyme Q10 was as effective as kojic acid or vitamin C on cultured human melanocytes. CoQlO didn’t have direct inhibitory effect on tyrosinase activity in in vitro tyrosine hydroxylase activity. To further clarify the effect of coenzyme Q10 on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brownish guinea pigs. The UVB intensity was 500mJ/cm² and the total energy dose was 1,500 mJ/cm². The animals were exposed to UVB radiation one times a week for three consecutive weeks. Coenzyme Q10, kojic acid, Arbutin, vitamin C(1% in vehicle) or vehicle alone as a control were then topically applied daily to the hyperpigmented areas twelve times per week for four successive weeks. The lightening effect was evaluated by visual scoring, chromameter and immunohistochemistry. Coenzyme Q10 had lightening effect on the UVB-induced hyperpigmentation without any other side effects, whereas another compounds showed weak lightening efficacies. Therefore, these results suggest that coenzyme Q10 may be useful for solving physiological hyperpigmenting problems for cosmetic purposes.

바다 - Ⅱ DBMS를 위한 연결과 접근 권한의 설계 및 구현

곽세진(Se Jin Kwak),백진아(Jin A Baek),박성철(Seong Cheol Park),차명훈(Myung Hoon Cha),안수미(Soo Mi Ahn),박영철(Young Chul Park),이미영(Mi Young Lee) 한국정보과학회 1996 한국정보과학회 학술발표논문집 Vol.23 No.2A

본 논문은 메시지 큐 방식에서 소켓 방식으로 전환한 바다-Ⅱ DBMS의 연결과정을 소개하고 관계형 데이타베이스 관리시스템의 데이타베이스에 대한 권한 관리와 데이타베이스 내부의 객체에 대한 권한을 관리하기 위한 방법을 SQL의 국제표준, Oracle, Informix, Sybase등을 참조하여 바다-Ⅱ DBMS에 구현한 내용을 기술한다. 구현 내용은 데이타베이스 수준의 권한과 데이타베이스 내부 객체 수준의 권한으로 나뉘어지며 데이타베이스 수준의 권한으로는 CONNECT, RESOURCE, DBA 권한이 있고, 객체 수준의 권한으로는 테이블, 뷰, 색인, 도메인, 문자집합, 저장함수에 관한 권한이 있다.