Journal of Nutrition and Health (J Nutr Health) 2016; 49(5): 269 ~ 276 http://dx.doi.org/10.4163/jnh.2016.49.5.269 pissn 2288-3886 / eissn 2288-3959 Research Article 자외선조사와병행된녹차추출물식이공급이무모생쥐의표피보습개선및유리아미노산생성관련대사에미치는영향 * 최수민 신지혜 이보민 조윤희 경희대학교동서의학대학원의학영양학과 Dietary effect of green tea extract on hydration improvement and metabolism of free amino acid generation in epidermis of UV-irradiated hairless mice* Choi, Sumin Shin, Jihye Lee, Bomin Cho, Yunhi Department of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University, Yongin 17104, Korea ABSTRACT Purpose: Ultraviolet (UV) irradiation decreases epidermal hydration, which is maintained by reduction of natural moisturizing factors (NMFs). Among various NMFs, free amino acids (AA) are major constituents generated by filaggrin degradation. This experiment was conducted to determine whether or not dietary supplementation of green tea extract (GTE) in UV-irradiated mice can improve epidermal levels of hydration, filaggrin, free AAs, and peptidylarginine deiminase-3 (PAD3) expression (an enzyme involved in filaggrin degradation). Methods: Hairless mice were fed a diet of 1% GTE for 10 weeks in parallel with UV irradiation (group UV+1%GTE). As controls, hairless mice were fed a control diet in parallel with (group UV+) or without (group UV-) UV irradiation. Results: In group UV+, epidermal levels of hydration and filaggrin were lower than those in group UV-; these levels increased in group UV+1% GTE to levels similar to group UV-. Epidermal levels of PAD3 and major AAs of NMF, alanine, glycine and serine were similar in groups UV- and UV+, whereas these levels highly increased in group UV+1% GTE. Conclusion: Dietary GTE improves epidermal hydration by filaggrin generation and degradation into AAs. KEY WORDS: green tea extract (GTE), filaggrin, peptidylarginine deiminase-3 (PAD3), natural moisturizing factor (NMF) 서론 피부의최외곽층인표피는인체내수분의손실을막고보습을유지하여피부에유연성을부여하고, 피부장벽역할을수행한다. 1,2 표피의각질층은세포사이의다중층상구조를이루어체액손실을억제하는피지막과각질세포내의친수성성분으로수분과결합하여흡습성에직접적인영향을미치는자연보습인자 (natural moisturizing factor, NMF) 에의해피부의보습을유지한다. 자연보습인자에는유리아미노산, pyrrolidone carboxylic acid (PCA), lactate, urea 등이있으며그중유리아미노산은약 40% 의높은함량을차지한다. 1,3 유리아미노산은 filaggrin의대사를거쳐생성되는데과립층의 keratohyalin granule 안에 있는 profilaggrin은불활성및불용성인상태로저장되어있다가탈인산화및단백분해과정을거쳐 filaggrin으로분해되어각질층으로방출된다. 4 Filaggrin은세포골격을형성하는 keratin filament와결합하여 matrix protein으로피부장벽을구성하며 peptidylarginine-deiminase3 (PAD3) 등의효소에의해유리아미노산으로최종분해되어자연보습인자의역할을수행한다. 5 외부적인스트레스중하나인자외선은피부건조및과증식, 주름형성과같은광노화뿐만아니라면역억제와피부세포의 DNA 손상을일으키고 6-8 표피 filaggrin의발현및유리아미노산으로분해되는과정의변화를야기하는것으로알려져있다. 1,9 광노화에따른피부건조, 주름형성등을완화시키기위해여러기능성식이소재개발이이루어지고 Received: June 10, 2016 / Revised: July 6, 2016 / Accepted: September 5, 2016 *This study was supported by a grant of the Korean Health Technology R & D Project, Ministry of Health & Welfare, Republic of Korea (Grant No. HN13C0076). To whom correspondence should be addressed. tel: +82-31-201-3817, e-mail: choyunhi@khu.ac.kr 2016 The Korean Nutrition Society This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
270 / 녹차추출물섭취에의한피부보습개선효능 있는데 10,11 이중녹차의카테킨은피부의항염증, 항암및항산화효과를입증한연구가많이보여졌으며, 12,13 자외선에의해유도되는종양형성과광노화를저해하는물질로써 14,15 치료및화장품으로상용되고있다. 선행연구를통해녹차의카테킨중주요활성물질인 epigallocatechin- 3-gallate (EGCG) 로처리한각질형성세포에서 filaggrin의발현을유도함이보고되었다. 16 그러나녹차추출물식이공급을통한피부보습관련 filaggrin 대사기전및유리아미노산에대한연구는미흡한실정이다. 이에본연구에서는무모생쥐에자외선조사와함께녹차추출물을식이공급하고피부보습의변화를비롯하여, 표피 filaggrin과유리아미노산의함량및 filaggrin 분해관련효소의단백질발현에미치는영향을살펴보았다. 연구방법 실험동물생후 6주된암컷무모생쥐 [Albino hairless mouse (SKH- 1)] 를 ( 주 ) 오리엔트바이오에서공급받아실험식이로사육하기전 1주간고형배합사료로적응시켰다. 체중에따른난괴법을이용하여자외선을조사하지않은정상대조군 (UV 군, n = 5), 자외선조사군 (UV+ 군, n = 5) 및자외선조사와함께녹차추출물 (green tea extract, GTE) 을전체식이의 1% 수준에서공급한실험군 (UV+1%GTE군, n = 5) 으로나누었다. 사육기간동안사료와물을마음껏섭취하도록하였으며온도는 24 ± 2 o C, 습도는 55 ± 10% 를유지하고광주기와암주기가매일 12시간이되도록조절하였다. 본동물실험에대한프로토콜은경기과학기술진흥원의동물실험윤리위원회에의해승인되었다 ( 승인번호 : 2013-12-0006). 자외선조사사료투여개시일로부터 1주일에 3회씩 UVB lamp (30% 의 UVA를함유함 : TL 20W/12RS SLV, wave length 290~ 390 nm, Phillips, Netherlands) 로자외선조사를하였다. 자외선조사량의측정은 UV meter (VARIOCONTROL, Waldmann ver. 2.03, Germany) 를사용하였다. 13 1주차에는 1MED (minimal erythema dose; 75 mj/cm 2, 15 min) 로시작해서 2주차에 2MED (150 mj/cm 2, 30 min), 3주차에 3MED (225 mj/cm 2, 45 min), 4주차부터 10주차까지 4MED (250 mj/cm 2, 50 min) 를동물의등부위 ( 등중앙기준, 2 cm 3 cm) 에조사하였다. 17 실험식이 UV- 군과 UV+ 군은 GTE 가첨가되지않은일반식이 (AIN-93G) 를공급하였다. 13 UV+1%GTE군은전체식이무게의 1.0% 에상응하는 GTE [Catechin 50%, EGCG 28% 함유 ] 를식이에첨가하여공급하였다. 식이 1% 의 GTE 첨가수준은독성을유발하지않으며 18 피부산도개선의효과가나타난예비실험을바탕으로결정하였다. 19 GTE 시료첨가에따른건분식이무게차이는 corn starch 를감하여조정하였으며, GTE 시료는 ( 주 ) 아모레퍼시픽 (South Korea) 에서공급받았다. 각실험군의식이조성은 Table 1과같다. 피부보습측정피부의보습측정은 Corneometer CM825 (Courage & Khazaka, Cologne, Germany) 를사용하여 10주차에측정하였다. 측정시각개체별측정부위는동일하게등중앙부분으로하였으며측정면에대한압력을일정하게눌러총 3회측정하여수치의평균값을기록하였다. 동물의희생과표피분리 10주간의실험식이및자외선조사후경추탈골로희생시키고등부위의피부조직을얻었다. 피부조직은 Dispase II (2.4 unit/ml, Roche, Germany) 와 Hank solution (Hank s balanced salt solution) 이혼합된용액 (1:1 v/v) 에담가 4 o C 에서 16시간동안처리한후표피를분리하였다. 11 표피 filaggrin 추출및분석분리한표피는 2 mm EDTA, 9 M urea, 2% SDS 및 protease inhibitor cocktail 가포함된 50 mm Tris buffer (P- Table 1. Diet composition and UV irradiation of experimental groups (g/kg) Ingredients Experimental group 1) UV- UV+ GTE 2) Corn starch 372 372 362 Sucrose 200 200 200 Cellulose 50 50 50 Corn oil 100 100 100 Vitamin mix 3) 10 10 10 Mineral mix 4) 35 35 35 L-cystine 3 3 3 Casein 5) 230 230 230 GTE - - 10 UV irradiation - + + 1) Groups UV- and UV+, hairless mice fed a control diet without (group UV-) or with (group UV+) UV irradiation for 10 weeks; Group GTE, hairless mice fed a diet containing 1.0% green tea extract (GTE) in parallel with UV irradiation for 10 weeks. 2) GTE: green tea extract 3) AIN-93 vitamin mix #310025 (Dyets Inc., Bethlehem, PA) 4) AIN-93G salt mix #210025 (Dyets Inc., Bethlehem, PA) 5) Casein (nitrogen 6.25), 870 g/kg
Journal of Nutrition and Health (J Nutr Health) 2016; 49(5): 269 ~ 276 / 271 8340, Sigma Chemical Co., St. Louis, MO, USA) 를처리하여 20 polytron으로분쇄하였다. 분쇄한조직을 12,000 g 에서 4 o C로 20분동안원심분리후상층액을얻었고, 단백질정량을하였다. 동량의단백질 (20 µg/well) 을 4% (profilaggrin 발현 ) 또는 8% (filaggrin 발현 ) SDS/PAGE gel에전기영동후 nitrocellulose membrane에흡착시켰다. Membrane을 1차항체인 anti-rabbit polyclonal filaggrin antibody (Enzo Biochem Inc., NY, USA, cat no. p20930) 를 1/1,000의농도로희석한 PBS solution과함께 4 o C에서 17시간동안반응시키고 PBST solution으로 3회세척하였다. 이어서 anti-rabbit IgG-HRP (Bethly Laboratories Inc., Mongomery, TX, USA, cat no. A120-101P) 로 2시간동안실온에서반응시킨후 PBST solution으로 3회세척하고 Super Signal West Pico Chemiluminescent Substrate (Thermo Scientific, Rockford, IL, USA) 를통해 detection 하였다. 각 band는 imaging densitometer (Lab work version 4.6, UVP, upland, CA, USA) 를이용하여측정한후 actin의발현을기준으로보정하였다. 표피 PAD3 발현분석분리된표피에 0.5% Nonidet P-40 (TE-NP40 buffer extract), 10 mm EDTA, 40 mm Tris-HCl (ph 7.5), protease inhibitor 및 Tris-EDTA buffer을첨가하여 polytron을사용하여분쇄하였다. 5 분쇄한조직은원심분리 (1,2000 g, 4 o C, 20 min) 하여상층액을얻었고, 단백질정량후 8% SDS/PAGE gel에시료 20 µg/well을전기영동하여 nitrocellulose membrane에흡착시켰다. Membrane은 1차항체 anti- goat polyclonal PAD3 antibody (cat no. sc-55693, Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) 및 2차항체 anti-goat IgG-HRP (bethly Laboratories Inc., Montgomery, TX, USA) 를이용하여동일한방법으로반응시킨뒤 Super Signal West Pico Chemiluminescent substrate를통해 detection하여발색하였다. 각 band의 intensity 는 imaging densitometer 를사용하여측정후 actin 의발현을기준으로정량되었다. : 45 : 10, volume) 의용액을 Model 510 solvent delivery system Pico Tag column (3.9 300 nm, 4 µm) 에서유속 1.0 ml/min의속도로아미노산을분리하고각 2487 UV detector (254 nm, Waters) 를사용하여함량분석후사용표피무게의정량값으로보정하였다. 통계분석실험결과는 SPSS statistics 22 program (statistical package for social science) 을사용하여통계처리하였다. 각실험군별결과는평균 (mean) 과표준편차 (standard deviation, SD) 로나타냈으며, 각군간비교는 one-way ANOVA 로분석한후 Tukey s multiple range test로 p < 0.05 수준에서검증하였다. 결 피부보습측정자외선조사및녹차추출물식이공급후 10주차에피부보습측정결과는 Fig. 1에나타내었다. 자외선조사 10주에측정한보습함량은자외선조사군인 UV+ 군이정상대조군인 UV-군보다유의적으로낮았는데이결과는지속적인자외선조사에의해피부보습함량이감소하였음을의미한다. 반면에자외선조사와병행하여녹차추출물을 10 주간공급한실험군인 UV+1%GTE 군의보습은 UV+ 군에비해유의적으로높았으나 UV-군의함량에는미치지못하였다. 이는전체식이 1.0% 에달하는녹차추출물의식이공급이자외선조사에의해유도된피부건조를완화시켰음을의미한다. 과 표피유리아미노산함량분석표피조직에 15% trichloroacetic acid (TCA) 용액을첨가하여 polytron을이용해분쇄하고 4 o C에서 15시간동안반응시켜단백질을침전시켰다. 12,000 g에서 10분간원심분리를하여 5 상층액을분리한후 0.45 µm filter (Millipore, USA) 를사용해상층액을여과하여 high performance liquid chromatography (HPLC) 를통해각유리아미노산의농도를분석하였다. Acetonitrile : methanol : water (45 Fig. 1. Altered epidermal hydration of groups. Hairless mice fed a control diet without UV irradiation for 10 weeks (group UV-); UVirradiated hairless mice fed a control diet (group UV+) or a diet supplemented with 1.0% green tea extract (group UV+1%GTE) for 10 weeks. Values are mean ± SD (n = 5). Values with different alphabetical letters are significantly different (p < 0.05) using oneway ANOVA and Tukey's multiple range test.
272 / 녹차추출물섭취에의한피부보습개선효능 표피 profilaggrin 및 filaggrin의함량변화표피의 profilaggrin 및 filaggrin 함량변화를 western blot assay로분석하였고그결과를 Fig. 2에나타내었다. 자연보습인자의전구체인 filaggrin은각질형성세포가분화단계에서발현시키는단백질중하나로피부의보습및피부막기능의지표로사용된다. 4,21 UV+ 군의 profilaggrin 함량은정상대조군인 UV-군과유의적인차이가없었으나, filaggrin 함량은 UV-군에비해유의적으로낮았다. 반면 UV+1%GTE 군의 profilaggrin 함량은 UV-군및 UV+ 군보다현저히높았다. Filaggrin의함량은 UV+ 군에비해유의적으로높았으며 UV-군의함량과유사하였다. 이는녹차추출물의식이공급이 pro-filaggrin의합성을촉진시키고자외선에의한표피 filaggrin 함량의감소를정상화시켰음을의미한다. PAD3 단백질발현변화 PAD3는 filaggrin의분해과정에서 filaggrin의 deimination 을통해유리아미노산의분해를유도한다. 22 표피 PAD3의 단백질발현을 western blot assay로분석한결과 (Fig. 3) UV+ 군의 PAD3 단백질발현은 UV-군에비해유의적으로낮았다. 이는자외선조사가 PAD3의발현을억제시켰음을의미한다. UV+1%GTE군에서는 UV+ 군에비해유의적으로높았으며 UV-군의함량보다유의적으로높았다. 따라서, 녹차추출물의식이공급이 PAD3 발현을현저하게활성화함을의미한다. 표피유리아미노산함량변화 Filaggrin은 serine, glycine, glutamate, histidine, alanine 등의아미노산을다량함유한단백질로 23 분해과정에서유리아미노산으로전환되어아미노산의친수성성질에의해자연보습인자로서표피의보습유지를담당한다. 3,24 유리아미노산분석결과 serine 과 glycine, alanine 은 UV- 군과 UV+ 군에서유의적인차이를보이지않은반면, UV+1%GTE군에서는두군에비해유의적으로높았다. Histidine, arginine, asparagine 또한 UV- 군과 UV+ 군에서유의적인차이는나타나지않았으나, UV+1%GTE군에서는 UV-군에비해유의적으로높았다. UV+ 군에서 glutamine 및 tryptophan은 UV+1%GTE군과유사하였다. 그외 Fig. 2. Altered protein levels of profilaggrin and filaggrin in the epidermis of groups. Hairless mice fed a control diet without UV irradiation for 10 weeks (group UV-): UV-irradiated hairless mice fed a control diet (group UV+) or a diet supplemented with 1.0% green tea extract (group UV+1%GTE) for 10 weeks (A) Representative expression of profilaggrin and filaggrin proteins in the epidermis of groups (B) Signal intensities from multiple experiments of (A) were quantified and the integrated areas were normalized, first to the corresponding value of actin and then to the signal observed in the normal control group (group UV-). Values are mean ± SD (n = 5). Values with different alphabetical letters in profilaggrin and filaggrin are significantly different (p < 0.05) using one-way ANOVA and Tukey's multiple range test. Fig. 3 Altered protein level of peptidylarginine deiminase-3 in the epidermis of groups. Hairless mice fed a control diet without UV irradiation for 10 weeks (group UV-); UV-irradiated hairless mice fed a control diet (group UV+) or a diet supplemented with 1.0% green tea extract (group UV+1%GTE) for 10 weeks (A) Representative expression of peptidylarginine deiminase-3 (PAD3) protein in the epidermis of groups (B) Signal intensities from multiple experiments of (A) were quantified and the integrated areas were normalized, first to the corresponding value of actin and then to the signal observed in the normal control group (group UV-). Values are mean ± SD (n = 5). Values with different alphabetical letters in PAD3 is significantly different (p < 0.05) using one-way ANOVA and Tukey's multiple range test.
Journal of Nutrition and Health (J Nutr Health) 2016; 49(5): 269 ~ 276 / 273 Fig. 4. Free amino acid contents in the epidermis of groups. Hairless mice fed a control diet without UV irradiation for 10 weeks (group UV-); UV-irradiated hairless mice fed a control diet (group UV+) or a diet supplemented with 1.0% green tea extract (group UV+1%GTE) for 10 weeks. Data are mean ± SD (n = 5). Values with different alphabetical letters in each amino acids are significantly different (p < 0.05) using one-way ANOVA and Tukey s multiple range test. isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine, aspartate, glutamate, proline 및 tyrosine 의함량은모든군에서군간유의성이없었다. 이는녹차추출물의식이공급이자연보습인자인 serine, glycine, arginine 등보습인자역할을하는주요유리아미노산함량증가시켜피부보습을증가시켰음을의미한다. 고 지속적인자외선조사에의해초래되는광노화는내인성노화와달리산화스트레스증가와함께피부보습및탄력감소에의한주름을야기하고, 색소침착, 피부암등을유발하는것으로보고되어진다. 7,8 주 3회 15~50분간매우높음수준의자외선지수 3.3배에해당하는양을 10주동안조사하였을때, 피부보습함량이낮았는데이결과는사전연구에서자외선비조사군에비해자외선조사군의피부의보습함량이낮고경피수분손실 (transepidermal water loss, TEWL) 이높은결과와일치하였다. 25 반면자외선조사를병행한녹차추출물의식이공급은 UV+ 군에비해피부보습함량이높았다. 즉, 녹차추출물의식이공급은지속적인자외선조사로야기된피부건조를완화시켰음을의미한다. 사전연구에서 filaggrin을구성하는아미노산과표피내자연보습인자로작용하는유리아미노산의구성이유사하게나타남에의해자연보습인자로작용하는표피유리아미노산의유일한전구체는 filaggrin임이입증되었다. 23,26 과립층의 keratohyalin granule 내에존재하는 profilaggrin 은표피의칼슘기울기가증가함에따라 phosphatase 찰 (PPase), 27 furin 28 등의효소에의해탈인산화와단백분해과정을거쳐 histidine 이풍부한 filaggrin 으로분해된다. 4,26 자외선을조사한 UV+ 군에서 profilaggrin의함량은 UV-군과의차이가나타나지않았으나, filaggrin의함량은 UV-군에비해낮게발현되었다. 이는자외선조사에의해피부장벽이손상되면표피의칼슘기울기가감소하고경피수분손실이증가한다는기존연구와함께 29 지속적인자외선조사가 profilaggrin이 filaggrin으로분해되는과정에장애를일으킨것을의미한다. 반면, UV+1%GTE군의 profilaggrin 함량은 UV+ 군에비해높았고, filaggrin의함량은 UV-군과유사한수준으로높았다. 사전연구에서녹차의주요카테킨인 EGCG가 profilaggrin발현의 regulator 4,30 로작용하는 AP-1 (activator protein-1) 을활성화하며, 31 EGCG에대한 in vitro 실험을통해정상각화세포에서 filaggrin 발현의증가가보고되었다. 16 즉, 녹차추출물의식이공급이표피 profilaggrin 함량을높이고, 자외선에의한대사장애를정상화시켰음을나타낸다. Profilaggrin은 filaggrin으로분해후과립층과각질층경계부에서 keratin intermediate filaments와결합하여세포의골격을이루는거대원섬유를형성한다. 4,21 Keratin과결합된 filaggrin은 PADs에의해아미노산의잔기가 arginine에서 citrulline으로치환되는 deimination 과정을거쳐 keratin 과분리되고 caspase-14, 32 bleomycin hydrolase4 등의효소에의한단백분해과정을거쳐유리아미노산으로분해된다. Filaggrin의 deimination 에는칼슘의존형효소인 PAD1과 PAD3가관여하는데, 그중 PAD3의발현장소는모발과표피로제한되며, 사전연구에서 PAD3의발현은과립층및각질층에서 (pro)filaggrin과함께발현되었다. 5,22
274 / 녹차추출물섭취에의한피부보습개선효능 PAD3는 Profilaggrin이 filaggrin으로분해된후유리아미노산으로분해되는과정의도입부로써중요한역할을하며, filaggrin의 citrullination에 PAD3가관여되지못하면표피의항상성을손상시키고피부의장벽기능을잃게한다. 33 PAD3의발현은 UV+1%GTE군이 UV-군및 UV+ 군보다현저히높았다. EGCG를처리한각질형성세포에서 filaggrin을유리아미노산으로분해하는 caspase-14효소의발현이증가되었다는선행연구도보고되었는데, 34 이를종합하여녹차추출물의식이공급이 PAD3 및 filaggrin의분해에관여하는효소의발현을촉진하여궁극적으로유리아미노산의생성이높게나타난것으로사료되며, 추후 PAD3이외 caspase-14 등아미노산으로분해되는과정에관여하는효소에대해서도추가적인연구가필요할것으로여겨진다. 자연보습인자인유리아미노산중 serine과 glycine은총유리아미노산의 40% 을차지하며 alanine 또한약 10% 가량함유되어있다고보고되었다. 또한, 표피의건조화및과증식에의해 serine과 glycine의급격한농도감소가나타남에따라 35 추가적인대사과정을거치는다른아미노산과달리 serine과 glycine은이후의대사과정을거치지않으며그자체로표피의보습유지에주요한역할을담당한다. 24,36,37 UV+ 군에서 serine과 glycine 및 alanine의함량은 UV-군과차이가없는반면, 피부보습은 UV-군에비해유의적으로낮았다. 이결과는자외선이투과성피지막을구성하는주요지질성분의함량을감소시킨다는사전연구와함께 38,39 자외선에의한피부보습함량이낮은이유는자연보습인자인유리아미노산보다는피부장벽을통해보습을유지하는지질메커니즘에의한것으로여겨진다. 추후자외선에따른피지막의주요구성성분인지질, 그중에서특히 ceramide 함량감소확인및녹차추출물의효능에대한연구가필요할것이라고사료된다. UV+1%GTE 군에서 serine과 glycine, alanine의함량은 UV+ 군및 UV- 군보다현저히높게나타났는데, 이는 serine과 glycine, alanine이피부보습에대한효과를직접적으로나타내는자연보습인자로써녹차섭취가자외선에의해야기되었던피부건조를완화시켰음을의미한다. Filaggrin으로부터분해된유리아미노산중 glutamate와 histidine은각각 PCA와 urocanic acid (UCA) 로빠르게대사되어자연보습인자의역할을수행하며, 40,41 arginine은 urea cycle의중간물질로작용한다. 36 따라서, glutamate, histidine 및 arginine으로는자외선조사및 GTE 의섭취효과를절대적으로반영하는데에는적절하지않을것으로사료된다. asparagine과 tryptophan은선행연구를통해 isoleucine, leucine, phenylalanine, threonine, tryptophan 등과같이 미량아미노산으로존재한다고보고되어 36 피부보습에있어서는큰영향을미치지않을것으로여겨진다. 위결과를종합해보면, 장기간의자외선조사는 profilaggrin 및 filaggrin, 유리아미노산의대사보다는 ceramide 를포함한피지막구성성분의변화와함께피부보습을감소시키는반면, 녹차추출물의식이공급은자외선대조군인 UV+ 군에비해표피 (pro)filaggrin, PAD3, 주요유리아미노산의발현변화를유의적으로증가시켜자외선조사로인한피부건조화를개선하였다. 요 본연구에서는자외선조사를병행하여녹차추출물의식이공급을 10주간공급한무모생쥐 (UV+1%GTE군) 의표피보습과 filaggrin 의대사물질및관련효소인 PAD3 의발현및함량변화를정상대조군 (UV-군 ) 및자외선조사군 (UV+ 군 ) 과비교분석하였으며결과는다음과같다. UV+ 군의피부보습은 UV-군보다유의적으로낮은반면 UV+1%GTE군의피부보습은 UV+ 군에비해유의적으로높았고, UV-군에는미치지못하였다. UV+ 군의 profilaggrin 함량은 UV-군과유의적인차이가없었으나, filaggrin의함량은 UV-군에비해유의적으로낮았다. UV+1%GTE군에서 profilaggrin은 UV- 및 UV+ 에비해현저히높았고, filaggrin은 UV-군과유사한수준으로높았다. UV+ 군의 PAD3 효소발현은 UV-군에비해유의적으로낮았다. UV+1%GTE군은 UV+ 군에비해유의적으로높았으며, UV-군에비해높았다. 자연보습인자의주요아미노산인 serine 및 glycine, alanine의함량은 UV-군과 UV+ 군간유사하였으나 UV+1%GTE군에서는 UV- 및 UV+ 군에비해현저히높았다. UV-군과비교하여 UV+ 군의피부보습함량이낮게측정된것에비해유리아미노산함량에는변화가없어자외선에따른피부보습에관여되는또다른메커니즘인표피지질함량분석과함께녹차추출물의효과에대한연구가추후이루어질필요성이있다. 그러나, 결론적으로자외선조사와병행된녹차추출물식이공급은 (pro)filaggrin의함량및 PAD3의발현이높게나타남에의한표피유리아미노산의함량이높아궁극적으로자외선조사에의해야기된피부건조를완화시켰다. 약 References 1. Rawlings AV, Harding CR. Moisturization and skin barrier function. Dermatol Ther 2004; 17 Suppl 1: 43-48.
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