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1 Korean Society for Biotechnology and Bioengineering Journal 32(1): 9-15 (2017) ISSN / eissn Research Paper HaCaT 세포주에서캐모마일 (Matricaria chamomilla) 추출물의항병원성및항염효과에관한연구 임은경 1, 김근태 1, 김보민 1, 김은지 1, 김상용 2, 한남규 3, 하재순 3, 김영민 1 * Study of Anti-microbial Activities and Anti-inflammatory Effects of Chamomile (Matricaria chamomilla) Extracts in HaCaT cells Eun Gyeong Lim 1, Guen Tae Kim 1, Bo Min Kim 1, Eun Ji Kim 1, Sang-Yong Kim 2, Nam Kyu Han 3, Jae Sun Ha 3, and Young Min Kim 1 * Received: 2 January 2017 / Revised: 31 January 2017 / Accepted: 2 February The Korean Society for Biotechnology and Bioengineering Abstract: Chamomile (Matricaria chamomilla), a member of the Asteraceae family, is a well-known for medicinal plant and can be found in India and Europe. Chamomile is an effective sedative and various medical effects. But, the effects of acne treatment by chamomile were not investigated. Therefore, we assessed the anti-oxidant effects, anti-microbial activity and anti-inflammatory effects by chamomile extracts in HaCaT keratinocyte cells. Anti-oxidant effects of chamomile extracts were investigated by DPPH assay. Also, results of MTT assay was demonstrated that chamomile extracts did not have a cytotoxic effect in HaCaT cells. To assess the antimicrobial activity, we determined formation of inhibition zone of Propionibacterium acnes by extracts from chamomile. Tumor necrosis factor-α (TNF-α) induces production of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6 and IL- 8 and expression of COX-2. Chamomile extracts could inhibit 1 한남대학교생명나노과학대학생명시스템과학과 1 Department of Biological Sciences and Biotechnology, College of Life Science and Nano Technology, Hannam University, Daejeon 34054, Korea Tel: , Fax : kym@hnu.kr 2 신안산대학교식품생명과학과 2 Department of Food Science & Bio Technology, Shinansan University, Ansan, Korea 3 닥터하스킨주식회사 3 Dr. HASKIN Co., Ltd., Korea TNF-α-induced mrna expression levels of IL-1β, IL-6, IL-8 and COX-2 gene. These results demonstrated the possibility of chamomile for prevention and treatment of skin inflammatory diseases such as acne. Keywords: Chamomile, Anti-microbial activity, Anti-inflammatory effects, P.acnes, HaCaT cells 1. INTRODUCTION 여드름은피지선주변에서발생하는만성염증질환으로면포, 구진, 농포, 낭종, 결절등의다양한병변을동반하며, 잘못된관리시흉터등의후유증을남긴다 [1]. 여드름의발생에는복합적인요인이작용하는것으로알려져있으며, 일반적으로피지선에서의과도한피지분비, 호르몬불균형, 유전, 비정상적인모낭내각화현상및 Propionibacterium acnes 의증식에의한염증반응등에의하여발생한다 [2]. 여드름은주로사춘기청소년층에서관찰되지만최근육식위주의식습관변화와환경호르몬등에의하여 25 세이상의청년층에서나타나는성인여드름이증가하는추세다 [3]. 일반적인여드름치료법은국소요법과전신요법으로나뉘며국소요법으로는 erythromycin, clindamycin 을포함한국소항생제와벤조일과산화물 (Benzoyl peroxide), tretinoin, adapalene 등의국소레티노이드제와레이저치료, 압출등의방법이있으며, 전신요법으로는호르몬제 ( 경구피임제, spironolactone) 등이있다 [1,3]. 하지만이러한방법들은피부자극이심하고항생

2 10 Korean Society for Biotechnology and Bioengineering Journal 32(1): 9-15 (2017) 제및스테로이드제에대한내성및부작용등의문제점이발생하기때문에최근에는피부에자극이적고안전한천연추출물유래여드름치료물질개발에대한관심이높아지는추세이다 [3]. 실제로최근소비자들의소비추세가자연지향적이고환경친화적으로변해감에따라화장품에들어가는유효성분도화학물질보다는식물유래의천연물을선호하고있으며, 천연물의유용성을기반으로하여여러가지형태로화장품에배합되어사용되고있다 [4,5]. 캐모마일 (Matricaria chamomilla) 은인도와유럽이원산지인국화과다년생식물로황색의꽃이피는다이어즈캐모마일과백색의꽃이피는로만캐모마일로나뉜다 [6]. 캐모마일은피부살균효과가있어서유럽에서는미용을위한목욕제로쓰이며, 감기예방, 불면증해소등의효과가알려져있기때문에주로차로음용되어왔다 [6]. 캐모마일의꽃에는 terpenes, flavonoids, coumarin, calicacid 등의성분이함유되어있으며, 꽃을증류추출하여얻은에션셜오일은푸른빛을띠고 α-bisabolol, chamazulene, bisabolol oxide 등의성분이포함되어있어염증을가라앉히는데효과적이다 [6-8]. 또한캐모마일은진정작용, 항당뇨효과, 항관절염효과등의다양한약물효능이있는것으로보고되었다 [9-11]. 하지만아직까지여드름과같은특정피부염증질환에대한캐모마일의과학적효능에대한연구는미미한실정이다. 따라서본연구에서는캐모마일추출물의항산화효과와여드름유발균인 P.acnes 균에대한항균효과, 피부각질형성세포에서의항염증효과를확인함으로써여드름개선화장료로서캐모마일의가능성을확인하고자하였다. 2. MATERIALS AND METHOD 2.1. 실험재료및기기실험에사용된 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH) 와 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphnyl-2h-tetrazolium bromide (MTT) 는 Sigma-Aldrich Co. (St. Louis, MO, USA) 에서구입하였으며에탄올, 클로로폼, 이소프로판올등의시약은시판특급시약을사용하였다. 시료의흡광도는 ELISA microplate reader (Model 680, Bio-Rad Laboratories, Inc., Tokyo, Japan) 를사용하여측정하였다 캐모마일추출물제조본실험에서사용된캐모마일은경상남도진주시에위치한허브줌농원에서구입하였으며, 캐모마일꽃 1 Kg 에증류수 8 L 를가하여 2 h 동안증류추출하였다. 이러한방법을통하여얻어진추출물의상층부분에생성된캐모마일오일과하단부분의캐모마일플로럴워터는분리하여빛을차단한상 태에서상온보관하였다. 실험에사용된캐모마일워터는 1 g/ml 의농도로보관하였고, 캐모마일오일은 30 mg/ml 의농도로캐모마일플로럴워터에희석하여보관한후실험에사용하였다 균주와세포주배양 HaCaT 세포는 Dr. Fusenig (German Cancer Research Center, DKFZ, Heidelberg, Germany) 로부터분양받았으며, Fibroblast 세포는 American Type Culture Collection (ATCC, Rockville, MD, USA) 에서분양받아사용하였다. 세포는 10% FBS 와 1% antibiotics 가포함된 DMEM media (Hyclone, Laboratris Inc., Logan, UT, USA) 를사용하여 5% CO 2, 37 o C 조건하에서배양하였다. 매 48 h 마다 Trypsin-EDTA (Hyclcone, Laboratories Inc., Logan, UT, USA) 를이용하여세포를부유상태로만든다음세포를 cells/ml 로분주하고계대배양하였다. 여드름유발균주인 Propionibacterium acnes (P. acnes KCTC 3314) 는한국생명공학연구원생물자원센터에서분양받아사용하였으며, Reinforced Clostridial Agar (MB-R1096, MBcell, Korea) 를사용하여매 72 h 마다 37 o C, anaerobic 조건하에서배양하였다 (Table 1) DPPH assay 캐모마일워터및오일의항산화효능은 DPPH 법을통하여확인하였다. 시료로사용된캐모마일워터는 1 mg/ml, 캐모마일오일은 15, 30, 60 μg/ml의농도가되도록 94.9% 에탄올에희석하여사용하였다. 94.9% 에탄올에용해시킨 0.2 mm DPPH 용액 1 ml과시료 1 ml을혼합하고빛을차단하여 30 min 동안반응시킨후 microplate reader를이용하여 517 nm 의파장에서흡광도를측정하였다. blank로는시료대신증류수를첨가하였고, Vitamin C를농도별로제조하여사전실험을진행한후에적정농도를양성대조군으로사용하였다. 자유라디칼소거능력은아래식을이용하여계산하였다. DPPH free radical scavenging rate (%) A blank A exp = 100 A blank 2.5. 세포생존율측정세포생존율은 MTT assay 법을통하여측정하였다. 12 well plate 에 HaCaT 세포와 fibroblast 세포를각각 cells/ml 로분주하고 24 h 동안안정화시킨후최종농도가캐모마일워터는 1 mg/ml, 오일은 30 μg/ml 의농도가되도록처리하여 24, 48 h 동안 5% CO 2, 37 o C 조건하에서배양하였다. 염증반응유도군에서는 TNF-α 를 1 h 전처리하였다. 배양이끝난후 5 mg/ml 3-(4,5-dimethyl-2 thiazolyl)-2,5-diphnyl-2h-tetra- Table 1. Culture condition of microorganisms for antimicrobial activity test Strains Media Temp ( o C) Propionibacterium acnes KCTC 3314 Reinforced Clostridial agar 37

3 HaCaT 세포주에서캐모마일 (Matricaria chamomilla) 추출물의항병원성및항염효과에관한연구 11 zolium bromide (MTT, Sigma-Aldrich Co.) solution 을각 well 당 20 μl 씩첨가하여 1 h 동안 CO 2 incubator 에서반응시켰다. MTT solution 이포함된 media 를제거하고 PBS washing 후 PBS 를완전히제거하였다. 각 well 당 DMSO 150 μl 을첨가하여생성된 formazan 을모두녹인다음 96 well plate 에 100 μl 씩옮긴후 ELISA microplate reader (Bio-Rad model 680, Bio-Red Laboratories Inc., Tokyo, Japan) 로 595 nm 에서흡광도를측정하였다. 측정은모두 3 번이상하였으며, 이에따른평균값과표준오차는 Microsoft Excel program 을사용하여분석하였다 생육환저해환시험 (Clear Zone Test) 여드름유발균에대한캐모마일워터및오일의항균활성측정은 clear zone test 를통해실시되었다. 여드름유발균인 P. acnes 를액체배지에접종하여 37 o C 에서 18 시간씩 2 회계대배양하여활성화시킨후실험에사용하였다. P.acnes 균을 1 백금이취하여평판배지에도말한후시료 25 μl 을완전히흡수시킨직경 6 mm 의멸균 paper disc 를올려놓고 37 o C, anaerobic 조건하에서 24 시간동안배양하였다. 배양후 disc 주위에생성된생육저해환 (clear zone) 크기 (mm) 를측정하여각시료의항균력을측정하였다. 음성대조군으로는아무것도처리하지않은 paper disc 와증류수를동량흡수시킨 paper disc 를사용하였으며, 양성대조군으로는항생제인 Gentamycin(10 μg/ml) 을동량흡수시킨 paper disc 를사용하였다 Total RNA 추출및 cdna 합성 6 well plate 에 HaCaT 세포를각각 cells/ml 로분주하고 24 h 동안안정화시킨후최종농도가캐모마일워터는 1 mg/ ml, 오일은 30 μg/ml 의농도가되도록처리하여 24, 48 h 동안 5% CO 2, 37 o C 조건하에서배양하였다. 염증반응유도군에서는 TNF-α 를 1 h 전처리하였다. 배양후 media 를모두제거한뒤 PBS 로 1 회 washing 후 RiboEX Total RNA Isolation Solution (GeneAll Biotechnology, Seoul, Republic of Korea) 을가하여세포를용해한후클로로폼, 이소프로판올, 70% 에탄올을각각첨가하는과정을통하여 total RNA 를추출하였다. cdna 합성은 DiaStar TM RT kit (SolGent, Seoul, Korea) 를이용하여진행하였다. total RNA 1 μl 와 oligo(dt)20 primer 9 μl 를혼합한뒤 65 o C 에서 5 min 동안반응시킨후 ice 에넣어반응을중단하였다. 상기과정을통하여 oligo(dt) primer 가붙은 mrna 에 8 mm DTT 1 μl, RNase (200 U/μL) 1 μl, 10 mm 의 dntp 가포함된 5X RT reaction buffer 4 μl, DEPC- DW 를첨가한후 40 o C 에서 1 h, 95 o C 에서 5 min 동안반응시킴으로써 cdna 를합성하였다 역전사중합효소연쇄반응 (Reverse Transcription Polymerase Chain Reaction, RT-PCR) PCR은 DNA polymerase와 buffer, dntp, tracking dye가포함된 2X Taq PCR Smart mix 1 (SolGent, Seoul, Korea) 를이용하여실시하였다. PCR 증폭은 95 o C에서 10 min 동안 pre-denaturation 하고, denaturation은 95 o C에서 30 sec, annealing은 60 o C에서 30 sec, extension은 72 o C에서 30 sec의조건으로 35 cycles 반응시켰으며 72 o C에서 10 min 동안안정화시켰다. IL-1β, IL-6, IL-8, COX-2, actin의 specific primer sequences는 Table 2와같다 통계처리통계프로그램인 SPSS 22.0 을사용하였고실험설계에대한분산분석은일원배치분산분석 (One-way ANOVA) 과독립표본 t-test (independent sample t-test) 를실시하여검정하였다. 각자료는 3 번이상의반복된실험을통하여얻어진결과로검정하였고 p<0.05 인경우통계적으로유의한것으로판정하였다. 3. RESULT AND DISCUSSION 3.1. 캐모마일추출물의항산화효과캐모마일워터및오일의항산화효과를측정하기위하여 DPPH assay 를실시하였다. DPPH 법은시료의환원력을측정하는실험방법으로시료가 DPPH 의자유라디칼에전자를공여함으로써발생하는자유라디칼소거능력을측정한다 [12]. 캐모마일워터및오일의항산화효과를측정한결과, 캐모마일워터는 13.3%, 캐모마일오일은 15 μg/ml 의농도에서 11.74%, 30 μg/ml 의농도에서 22.75%, 60 μg/ml 에서 Table 2. Specific primer sequences for the amplification of target gene Gene Primer sequence (5'-3') Amplification size (bp) Annealing temperature ( o C) IL-1β For: GGAGAATGACCTGAGCACCT Rev: GGAGGTGGAGAGCTTTCAGT IL-6 For: AGTCCTGATCCAGTTCCTGC Rev: AAGCTGCGCAGAATGAGATG IL-8 For: TGAGCATCTACGGTTTGCTG Rev: TGCTTGTCTGGAACAACTGC COX-2 For: TGAGCATCTACGGTTTGCTG Rev: TGCTTGTCTGGAACAACTGC Actin For: GGACTTCGAGCAAGAGATGG Rev: AGCACTGTGTTGGCGTACAG

Antioxidative effects were measured by DPPH assay. The statistical analysis of the data was carried out by use of a one-way ANOVA (p< 0.05, each experiment, n=3). 38.

4 12 Korean Society for Biotechnology and Bioengineering Journal 32(1): 9-15 (2017) 비교적높은항산화효과를지닌것으로볼수있다. 선행연구에의하면캐모마일에탄올추출물의항산화효과는 75.4% 로라벤더, 페퍼민트와같은기타허브류보다높은것으로보고된바있으며, 이러한결과는본실험의항산화효과보다높은수치이지만이는추출용매및방법에의한차이인것으로생각된다 [13]. Fig. 1. Free radical scavenging ability of chamomile extracts. Antioxidative effects were measured by DPPH assay. The statistical analysis of the data was carried out by use of a one-way ANOVA (p< 0.05, each experiment, n=3) % 의자유라디칼소거능이관찰되어항산화효과가있음을확인하였다 (Fig. 1). 이는양성대조군인 vitamin c 의항산화효과가 37.23% 인것과비교했을때, 캐모마일오일이 3.2. 캐모마일추출물의세포독성검사피부표피층을구성하는피부각질형성세포 (HaCaT) 는피부의보호막역할을하는각질을형성하며다양한자극에반응하여여러사이토카인 (cytokine) 을분비함으로써염증반응및면역반응에관여한다 [14,15]. 피부각질형성세포가자극을받으면 tumor necrosis factor-α (TNF-α) 를분비하여다른사이토카인의분비를촉진함으로써염증을유도하는것으로알려져있다 [15]. 따라서본연구에서는피부를구성하고있는피부각질형성세포 (HaCaT) 및섬유아세포 (fibroblast) 에대한캐모마일추출물의세포독성여부를확인하기위하여 MTT assay 를실시하였다. 또한 TNF-α 로여드름성염증반응을유도한후 MTT assay 를실시함으로써여드름성염증유발피부에서캐모마일추출물의세포독성여부를확인하고자하였다. 캐모마일워터및오일을 24 h 및 48 h 동안처리한 Fig. 2. Cytotoxic Effects of Chamomile extract in HaCaT cells. Cell viability was measured by MTT assay. (a and c) Cells were treated with chamomile water (C.W) and oil (C.O) for 24 h and 48 h. (b and d) Cells were pre-treated with 10 ng/ml TNF-α for 1 h followed by treatment with chamomile water and oil for 24 h and 48 h. The statistical analysis of the data was carried out by use of a t-test. N.S. is not significant (each experiment, n=3).

5 HaCaT 세포주에서캐모마일 (Matricaria chamomilla) 추출물의항병원성및항염효과에관한연구 13 Table 3. Antimicrobial activities of chamomile extracts Diameter of Clear Zone (mm) Strains None D.W Gentamycin Chamomile Water Chamomile Oil P. acnes KCTC3314 ND a) ND 28.0±1.0 b) ND 10±0.0 b) a) ND, not detected, b) Inhibition Zone, The results are expressed as mean±sd of triplicates 후에세포생존율을측정한결과, Fig. 2 에나타난바와같이염증을유발한 24 h 처리군의경우, TNF-α 처리시세포생존율이 90.9%, 캐모마일워터처리군에서 96.8%, 캐모마일오일처리군에서 98% 로나타났으나통계적으로유의하지않았다 (Not Significant). 또한, 염증을유발한 48 h 처리군에서도 TNF-α 처리시세포생존율이 86.6% 로감소하는것으로나타났으나통계적으로유의한결과는아니었으며, 캐모마일워터처리군에서 96%, 캐모마일오일처리군에서 100.9% 로세포생존율이다시증가하는것을확인하였다. 따라서본실험을통하여캐모마일추출물이피부각질형성세포및섬유아세포에독성이없는것을확인하였다 캐모마일추출물의항균효과과각질화및과다한피지분비와같은다양한요인에의하여모공이막히면혐기성세균으로알려진 P.acnes 의모공내증식이활발하게진행된다. P.acnes 는지질분해요소인 lipase 를분비함으로써피지의주성분인 triglyceride 를분해하여 free fatty acid 를생성하는것으로알려져있으며, 생성된 free fatty acid 는모낭벽의상피세포를자극하여과각질화및피지분비를더욱더악화시켜염증을유발한다 [16]. 따라서본연구에서는여드름유발균인 P.acnes 균에캐모마일워터및오일을처리하여캐모마일의항균력을측정하고자하였다. 그결과 Table 3 과같이, 음성대조군인아무것도처리하지않은군과증류수를처리한군에서는 P.acnes 균에대한항균효과가나타나지않았으며, 양성대조군인 Gentamycin 처리군에서는 28.0±1.0 mm 의생육저해환이형성되었다. 또한캐모마일워터처리군에서는생육저해환이형성되지않았으나캐모마일오일처리군에서 10±0.0 mm 의생육저해환이형성되었다. 선행연구에의하면 P.acnes 균에캐모마일에센셜오일을처리하였을때약 8 mm 정도의생육저해환이형성되었다고보고되었으며, 이는본실험의결과와비슷하였다 [17]. 이와같은캐모마일오일의항균력은캐모마일추출물에함유되어있는지용성화합물인 α-bisabolol 및 cyclic ethers 등에의한것으로사료되며 [18], 이에대한추가적인연구는향후연구과제로여겨진다 캐모마일추출물의염증관련유전자발현저해효과 P.acnes 균에의해피부각질형성세포가자극을받으면 TNFα, interleukin-1β (IL-1β), interleukin-6 (IL-6) 및 interleukin-8 (IL-8) 과같은다양한염증성사이토카인이세포외부로분비되어여드름성염증반응이유도된다 [15,19,20]. 염증반응을조절하는주요유전자인 cyclooxygenase-2 (COX-2) 는성장인자 (growth factor), 사이토카인등에의하여발현이유도되 며, 염증반응매개물질인 PGE2 와 NO 의생성에관여함으로써마름버짐, 홍반성낭창과같은염증성피부질환을일으키는것으로알려져있다 [21]. TNF-α 는많은염증반응에관련된중요인자로알려져있으며, 정상적인상태의세포에서는생성되지않으나세포가염증유발인자에의하여자극을받으면분비되는것으로알려져있다 [22]. 또한 IL-1β 는염증초기반응에관여하고있어염증의발생및진행과정에중요한역할을하며, IL-8 은백혈구의탈과립반응을유도함으로써히스타민과같은염증유발인자가분비되도록한다. 분비된염증유발인자는염증부위에부종및통증과같은염증반응을촉진시킨다 [15,22]. 선행연구에의하면피부각질형성세포에 TNF-α 로염증을유도한뒤봉독을처리하였을때, 증가했던 IL-1β 와 IL-8 의분비및발현량이감소한다고보고되었다 [15]. 또한모링가뿌리또는잎추출물을처리하였을때 TNF-α 에의하여유도된 IL-6 의 mrna 발현이농도의존적으로감소한다고보고되었다 [21]. 따라서본연구에서는피부각질형성세포에 TNF-α 를처리하여여드름성염증반응을유도한뒤, 캐모마일추출물을처리하여캐모마일의항염증효과를확인하고자하였다. 그결과, Fig. 3 과같이 TNF-α 의처리에따라 IL-1β, IL-6, IL-8 및 COX-2 유전자의발현이아무것도처리하지않은군과비교하였을때, 각각 1.8, 1.2, 2.0, 1.4 배수증가하였으며, 캐모마일워터처리군에서각각 1.3, 0.3, 1.1, 1.0 배수, 캐모마일오일처리군에서각각 1.1, 0.1, 1.6, 0.4 배수로 TNF-α 단독처리군보다발현량이유의하게감소하였다. 선행연구에의하면캐모마일의항염증효과는페놀성화합물에의한것으로알려져있으며, apigenin, quercetin, patuletin, luteolin 중에서도 luteolin 에의한항염증효과가뛰어난것으로보고되었다 [9]. 결과적으로본실험을통하여캐모마일추출물의항염증효과를확인하였으며, 이와같은효과는 IL-1β, IL-6, IL-8 및 COX-2 과같은염증관련유전자의발현을억제함으로써나타난것임을확인하였다. 또한이러한캐모마일추출물의항염증효과는추출물에함유되어있는 luteolin 과같은페놀성화합물성분에의한것으로예상된다. 4. CONCLUSION 최근화장품시장이확대됨에따라여드름의예방및증상개선을위하여항균및항염효능이있고피부에자극이덜한천연물유래화장료개발에대한연구가많이이루어지고있다 [20]. 따라서본연구에서는캐모마일추출물의항산화효과와세포독성, 항균및항염효과를확인하여천연물유래

14 Korean Society for Biotechnology and Bioengineering Journal 32(1): 9-15 (2017) Fig. 3. Effects of chamomile extracts on IL-1β, IL-6, IL-8 and COX-2 mrna expression levels in HaCaT cells.

6 14 Korean Society for Biotechnology and Bioengineering Journal 32(1): 9-15 (2017) Fig. 3. Effects of chamomile extracts on IL-1β, IL-6, IL-8 and COX-2 mrna expression levels in HaCaT cells. Cells were pre-treated with 10 ng/ml TNF-α for 1 h followed by treatment with chamomile water (C.W) and oil (C.O) for 24 h. The statistical analysis of the data was carried out by use of a t-test. **p<0.01 and ***p<0.001 compared to control. ###p<0.001 was compared to 10 ng/ml TNF-α treated group. N. S.; not significant (each experiment, n=3). 화장료로서캐모마일의가능성에대하여알아보고자하였다. 그결과, 캐모마일워터및오일추출물에서각각 13.3%, 22.75% 의항산화효과가확인되었으며, 캐모마일워터보다캐모마일오일처리군에서더높은항산화효과를확인하였다. 또한 MTT assay 를통하여캐모마일추출물의세포독성검사를실시한결과, 캐모마일추출물처리에따른세포독성은나타나지않았다. 여드름유발균으로알려진 P.acnes 에캐모마일을처리하여항균효과를확인한결과, 캐모마일워터처리군에서는생육저해환이관찰되지않았으며캐모마일오일처리군에서 10±0.0 mm 의생육저해환이형성되었다. 또한 TNF-α 로여드름성염증반응을유도한뒤, 염증관련유전자들의발현변화를통하여캐모마일추출물의항염효과를확인한결과, IL-1β, IL-6, IL-8 및 COX-2 유전자의발현이감소하는것을확인하였다. 결론적으로캐모마일추출물의항산화, 항균및항염효과를확인하였으며, 캐모마일워터보다는캐모마일오일이더뛰어난항여드름효과를지닌것을확인하였다. 따라서, 캐모마일이피부세포에독성이없고항균및항염작용을하는것으로보아여드름증상개선을위한화장료로서이용될가능성이높다고사료된다.

7 HaCaT 세포주에서캐모마일 (Matricaria chamomilla) 추출물의항병원성및항염효과에관한연구 15 Acknowledgements 본연구는 2015 년산학연협력기술개발사업 ( 기업부설연구소신규설치, 과제번호 C ) 에의해수행되었음. REFERANCES 1. Lee, W. G., B. H. Kim, S. W. Jeon, K. S. Kim, H. J. Nam, and Y. B. Kim (2011) The Latest Trends of Treatment for Acne Vulgaris in PubMed. J. Korean Orient. Med. Ophthalmol. Otolaryngol. Dermatol. 24: Jin, B. W., J. H. Ahn, and C. J. Ma (2011) Antibacterial Activity of Some Medicinal Plants against Propionibacterium acnes. Kor. J. Pharmacogn. 42: Yoon, H. J. and D. I. Kim (2015) Review on the Acne Related Articles Published in Korean Medical Journals-Focusing on Experimental Studies Published after J. Korean Obstet. Gynecol. 28: An, B. J., J. T. Lee, C. E. Lee, J. H. Son, J. Y. Lee, and T. S. Park (2005) A Study on the Development of Cosmeceutical Ingredient, Rhododendron mucronulatum, and the Application of Rheology Properties. J. Korean Soc. Appl. Biol. Chem. 48: Eom, J. N. and J. D. Kim (2004) An empirical study on the oriental herbal cosmetics purchase behaviors in women in the Metropolitan area. J. Soc. Cosmet. Scientists Korea. 30: Cho, Y. J., S. J. Yoon, J. H. Kim, and S. S. Chun (2005) Biological Activity of Chamomile (Matricaria chamomilla L.) Extracts. J. Korean Soc. Food Sci. Nutr. 34: Lee, S. H., K. J. Min, K. O. Lee, J. S. Sin, Y. C. Kim (2008) Effect of German Chamomile Oil Application to Atopic Dermatitis Mice on the Change of Serum IgE Level. J. Kor. Soc. Cosm. 14: Srivastava, J. K., E. Shankar, and S. Gupta (2011) Chamomile: A herbal medicine of the past with bright future. Mol. Med. Report. 3: Kim, J. H., S. Y. Kim, Y. N. Hong, Y. S. Kim, and Y. M. Han (2014) Effect of Chamomile Flower Extract on Septic Arthritis due to Candida albicans. Yakhak Hoeji. 58: Abebe, W. (2002) Herbal medication: potential for adverse interactions with analgesic drugs. J. Clin. Pharm. Ther. 27: Weidner, C., S. J. Wowro, M. Rousseau, A. Freiwald, V. Kodelja, H. Abdel-Aziz, O. Kelber, and S. Sauer (2013) Antidiabetic effects of chamomile flowers extract in obese mice through transcriptional stimulation of nutrient sensors of the peroxisome proliferator-activated receptor (PPAR) family. PLoS One. 8: e Ha, J. H., Y. J. Jeong, J. S. Seong, K. M. Kim, A. Y. Kim, M. M. Fu, J. Y. Suh, N. H. Lee, J. Park, and S. N. Park (2015) Antioxidant and Antibacterial Activities of Glycyrrhiza uralensis Fisher (Jecheon, Korea) Extracts Obtained by various Extract Conditions. J. Soc. Cosmet. Sci. Korea. 41: Choi, I. Y., Y. J. Song, and W. H. Lee (2010) DPPH radical scavenging effect and antimicrobial activities of some herbal extracts. Kor. J. Hort. Sci. Technol. 28: Beissert, S., I. Cavazzana, F. Mascia, P. Meroni, S. Pastore, G. Tessari, and G. Girolomoni (2006) Mechanisms of immune-mediated skin diseases: An overview. Clin. Exp. Rheumatol. 24: S Lee, W. R., K. H. Kim, H. J. An, J. Y. Kim, S. M. Han, K. G. Lee, and K. K. Park (2014) The effects of bee venom on tumor necrosis factor (TNF)-α induced inflammatory human HaCaT keratinocytes. Kor. J. Pharmacogn. 45: Sohn, H. Y., Y. S. Kim, E. J. Kum, Y. S. Kwon, and K. H. Son (2006) Screening of anti-acne activity of natural products against Propionibacterium acnes. Kor. J. Microbiol. Biotechnol. 34: Zu, Y. G., H. M. Yu, L. Liang, Y. J. Fu, T. Efferth, X. Liu, and N. Wu (2010) Activities of ten essential oils towards Propionibacterium acnes and PC-3, A-549 and MCF-7 cancer cells. Molecules. 15: Singh, O., Z. Khanam, N. Misra, and M. K. Srivastava (2011) Chamomile (Matricaria chamomilla L.): An overview. Pharmacogn Rev. 5: Taylor, M., M. Gonzalez, and R. Porter (2011) Pathways to inflammation: acne pathophysiology. Eur. J. Dermatol. 21: Choi, J. Y., S. Y. Song, and H. H. Lee (2015) Antibacterial and antiinflammatory activity of corni fructus ethanol extract in Propionibacterium acnes. Kor. J. Aesthet. Cosmetol. 13: Lee, H. J. and Y. C. Chang (2012) Suppression of TNF-α-induced inflammation by extract from different parts of moringa in HaCaT cells. J. Life Sci. 22: Bordon, Y. (2010) Cytokines: Cytokines reach out. Nat. Rev. Immunol. 10:

Journal of Life Science 2011, Vol. 21. No μ μ

Journal of Life Science 2011 Vol. 21. No. 8. 1120~1126 ISSN : 1225-9918 DOI : http://dx.doi.org/10.5352/jls.2011.21.8.1120 μ μ μ α β Journal of Life Science 2011, Vol. 21. No. 8 1121 μ μ 1122 생명과학회지 2011,