ISSN (Print) ISSN (Online) Asian J Beauty Cosmetol 2018; 16(1): R E S E A R C H

Size: px

Start display at page:

Download "ISSN (Print) ISSN (Online) Asian J Beauty Cosmetol 2018; 16(1): R E S E A R C H"

지우 비
5 years ago
Views:

1 ISSN (Print) ISSN (Online) Asian J Beauty Cosmetol 2018; 16(1): R E S E A R C H A R T I C L E Open Access Inhibition Effect of Cnidium officinale Makino Extracts on MMP1 Expression in Human Dermal Fibroblasts Young-Joo Kim Department of Beauty Care and Cosmetics, Osan University, Osan-si, Gyeonggi-do, Korea Corresponding author: Young-Joo Kim, Department of Beauty Care and Cosmetics, Osan University, 45 Chunghak-ro, Osan-si, Gyeonggi-do, 18119, Korea Tel.: Fax: blue@osan.ac.kr Received October 30, 2017 Revised November 14, 2017 Accepted November 22, 2017 Published March 30, 2018 Abstract Purpose: Cnidium officinale Makino (C. officinale) belonged to Umbelliferae family is medical herb native to China and is extensively cultivated in Korea, China and Japan. C. officinale has been used to oriental medicine against pain, inflammation, menstrual disturbance, and anti-vitamin deficiency disease, and also acts as a blood pressure depressant. However, effects of C. officinale are not fully understood in dermis. Therefore, in this study, we identified effects of C. officinale in dermal fibroblast. Methods: Matrix metalloproteinase-1 (MMP1) expression of C. officinale extract was valuated using quantitative real time polymerase chain reaction (qrt-pcr). In addition, transcriptional activity of activator protein 1 (AP-1) and c-jun N-terminal protein kinase (JNK) activity were assayed using luciferase assay and western blot. Results: Extracts of C. officinale decreased MMP1 expression in ultraviolet (UV)-exposed human dermal fibroblasts. Moreover, C. officinale extracts showed decrease of AP-1 transcriptional activity and phospho-jnk in UV-exposed human dermal fibroblasts. Conclusion: Our results suggest that C. officinale extracts decreased MMP1 expression using decreasing AP-1 transcriptional activity and phospho-jnk. Therefore, C. officinale extracts has potential to reduce formation of wrinkle and to use as a cosmetic ingredient. Keywords: MMPs, AP-1, JNK, Cnidium officinale Makino, UV Introduction 피부의기능은내부의기관을외부환경으로부터보호하는것이다 (Kalinin et al., 2002). 피부는표피, 진피, 피하지방으로구분되는데진피는표피밑에있는층으로주로세포외기질 (extracellular matrix proteins, ECM) 로구성되어있다 (Epstein & Munderloh, 1978). UV는피부의구조와기능을조절하는인자중하나로만성적인 UV 노출은피부의광노화를유발한다 (Bae et al., 2010). 이러한노화피부에서는주름, 피부처짐, 색소이상등의특성이나타난다 (Lee et al., 2008; Kim et al., 2005). Matrix metalloproteinases (MMPs) 는구조적으로기질분해효소로써광노화에중요한역할을하고, 세포성장과세포이동, 세포분화, 혈관생성, 세포사멸등에도역시중요하다 (Overall & López-Otin, 2002; Werb, 1997). MMP family중 MMP1은 collagen을분해하여피부내에 collagen과 elastin으로이뤄진원섬유 아교질을분해한다 (Chung et al., 2001). 일반적으로자외선에의해서 MMPs의발현과활성이조절되는데, 자외선을조사받게되면 mitogen-activated protein kinase (MAPK) 에의한신호전달을거쳐전사인자인 AP-1을조절하게된다 (Whitmarsh & Davis, 1996). AP-1은 c-jun과 Fos family 의복합체로 MMP의발현과 collagen 생성저해기능을가진다 (Janulis et al., 1999). 자외선을포함한대부분의외부자극원들은 JNK, ERK, P38과같은 MAPK를조절하고 Jun과 Fos의발현및활성을조절하여세포외기질을분해또는합성한다 (Hibi et al., 1993; Yu et al., 2013). 약용식물인천궁의학명은 Cnidium officinale Makino 으로아시아를중심으로오랜기간동안천연의약품으로사용되어왔다 (Chevallier, 2000). 천궁은다년생식물로산형과에속하고, sedative, anti-anaemia, anti-fungal, smooth muscle relaxing, anti-complementary와같은다양한의학적활성을가지는휘발성의 phthalide 유도체들 Copyright c Korea Institute of Dermatological Sciences. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

2 Inhibition of Cnidium officinale Makino Extracts on MMP1 Expression 을함유하고있으며, 천궁자체역시도혈액순환과염증성질환에탁월한효과가있는것으로알려져있다 (Choi et al., 2002; Jang, 2017; Tahara et al., 1999; Wang et al., 1989). 전통의약에서는 pain, inflammation, menstrual disturbances 등을치료하는효능을가지고있는것으로알려져있고, 울혈과염증억제를통해염증성질환에서혈액순환에의한가려움증을억제할수있다고알려져있다 (Higashi, 1996). 또한 ddy mice와 hepatic carcinoma cells에서항암및항전이활성을가지는것으로밝혀졌다 (Haranaka et al., 1985; Onishi et al., 1998). 하지만천궁추출물이피부에서의어떠한역할을할수있을지에대해서는아직밝혀지지않았다. 그러므로본연구논문에서는천궁의피부세포에서생리학적기능에대해서밝히려고한다. Methods 1. 세포배양및추출물제조인간진피섬유아세포 (normal human dermal fibroblasts, HDFs) 는 Lonza (Basel, Switzerland) 에서구매하였으며, Dulbecco s modified Eagle s medium (DMEM; Gibco, Thermo Fisher Scientific, USA) 에 10% fetal bovine serum (FBS; Sigma-Aldrich, USA) 와 10% penicillin (Gibco, 100 units/ml), 10% streptomycin (Gibco, 100 μ g/ml) 이포함된배지에배양하였다. 인간진피섬유아세포는 15계대이하의세포를사용하여실험을진행하였다. 천궁추출물은천궁 (Nonghyup, Korea) 을잘세척하여서건조하고, 건조된천궁을분쇄기 (SMX-5800LM; Shinil, Korea) 에분쇄하여 100 g을 70% ethanol 1L에넣고 60 에서 30 min 동안 20 khz이상의초음파 (Ultrasonic cleaner 8891; Cole- Parmer, USA) 를이용하여추출하였다. 추출후거름종이 (Whatman No.2; GE Healthcare Life Sciences, USA) 를통해추출하고남은천궁고형파우더를제거하였고, 걸러진추출액은감압농축기 ((EYELA N-3010; Tokyo Rikakikai, Japan) 와동결건조기 (LP 10-30; Ilshin, Korea) 로말려분말화하고, 다시정량후 dimethyl sulfoxide (DMSO; Biopure, Canada) 에녹여실험에사용하였다. 2. Cell viability 측정추출물에의한독성을측정하기위해서 3-(4,5- Dimethylthiazol -2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay 를이용하였다. 96 well plate에인간진피섬유아세포를 cells/well 접종하여 24 h 배양하였다. 24 h 후추출물을 μg/ml 처리하고, 24 h 배양후 0.5 mg/ml의농도가되도록 MTT stock solution (Sigma-Aldrich) 을넣어주었다. 이후 37 에 1 h 배양한후배지를제거하고, 100 μl DMSO 를이용하여 MTT formazan을녹여주었다. 이후 microplate reader (SpectraMax i3x; Molecular Devices, USA) 에서 595 nm의파장으로흡광도를측정하여 cell viability를측정하였다. 3. MMP1 mrna 정량 Total RNA는 Trizol (Invitrogen, USA) 을이용하여추출하였다. 추출된 2 μg total RNA는 oligo dt 2 μg, reverse transcriptase mix (Enzynomics, Korea) 를이용해서 cdna 를합성하였고, 합성된 cdna와 0.2 μg/ml primers (Bionic, Korea; Table 1), 50 mm KCl (Biopure, Canada), 20 mm Tris/HCl ph8.4 (Biopure), 0.8 mm dntp (Takara, Japan), 0.5 U Extaq DNA polymerase (Takara), 3 mm MgCl 2 (Takara), 1X SYBR green (Invitrogen) 를혼합하여 LinegeneK (BioER, China) 을이용하여 qrt-pcr을수행하였다. 측정후 normalization을위해동일 β-actin 발현량에다른 MMP1 발현량을계산하였다. 4. AP-1 promoter 전사활성측정 Reporter plasmid (pgl-tre, 1 μg; Promega, USA) 와 normalization plasmid (pcmv-β-gal, 0.2 μg; Promega) 을 HilyMax (Dojindo, Japan) 를이용하여인간진피섬유아세포에유전자를도입하고, 24 h 동안안정화를시킨후천궁추출물을처리하고, UVA 를조사하였다. 이후수확하여 1X luciferase lysis buffer (Promega, USA) 로세포를용해하고, 4, rpm으로 10 min간원심분리하여 cell lysate 를얻었다. AP-1의전사활성은 cell lysate와 luciferase reagent (Promega) 를혼합하여나오는발광량을 Veritas TM Microplate Luminometer (Turner BioSystems, USA) 로측 Table 1. Sequence of primers Primers MMP1 forward primer MMP1 reverse primer β-actin forward primer β-actin reverse primer Sequence 5 -CTTTCTGGAAGGGCAAGGAC-3 5 -TTGCCTCCCATCATTCTTCA-3 5 -CGACAGGATGCAGAAGGAG-3 5 -ACATCTGCTGGAAGGTGGA

3 인간진피섬유아세포에서천궁추출물의 MMP1 발현조절기전 Figure 1. Cytotoxicity of extracts of C. officinale in human dermal fibroblasts. Effects of C. officinale extracts on the cell viability in human dermal fibroblasts. Cytotoxicity was exhibited as a percentage of control at the indicated concentrations. Values are expressed as M±S.D. from triplicate experiments. C. officinale, Cnidium officinale Makino; M±S.D., mean±standard deviation. 정하였으며, normalization을위해서 β-galatosidase의활성을 O-nitrophenyl-β-D-galactopyranoside (ONPG) assay로측정하여 luciferase assay값을보정하였다. 5. JNK 인산화측정 c-jun N-terminal kinase (JNK) 의활성척도인인산화를보기위해서 JNK와 phospho-jnk의단백질량을 western blot을이용하여측정하였다. 천궁추출물을조건별로처리하고수확된각각의샘플에서 RIPA buffer [50 mm Tris-Cl (ph 7.5; Biopure), 150 mm NaCl (Biopure), 1% NP-40 (Biopure), 0.5% sodium deoxycholate (Biopure), 0.1% sodium dodecyl sulfate (SDS; Biopure), protease inhibitor (Biopure)] 를이용하여세포내모든단백질을추출하였다. 추출된단백질은 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) 에전기영동하여단백질크기별로분리하고, 이를 nitrocellulose membrane (Whatman) 로옮겨 anti-jnk, anti-p-jnk (Cell Signaling Technology, USA) 로 JNK와 phospho- JNK의단백질량을검출하였다. 6. 통계처리본실험의모든실험은 3회반복하여진행하였고, 조건간의차이에대한유의성을입증하는방법으로 Student s t test 을이용하였다. Student s t test 결과 p<0.05의값을유의성있다고판단하고, 유의적인값에는별표 ( * ) 를하였다. Figure 2. Effects of extracts of C. officinale on MMP1 mrna expression in human dermal fibroblasts. Effects of MMP1 mrna expression was measured by qrt-pcr. Expression levels of MMP1 were normalized by β-actin. The graphs are expressed as M±S.D. from triplicate experiments. C. officinale, Cnidium officinale Makino; MMP1, matrix metalloproteinase-1; qrt-pcr, quantitative real time polymerase chain reaction; # p<0.05 compared with control cells; * p <0.05 compared with UVA-exposed cells; UVA, ultraviolet A; M±S.D., mean±standard deviation. Results and Discussion 1. 인간진피섬유아세포에서천궁추출물의세포독성확인인간진피섬유아세포에천궁추출물에의한세포독성을확인하기위해서, 천궁추출물을 μg/ml로인간진피섬유아세포에 24 h 처리후세포생존율을측정하였다. 200 μg/ml 농도까지세포의성장억제가일어나지않는것을확인하였고, 이에따라천궁추출물의독성이없는농도로확인된 200 μg/ml 이하에서 MMP1의발현변화를측정하였다 (Figure 1). 2. 인간진피섬유아세포에서천궁추출물에의한 MMP1 발현변화광노화피부에서는진피탄력도가떨어지는데이는진피섬유아세포에서 MMP1을과도하게발현하여노화가진행되는것으로알려져있다 (Chiang et al., 2012; Cho et al., 2010). 발현된 MMP1은 collagen의분해를일으켜진피탄력을떨어뜨리고이를통해서피부주름이발생하기때문에 (Choi et al., 2016; Lee et al., 2017; Tran et al., 2015) 인간진피섬유아세포에서자외선에의해서증가된 MMP1의발현이천궁추출물에의해서조절되는지확인하였다. qrt- PCR로 MMP1의 mrna 발현량을확인한결과, 자외선을조사하지않은세포에서 MMP1 발현대비 % 까지증가된 MMP1의발현이 100, 200 μg/ml 천궁추출물에의해서각각 , % 까지감소됨을확인할수있었다 (Figure 2)

4 Inhibition of Cnidium officinale Makino Extracts on MMP1 Expression Figure 3. Effects of extracts of C. officinale on AP-1 transcriptional activity in human dermal fibroblasts. Effects of AP-1 transcriptional activity was measured by TREcontained luciferase report plasmid. TRE is DNA binding site of AP-1. AP-1 transcriptional activity was normalized by β-galactosidase assay. The graphs are expressed as the M±S. D. from triplicate experiments. C. officinale, Cnidium officinale Makino; AP-1, activator protein 1; TRE, tet-responsive element; # p <0.05 compared with control cells. * p <0.05 compared with UVA-exposed cells; UVA, ultraviolet A; M±S.D., mean±standard deviation. 3. 인간진피섬유아세포에서천궁추출물에의한 AP-1 전사활성변화광노화에서 MMP1의과발현은주로자외선이 JNK의인산화를증가시키고, 이에따라전사인자인 AP-1의전사활성이증가되어 MMP1의발현을높이는것으로알려져있다 (Hwang et al., 2013; Qin et al., 2014). 때문에자외선조사환경에서 MMP1의발현을조절하는것으로알려져있는 AP-1의전사활성변화를확인하였다. Figure 3에서보듯이, 자외선에의해 % 증가된 AP-1 전사활성이 100, 200 μg/ml 천궁추출물에의해서 , % 로농도의존적으로감소함을확인할수있었다. 4. 인간진피섬유아세포에서천궁추출물에의한 JNK 활성변화자외선에의한광노화에서 AP-1의조절은주로 JNK에의해서조절되기때문에 JNK의활성을확인하였다 (Hwang et al., 2013; Qin et al., 2014). JNK는인산화를통해서활성이조절되기때문에 phosphor-jnk의양을비교하여 JNK 활성을확인하였다. Figure 4에서보는바와같이자외선에의해서급격하게증가된 phosphor-jnk가천궁추출물의처리에의해서농도의존적으로감소함을확인할수있었다. Figure 4. Effects of extracts of C. officinale on JNK phosphorylation in human dermal fibroblasts. Effects of JNK activity was measured by western blot of JNK and phospho-jnk. After western blot, each band density was digitized by densitometer. JNK activity was normalized by normal JNK. The graphs are expressed as the M±S.D. from triplicate experiments. C. officinale, Cnidium officinale Makino; JNK, c-jun N-terminal protein kinase; # p <0.05 compared with control cells; * p<0.05 compared with UVAexposed cells; UVA, ultraviolet A; M±S.D., mean±standard deviation. Conclusion 본논문은천궁추출물의항노화기전을확인하고자하였다. 본논문의결과에서보는것과같이천궁추출물은 200 μg/ml 이하의농도에서는세포독성이없는것을확인하였으며, 따라서 200 μg/ml이하에서자외선에의해증가된 MMP1발현을감소시키는것을확인하였다. 또한이러한 MMP1의조절은 mitogen-activated protein kinase (MAPK) 의하나인 JNK 활성변화를통한 AP-1의전사활성을감소시켰기때문으로확인할수있었다. 자외선은세포내활성산소를증가시키고이를통해서 JNK를활성화시킨다. AP-1의전사활성이증가하고, AP-1의조절을받는 MMP1 의발현이증가하여피부노화를유도하는것으로알려져있다. 천궁추출물은많은논문을통해서다양한항산화물질이함유되어있는것으로알려져있다. 이러한항산화물질이작용하여자외선에의한활성 134

5 인간진피섬유아세포에서천궁추출물의 MMP1 발현조절기전 산소생성을억제함으로써 JNK의인산화를억제할수있을것으로예측된다. 본연구결과를통해서천궁추출물은피부의 collagen 을분해하는 MMP1의발현을감소시켜 collagen 의분해를조절할수있는화장품원료임을확인하였고, 때문에새로운주름개선기능성화장품성분으로서가능성을제시해주고있다. Acknowledgements 이논문은 2017년도오산대학교교내학술비지원에의해서수행됨. References Bae JY, Choi JS, Kang SW, Lee YJ, Park J, Kang YH. Dietary compound ellagic acid alleviates skin wrinkle and inflammation induced by UV-B irradiation. Experimental Dermatology, 19: , Chiang HM, Chen HC, Lin TJ, Shih IC, Wen KC. Michelia alba extract attenuates UVB-induced expression of matrix metalloproteinases via MAP kinase pathway in human dermal fibroblasts. Food and Chemical Toxicology, 50: , Chung JH, Seo JY, Choi HR, Lee MK, Youn CS, Rhie G, Cho KH, Kim KH, Park KC, Eun HC. Modulation of skin collagen metabolism in aged and photoaged human skin in vivo. Journal of Investigative Dermatology, 117: , Cho S, Lee DH, Won CH, Kim SM, Lee S, Lee MJ, Chung JH. Differential effects of low-dose and high-dose betacarotene supplementation on the signs of photoaging and type I procollagen gene expression in human skin in vivo. Dermatology, 221: , Choi HS, Kim MSL, Sawamura M. Constituents of the essential oil of cnidium officinale Makino, a Korean medicinal plant. Flavour and Fragrance Journal, 17: 49-53, Choi SJ, Lee SN, Kim K, Joo da H, Shin S, Lee J, Lee HK, Kim J, Kwon SB, Kim MJ, et al. Biological effects of rutin on skin aging. International Journal of Molecular Medicine, 38: , Chevallier A. The encyclopedia of medicinal plants. Dorling Kindersley Publishing, London, pp , Epstein EH Jr, Munderloh NH. Human skin collagen. presence of type I and type III at all levels of the dermis. The Journal of Biological Chemistry, 253: , Haranaka K, Satomi N, Sakurai A, Haranaka R, Okada N, Kobayashi M. Antitumor activities and tumor necrosis factor producibility of traditional Chinese medicines and crude drugs. Cancer Immunology, Immunotherapy, 20: 1-5, Hibi M, Lin A, Smeal T, Minden A, Karin M. Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-jun activation domain. Genes & Development, 7: , Higashi K. The therapeutic effect of Unsei-in on facial redness (inflammatory congestion) in atopic dermatitis. The Japanese Journal of Oriental Medicine, 46: , Hwang BM, Noh EM, Kim JS, Kim JM, You YO, Hwang JK, Kwon KB, Lee YR. Curcumin inhibits UVB-induced matrix metalloproteinase-1/3 expression by suppressing the MAPK-p38/JNK pathways in human dermal fibroblasts. Experimental Dermatology, 22: , Jang YA. Efficacy of a cosmetic material from complex extracts of Vaccinium spp., Phellinus linteus, Castanea crenata, and Cimicifuga heracleifolia. Asian Journal of Beauty and Cosmetology, 15: , Janulis M, Silberman S, Ambegaokar A, Gutkind JS, Schultz RM. Role of mitogen-activated protein kinases and c-jun/ap-1 trans-activating activity in the regulation of protease mrnas and the malignant phenotype in NIH 3T3 fibroblasts. The Journal of Biological Chemistry, 274: , Kalinin AE, Kajava AV, Steinert PM. Epithelial barrier function: assembly and structural features of the cornified cell envelope. BioEssays, 24: , Kim HH, Shin CM, Park CH, Kim KH, Cho KH, Eun HC, Chung JH. Eicosapentaenoic acid inhibits UV-induced MMP- 1 expression in human dermal fibroblasts. Journal of Lipid Research, 46: , Lee CW, Choi HJ, Kim HS, Kim DH, Chang IS, Moon HT, Lee SY, Oh WK, Woo ER. Biflavonoids isolated from Selaginella tamariscina regulate the expression of matrix metalloproteinase in human skin fibroblasts. Bioorganic & Medicinal Chemistry, 16: , 135

6 Inhibition of Cnidium officinale Makino Extracts on MMP1 Expression Lee JJ, Kim KB, Heo J, Cho DH, Kim HS, Han SH, Ahn KJ, An IS, An S, Bae S. Protective effect of Arthrospira platensis extracts against ultraviolet B-induced cellular senescence through inhibition of DNA damage and matrix metalloproteinase-1 expression in human dermal fibroblasts. Journal of Photochemistry and Photobiology B: Biology, 173: , Overall CM, López-Otin C. Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nature Reviews Cancer, 2: , Onishi Y, Yamaura T, Tauchi K, Sakamoto T, Tsukada K, Nunome S, Komatsu Y, Saiki I. Expression of the antimetastatic effect induced by Juzen-taiho-to is based on the content of Shimotsu-to constituents. Biological and Pharmaceutical Bulletin, 21: , Qin Z, Voorhees JJ, Fisher GJ, Quan T. Age-associated reduction of cellular spreading/mechanical force upregulates matrix metalloproteinase-1 expression and collagen fibril fragmentation via c-jun/ap-1 in human dermal fibroblasts. Aging Cell, 13: , Tahara E, Satoh T, Toriizuka K, Nagai H, Nunome S, Shimada Y, Itoh T, Terasawa K, Saiki I. Effect of Shimotsu-to (a Kampo medicine, Si-Wu-Tang) and its constituents on triphasic skin reaction in passively sensitized mice. Journal of Ethnopharmacology, 68: , Tran TA, Ho MT, Song YW, Cho M, Cho SK. Camphor induces proliferative and anti-senescence activities in human primary dermal fibroblasts and inhibits UVinduced wrinkle formation in mouse skin. Phytotherapy Research, 29: , Wang JD, Narui T, Kurata H, Takeuchi K, Hashimoto T, Okuyama T. Hematological studies on naturally occurring substances. II. effects of animal crude drugs on blood coagulation and fibrinolysis systems. Chemical and Pharmaceutical Bulletin, 37: , Werb Z. ECM and cell surface proteolysis: regulating cellular ecology. Cell, 91: , Whitmarsh AJ, Davis RJ. Transcription factor AP-1 regulation by mitogen-activated protein kinase signal transduction pathways. Journal of Molecular Medicine, 74: , Yu H, Huang J, Wang S, Zhao G, Jiao X, Zhu L. Overexpression of Smad7 suppressed ROS/MMP9-dependent collagen synthesis through regulation of heme oxygenase-1. Molecular Biology Reports, 40: ,

7 인간진피섬유아세포에서천궁추출물의 MMP1 발현조절기전 국문초록 인간진피섬유아세포에서천궁추출물의 MMP1 발현조절기전 김영주오산대학교뷰티케어코스메틱과, 경기도오산시, 한국 목적 : 천궁은산형과에속하는약초로써중국이원산지이고, 한국, 중국, 일본에서많이사용되어왔다. 천궁은진통, 항염증, 월경개선, 비타민결핍증억제및고혈압에약으로사용되어왔다. 하지만아직피부에서의역할은알려지지않았다. 때문에본연구에서는천궁의인간진피섬유아세포에서의역할을확인하고자한다. 방법 : 천궁에의한 matrix metalloproteinase-1 (MMP1) 의발현변화는 quantitative real time polymerase chain reaction (qrt-pcr) 로확인하였고, 추가로 activator protein 1 (AP-1) 과 c-jun N-terminal protein kinase (JNK) 의활성은각각 luciferase assay 와 western blot 을통해서확인하였다. 결과 : 자외선에의해서증가된 MMP1의발현이천궁추출물에의해다시감소되었다. 또한자외선이조사된인간진피섬유아세포에서천궁추출물은 AP-1의전사활성을감소시키고, JNK의인산화를감소시키는것으로확인되었다. 결론 : 본논문의연구결과를통해서천궁추출물은 AP-1 과 JNK 인산화를조절함으로써 MMP1의발현을감소시킴을알수있다. 때문에천궁추출물은피부의 collagen 분해하는 MMP1의발현을감소시켜 collagen 의분해를조절할수있는화장품원료로서가능성을가지고있다. 핵심어 : MMPs, AP-1, JNK, Cnidium officinale Makino, UV 이논문은 2017 년도오산대학교교내학술비지원에의해서수행됨. 참고문헌 장영아. 블루베리, 상황, 율피, 승마복합추출물의화장품소재로서의효능. 아시안뷰티화장품학술지, 15: ,

8 Inhibition of Cnidium officinale Makino Extracts on MMP1 Expression 中文摘要川芎提取物对人真皮成纤维细胞 MMP1 表达的抑制作用金英珠乌山大学美容化妆品学科, 京畿道乌山市, 韩国目的 : 川芎属于伞形科, 是中国本土的药用植物, 在韩国中国和日本广泛种植川芎已被用于治疗疼痛, 炎症, 月经紊乱和抗维生素缺乏症的东方药物, 也可用作降压药然而, 在皮肤中并不完全了解川芎的效果因此, 在本研究中, 确认川芎在人真皮成纤维细胞中的作用方法 : 为确认川芎的作用, 利用 quantitative real time polymerase chain reaction (qrt-pcr) 测量 matrix metalloproteinase-1 (MMP1) 的表达, 利用 luciferase assay 和 western blot 分别测量 activator protein 1 (AP-1) 和 c-jun N-terminal protein kinase (JNK) 的活性结果 : 川芎提取物降低紫外线照射增加的 MMP1 的表达此外, 川芎提取物在人真皮成纤维细胞中, 降低 AP-1 的转录活性和 JNK 的磷酸化结论 : 研究结果表明川芎提取物通过调节 AP-1 和 JNK 的磷酸化降低 MMP1 的表达因此, 川芎提取物减少了分解胶原蛋白质的 MMP1 的表达, 作为调节胶原蛋白质分解的化妆品原料充分具有可行性关键词 : MMPs,AP-1,JNK, 川芎, 紫外线 138

139~144 ¿À°ø¾àÄ§

139~144 ¿À°ø¾àÄ§ 2 139 DOI : 10.3831/KPI.2010.13.2.139 2 Received : 10. 04. 08 Revised : 10. 04. 26 Two Case Report on Wrist Ganglion Treated with Scolopendrid Pharmacopuncture Accepted : 10. 05. 04 Key Words: Wrist Ganglion,