RESEARCH ARTICLE 추출용매와부위별복령 (Poria cocos) 의항산화, 항주름, 항염증평가 장영아, 이진태 * 대구한의대학교화장품약리학과 The Evaluation of Antioxidant, Anti-inflammatory, and Anti-aging of Extract Solvent and Poria cocos by Parts Young-Ah Jang, Jin-Tae Lee* Department of Cosmeceutical Science, Daegu Haany University Abstract This study is for checking the possibility of Poria cocos as cosmetic materials. For this we carried out biological active evaluation about antioxidant, anti-inflammatory and anti-aging by using extraction parts of Poria cocos and solvent extraction. We extracted Poria cocos by extraction parts (bark, inside skin) and solvents (water extracts, ethanol), and then in order to evaluate antioxidant activity we processed samples by concentration and carried out DPPH radical scavenging and ABTS radical scavenging. Also, in order to evaluate effect of anti-aging we carried out evaluation of Elastase inhibitory activity and Collagenase inhibitory activity. To evaluate effect of anti-inflammatory we evaluated toxicity of samples through MTT assay with a macrophage (raw 264.7 cells) and measured Nitric Oxide production inhibitory activity. As a result at the experiment of DPPH radical scavenging and ABTS radical scavenging of antioxidant evaluation, ethanol extracts showed more superiority in all three groups, A, B and C than water extracts of Poria cocos of evaluations of the effect against wrinkles, according to the result of measurement of elastase inhibitory activity, PCE (A) showed 96.7% effect and PCE(B) showed 98.3% effect respectively at µg/ ml concentration. Also, according to the result of measurement of collagenase inhibitory activity, PCE (A) showed.% effect and PCE (B) showed 72.2% effect respectively, so it means effect of ethanol extracts is superior. According to the result of nitric oxide production inhibitory activity, PCW (A) showed.4% effect and PCW (B) showed.3% effect respectively. This showed that effect of anti-inflammatory was greater in water extracts. In other words, ethanol extracts of Poria cocos proved effective against wrinkles by hindering antioxidant capacity and activities of elastase and collagenase by ROS. Water extracts showed superiority of effect of anti-inflammatory, so it suggests that Poris cocos extracts can be used as natural substance of cosmetics which are safe in antioxidant, anti-inflammatory, and anti-aging. Keywords: Poria cocos, Antioxidant, Anti-inflammatory, Anti-aging 여성들의사회진출이활발해짐과동시에육체적, 정신적으 로건강한삶을추구하게되었으며삶의행복과만족도를높이 *Corresponding author: Jin-Tae Lee, Department of Cosmeceutical Science, Daegu Haany University, Gyungsan, 712-749, Republic of Korea Tel.: +82 53 819 143, E-mail: yaviol@nate.com Received May 4, 15; Revised June, 15; Accepted June 22, 15; Published June 3, 15 는것중하나로아름다움에대한욕구가증가함에따라피부미용에대한관심이증가하고있다 ( 황완균등, 1). 이에깨끗한피부를가지기위해다양한천연소재를이용한화장품의개발이이루어져천연화장품, 한방화장품등이기능성화장품의전성시대를맞고있다 ( 박은예와김영철, 9). 식물유래유효성분에는플라보노이드, 폴리페놀, 베타카로틴, 리코펜등다양한성분들이존재하며이들성분은인체내로흡수되어항산화, 항염, 항암, 피부진정등의효능을나타낸다 ( 정상철등, 13). 천연물유래소재의경우화학공정이배제되고피부친화력이강하기때문에안전하고자연친화적인제품으로인식되어화장 377
품개발에새로운트렌드가되고있다 ( 김도훈등, 1). 피부미용분야에는항산화, 주름개선, 탄력증가, 피부미백등에영향을미치는생물학적부분에특이적으로작용하여적은양으로높은기능을할수있는화장품, 피부미용소재에관심이집중되고있다 ( 김경동과김상진, 7; 이용규와조재열, 7). 사람의피부는노화과정에서다양한물리 화학적인변화가일어나며크게 2가지의원인중유전적요인과호르몬대사작용에의해발생하는내인성노화와자외선, 흡연, 오염등의환경적요인에의해발생하는외인성노화로나뉜다 ( 김아름등, 11). 자외선은파장에따라 UVC ( 2 nm), UVB (2 3 nm), UVA (3 nm) 의 3가지로나누어지며 UVC 는자외선영역중에서생물학적인손상을가장많이유발할수있지만, 대부분은지구대기의성층권에있는오존층에서흡수되어지표상에는거의도달하지않으며 UVA와 UVB는피부에작용하여활성산소종 (Oxidative Reactive Species, ROS) 을다량으로생성하여진피결합조직의손실과함께피부항상성을깨뜨린다 (Svobodova et al., 6; Brenneisen et al., 2). 피부세포는자외선흡수에의한손상을방어하기위해멜라닌 (melanin) 생성을촉진시켜기미, 주근깨, 잡티, 검버섯등의과색소침착 (hyperpigmentation) 을유발하여미용적인측면에서좋지않은인상을주기도한다. 뿐만아니라외부환경에의해증가된산화적스트레스는각질형성세포 (Keratinicyte) 의구조적인변화와함께염증유발사이토카인 (pro-inflammatory cytokine) 의분비를유도하여피부건조증, 가려움증을유발하고, 심한경우에는피부염으로악화될수있다 ( 심정기등, 9; Thong et al., 8; Wilmer et al., 1994). 현재피부효능이우수한화장품, 피부미용소재로널리알려진미백물질은알부틴 (arbutin), 비타민 C(ascorbic acid), 코직산 (kojic acid) 등이있으며주름개선으로쓰이고있는물질로는레티놀 (retinol), 레티닐팔미테이트 (retinyl palmitate), 아데노신 (adenosine) 등이사용되고있다 (Takizawa et al., 3). 이러한성분들은효과가뛰어난것으로알려져있음에도불구하고제품의안전성에서끊임없이문제가제기되고있다 ( 김보영등, 1). 따라서천연물로부터유래하는효능이우수하며피부자극완화효능이함께있는화합물에관한연구가활발하게수행되고있다. 복령 (Poria cocos) 은버섯류에속하며단단한균핵의내부색깔에따라육질이견고한백복령과연하고부드러운적복령으로구분되는데맛은달콤하고향과독이없다 ( 강안석등, 1999). 복령의주성분으로는탄수화물, 수분, 조섬유질, 무기질및미량의단백질등이함유되어있으며, 효능으로는위궤양예방, 이뇨작용, 심신의보양, 면역증강, 식욕증진, 뇌세포의활성등이보고되고있다 (Lee et al., 1982). 복령균사체에서분리한U-pachymaran, pachymaran, carboxymethyl pachymaran 등은항암효과가있으며, (1,3)-(1,6)-β -D-glucan은강한항종양성이있다고하였다 (Chihara G et al., 1971; 권미선등, 1998). 또한복령 (Poria cocos) 중의 triterpene 성분은항구토, 항염증, 항피부암에효과가있다고보고된바있다 (Nukaya et al.,1996). 상기와같이복령의유효성분규명및약리작용에관한연구에관해일부이루어졌으나화장품소재분야에서의항산화, 항염, 항주름에관한전반적인생리활성의특성연구는이루어지지않아복령의추출부위 ( 껍질, 내피 ) 및용매별 ( 열수, 에탄올 ) 추출을이용해화장품소재로서의가능성을알아보기위해연구하였다. 1. 추출물의제조본실험에사용된복령은경상북도경산시한국한방사업협동조합으로부터제공받아사용하였다. 복령의추출방법은껍질채로편썰기 (A), 껍질을분말로파쇄하기 (B), 내피를분말로파쇄하기 (C) 의방법으로용매를시료중량의 1배양을가하여실온에서 24 h 침지하여상징액과침전물을분리하여동일한방법으로 3회반복추출하였다. 천연복합물을원심분리및여과, 농축 (rotary vaccum evaporator, HS-1SP, Hanshin, Korea) 하여동결 (FD5525, Ilshin, Korea) 건조하였고냉장실에보관하여사용하였다. 2. Electron donating abilities 추출물의 DPPH radical activity (Electron donating abilities, EDA) 는다음과같이측정하였다. 각시료용액 2. ml에.2 mm의 1,1-diphenyl-2-picrylhydrazyl (DPPH).5 ml를넣고교반한후 3 min간방치한다음 517 nm 에서흡광도를측정하였다. 전자공여능효과는시료용액의첨가구와무첨가구의흡광도감소율로나타내었다. 3. ABTS + radical cation 소거작용측정 ABTS + radical cation 소거작용측정은 ABTS + radical cation은 7 mm ABTS와 2.4 mm potassium persulfate (K2S2O8) 을 1:1로혼합하여암실에서실온으로 24 h 동안반응하였다. 사용전에 ABTS + 용액을에탄올에희석하여 734 nm 에서흡광도값이.76±.1이되게하여사용하였다. 4. Elastase 저해활성측정 Elastase 저해활성은다음과같이측정하였다..4 M Tris- HCl buffer (ph 8.6) 을 5 µl, 농도별추출물을 5 µl, 5 mm Tris-HCl buffer (ph 8.6) 에.6 U/ml이되도록희석한 378
복령의화장품소재평가 Electron donating ability (%) enzyme (ex-porcine pancreas) 을 5 µl 을넣고 37 에서 3 min 간반응시킨후 substrate (methoxysuccinyl-l-ala-l- Val-5-nitroanilide) 를 µl 를넣은후 37 에서 3 min 간 반응시킨후다시 41 nm 에서흡광도를측정한다. 5. Collagenase 저해활성측정 Collagenase 저해활성은반응구.1 M Tris- HCl buffer (ph 7.5) 에 4 mm CaCl 2 를첨가하여, 4-phenylazobenzyloxycarbonyl- Pro-Leu-Gly-Pro-D- Arg (.3 mg/ml) 를녹인기질액.25 ml 및시료용액.1 ml 의혼합액에 collagenase (.2 mg/ml).15 ml 를첨가하여실 온에서 min 간방치한후 6% citric acid.5 ml 을넣어반응 을정지시킨후, ethyl acetate 1.5 ml 을첨가하여 3 nm 에 서흡광도를측정하였다. Collagenase 저해활성은시료용액의 첨가구와무첨가구의흡광도감소율로나타내었다. 6. 세포배양 마우스의대식세포주인 Raw 264.7 cell 은한국세포주은행 (KCLB) 에서분양받았으며세포배양을위해서항생제 ( U/ ml Penicillin, U/mL Streptomycin) 를첨가하고 FBS 가 1% 비율로함유된 DMEM 배지에서온도 37, 5% CO 2 조건 에서배양하였다. 7. 세포독성측정 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) BHA Figure 1. DPPH radical scavenging activity (%) of dependent on concentration from extracts of Poria cocos. PCW (A): cut thin slice extracted with water, PCW (B): grinded into powder extracted with water, PCE (A): cut thin slice extracted with 7% ethanol, PCE (B): grinded into powder extracted with 7% ethanol, PCE (C): grinded into soft powder extracted with 7% ethanol. Results are expressed as means±s.d. of triplicate data. 세포생존율측정은 MTT assay 로측정하였다. 세포를 96 well plate 에.6~8 1 5 cells/well 이되게.18 ml 분주하고, 시료를농도별로조제하여.2 ml 첨가한후 37, 5% CO 2 ABTS+ radical scavenging activity (%) incubator 에서 24 h 배양하였다. 대조군은시료와동량의증류 수를첨가하여동일한조건으로배양하였다. 여기에 5 mg/ml 농도로제조한 MTT 용액.2 ml 를첨가하여 4 h 배양한후 배양액을제거하고각 well 당 DMSO:Ethanol (1:1).15 ml 를가하여실온에서 3 min 간반응시킨뒤 ELISA reader 로 55 nm 에서흡광도를측정하였다. 세포생존율측정은시료용 액의첨가군와무첨가군의흡광도감소율로나타내었다. 8. Nitric oxide (NO) 저해능측정 Raw 264.7 cell 을 6 well plate 에배지 1 ml 당 1 1 5 개의 세포로배양하여 confluence 가 % 일때, PBS 로 2 번세척 한후무혈청배지를사용하여 12 h 이상배양한다음 LPS 1μ g/ml 을 control 군을뺀모든 well 에넣어서자극시켰다. 2 h 후에복령추출물을농도별로처리하고 NO 생성량은 24 h 후 에상층액을모아 griess regent 로 1 min 간반응시킨후에 ELISA reader 를이용하여 5 nm 에서흡광도를측정하였다. 1. Electron donating abilities 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) BHA Figure 2. ABTS+ radical scavenging activity (%) of dependent on concentration from extracts of Poria cocos. PCW (A): cut thin slice extracted with water, PCW(B): grinded into powder extracted with water, PCE (A): cut thin slice extracted with 7% ethanol, PCE (B): grinded into powder extracted with 7% ethanol, PCE (C): grinded into soft powder extracted with 7% ethanol. BHA : Butylated hydroxyanisole. Results are expressed as means±s.d. of triplicate data. DPPH 는비교적안정한 free radical 로써, ascorbic acid, tocopherol, polyhydroxy 방향족화합물, 방향족아민류에의 해환원되어짙은자색이탈색되는원리를이용하여항산화활 성을간단히측정할수있는동시에식물체의항산화활성과도 연관성이매우높기때문에많이이용되고있는방법이다 ( 장미 379
Elastase inhibition rate (%) Elastase inhibition rate (%) 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) VIT.C 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) VIT.C Figure 3. Elastase inhibition rate (%) of dependent on concentration from extracts of Poria cocos. PCW (A): cut thin slice extracted with water, PCW (B): grinded into powder extracted with water, PCE (A): cut thin slice extracted with 7% ethanol, PCE (B): grinded into powder extracted with 7% ethanol, PCE (C): grinded into soft powder extracted with 7% ethanol. VIT.C : ascorbic acid, Results are expressed as means±s.d. of triplicate data. Figure 4. Collagenase inhibition rate (%) of dependent on concentration from extracts of Poria cocos. PCW (A): cut thin slice extracted with water, PCW (B): grinded into powder extracted with water, PCE (A): cut thin slice extracted with 7% ethanol, PCE (B): grinded into powder extracted with 7% ethanol, PCE (C): grinded into soft powder extracted with 7% ethanol. VIT.C : ascorbic acid, Results are expressed as means±s.d. of triplicate data. 진등, 5). 복령추출물의 DPPH radical 소거능을측정한결과 Figure 1과같이나타내었다. 실험결과농도가높아질수록유의적으로활성이증가하였으며열수추출물대비에탄올추출물이 A, B, C 군모두에서효능이우수하였다. µg/ml 농도에서 PCE (C) 가 93.5% 로동일농도에서대조군 BHA 95.3% 와유사한효능을나타내었다. 2. ABTS + radical cation activity ABTS + radical 소거능을측정한결과는 Figure 2와같다. 복령열수추출물은 μg/ml의농도에서각각 A (99.4)%, B (96.3%), C (82.)% 의활성을나타내어 PCW (A) 시료가가장높은활성을나타내었다. 에탄올추출물은 μg/ml의농도에서각각 A (97.9)%, B (86.6%), C (73.9)% 의활성을나타내어 ABTS + radical 소거능결과는열수추출물의효능이더우수했으며 6개시료모두높은활성을나타내었다. ( 손미예, 7) 의연구에서발효한복령추출물이동일농도에서.% 의효능을나타낸것과비교했을때본실험에사용된열수, 에탄올추출물이활성이더높음을확인할수있었다. ABTS + radical 소거능의경우 hydrogendonating antioxidants와 chain breaking antioxidants 모두를측정할수있고, 친수성물질과소수성물질의항산화력측정에가능하므로일반적으로 DPPH 라디칼소거능보다는높은활성을나타낸다 (Re et al., 1999). 3. Elastase 저해활성측정피부탄력단백질인 elastin이 elastase에의해분해되어피부의그물망구조결합이끊어짐으로, elastase가주름생성의주 원인효소로알려져있다 (Tsuji et al., 1). Elastase 저해활성측정결과는 Figure 3에서보듯 µg/ml 농도에서 PCE (A), PCE (B), PCE (C) 가각 96.7%, 69.6%, 98.3% 로대조군 Vit.C의 % 와대비해서 PCE (A), PCE (C) 는유사한값으로높은저해율을나타내었고열수추출물은저해율이낮았다. 김정옥등 (8) 의상황버섯의열수, 에탄올추출물의 Elastase 저해율이동일농도에서각 22.2%, 31.5% 와비교했을때복령추출물이더활성이높음을확인하였다. 4. Collagenase 저해활성피부는크게표피, 진피, 피하지방층으로구성되어있다. 자연노화와광노화모두주름이생성되고피부면역세포인랑게르한스세포와진피의교원섬유가감소한다는공통점이있으나, 자연노화에서는피부가얇아지고, 광노화의경우에는탄력섬유의변성이심하며 collagenase 발현이증가되어있으므로, 피부가두꺼워지고변형된탄력섬유가증가하는특징을갖고있다 ( 이민호, 13; Waller & Maibach, 6). 따라서지금까지의주름개선연구는콜라젠을분해하는 collagenase, 젤라틴을분해하는 gelatinase의작용, 탄력섬유를분해하는 elastase 의작용을억제하는물질개발이중심이되어왔다 (Jung, 3; Mauviel et al., 1994). collagenase 저해활성실험결과추출물의농도 µg/ml의농도에서 PCE (A), PCE (B), 각.%, 72.2% 의효능을나타내었으며열수추출물은효능이없었으며시료의활성은 elastase 저해활성측정결과와유사하게나타났다. 또한윤우식등 (1) 의상황버섯에탄올추출물의 collagenase 활성억제가나타나지않은것과비교시복령이 3
복령의화장품소재평가 Cell viability (%) Nitrite production (μm) Con 1 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) ConN nnon 1 5 PCW(A) PCW(B) PCW(C) PCE(A) PCE(B) PCE(C) Figure 5. Cell viability of dependent on concentration of Poria cocos. PCW (A): cut thin slice extracted with water, PCW (B): grinded into powder extracted with water, PCW (C): grinded into soft powder extracted with water, PCE (A): cut thin slice extracted with 7% ethanol, PCE (B): grinded into powder extracted with 7% ethanol, PCE (C): grinded into soft powder extracted with 7% ethanol. The data represent the mean±sd of three separate experiments. 동일용매에서저해율이높음을알수있었다. 5. 세포생존률측정 복령열수, 에탄올추출물의항염증효능을알아보기위한 Raw 264.7 cell 의세포독성을확인한결과를 Figure 3 과같이 나타내었다. 복령열수추출물의세포생존율은 μg/ml 의 농도에서각각 A (93.6)%, B (94.5%), C (78.4)% 로세포독성 에영향을적게주는것으로나타났으며에탄올추출물은동일 농도에서각각 A (84.5)%, B (81.5%), C (69.)% 로 C 군이다 른군에비해 31% 의세포독성이나타났으며전체농도에서열 수추출물이에탄올추출물에비해세포독성에영향이없는것으 로나타났다. 6. Nitric oxide(no) 저해능 NO 는아질산염을측정하는 griess regent 를이용해간접적 으로측정할수있으며, 일산화질소 (nitric oxide, NO) 는높은 반응성을가진생체생성분자로서, 신경전달, 혈관의이완및 세포매개성면역반응에관여하는데, 생체의피부가염증상태 에빠지게되면 inducible NOS (inos) 가발현되어 NO 를생산 한다 (Choi et al., 2). 염증반응부위에서 NO 의과다발현은 과다염증및자가면역질환의매개물질로작용한다 (Southan & Szabo, 1996). 복령추출물의항염증효과를확인하기위해 NO 생성저해활성을측정한결과는 Figure 5 와같다. 생성된 NO 량을 Control 로하여 LPS 를 1 μg/ml 농도로처리한군을 % 로두고 LPS 를처리하지않은대조구와비교했을때 LPS 처리군에서 NO 량이상당히증가했음을알수있었고, 열수, 에 탄올의복령추출물을농도별로처리했을때모든시료에서농 Figure 6. Inhibitory effects of Poria cocos on the production of nitric oxide Raw 264.7 cells. Raw 264.7 cells were cultured with LPS (1 μg/ml) in the presence or absence of Poria cocos for 24 h to determine the level of NO. Con: LPS not induced group, Non: LPS induced group. The data represent the mean±sd of three separate experiments. 도가증가할수록 NO 생성량이뚜렷하게감소하는것을확인할 수있었다. 세포독성에기인하지않는데이터를위해서생존 률 9% 이상의시료인 PCW (A), PCW (B) 를처리한군의결과 를확인하였을때 LPS 단독군에비해 µg/ml 농도에서각.4%,.3% 의감소율을 5 g/ml 농도에서 21.4%, 25.8% NO 생성이감소함을볼수있었다. 본연구에서추출용매와부위별복령의항산화, 항주름, 항염 증실험을통해화장품기능성소재로서의가능성에대해평가 하여다음과같은결론을얻었다. 첫째, 항산화평가의 DPPH radical, ABTS + radical 소거 능실험모두복령의열수추출물대비에탄올추출물이 A, B, C 군모두에서효능이우수하였으며 DPPH radical 소거능결 과 µg/ml 농도에서 PCE (C) 가 93.5% 로가장높은효능을 나타내었고 ABTS radical 소거능결과 6 개시료모두대조군 BHA 와유사하게높은활성을나타내었다. 둘째, 주름효능평가중 Elastase 저해활성측정결과 µg/ml 농도에서 PCE (A), PCE (C) 가각 96.7%, 98.3% 로높 은저해율을나타내었고 Collagenase 저해활성측정결과동일 농도에서 PCE (A), PCE (B) 각.%, 72.2% 의효능을나타 내었으며주름효능평가실험에서는열수추출물의효능은매우 낮았으나에탄올추출물의효능은우수하였다. 셋째, 항염증효과를확인하기위해 NO 생성저해활성을측 정한결과 µg/ml 농도에서 LPS 단독군의 NO 생성량 % 에대비하여 PCW (A), PCW (B) 각.4%,.3% 의높 381
은감소율을보여열수추출물이항염증효능이우수함을확인하였다. 이상의연구결과로보아복령의에탄올추출물은 ROS에의한항산화능이있었으며복령의껍질, 내피에탄올추출물은 elastase, collagenase 활성을저해하여주름개선에효능이있는물질로나타났다. 또한열수추출물중껍질부분에서 Raw 264.7 세포의독성이없는범위내항염증효능이우수함을나타내었다. 따라서복령추출물은향후항산화, 항주름, 항염증에안전한화장품천연소재로의활용이가능함을시사한다. 강안석, 강태수, 손형락, 서상명, 강미선, 김광포, 이정숙. 복령의인공재배법개선과항산화활성에관한연구. 한국균학회지, 27: 378-382, 1999. 김경동, 김상진. 약용식물의항균효과및화장품응용에관한연구. 한국미용학회지, 13: 7. 김도훈, 이수진, 이현석, 오성근, 강학회, 김진웅. 최근화장품산업의신소재개발연구. 한국공업화학회지, 13, 11-23, 1. 김보영, 김타곤, 강환열, 백현, 전혜영, 김동욱. Porcine Placenta의기능성화장품소재특성. 화학공학회지, 48: 327-331, 1. 김아름, 김정은, 박수남. 상황버섯추출물의항산화및성분분석. 대한화장품학회지, 37: 39-318, 11. 김정옥, 정미정, 최향자, 이진태, 임애경, 홍주헌, 김대익. 상황버섯열수및에탄올추출물의항산화효과및생리활성. 한국식품영양과학회지, 37: 684-69. 8. 권미선, 정신교, 최종욱, 송경식, 강우원. 재배복령 (Poria cocos Wolf) 의채취시기에따른화학적품질과기능적특성. 한국식품영양과학회지, 27: 134-1, 1998. 박은예, 김영철. 백차열수추출물의피부미백유효성. 한국미용학회지, 15: 611-617, 9. 손미예. 버섯균사체로발효시킨복령과후박의항산화및항암효과식품산업과영양. 한국식품영양과학회지, 12: 51-57, 7. 심정기, 이지해, 양미숙, 서대방, 이상준. 콜라겐펩타이드의피부광노화예방효과. 한국식품과학회지, 41: 441-445, 9. 이민호, 김형진, 정현아, 이영근. 진피섬유모세포에서대복피추출물의세포외기질합성촉진효과. 한국산학기술학회논문지, 14: 1857-1862, 13. 이용규, 조재열. 대식세포주 RAW264.7 세포에서 curcumin의 lipopolysaccharide에의한 nitric oxide 생성억제효과. 한국약용작물학회지, 15: 451-456, 7. 윤우식, 정현아, 노석선. 상황버섯추출물이항염증및항알레르기, 항산화, 주름제거에미치는실험적연구. 한방안이비인후피부과학회지, 23: 75-93. 3. 장미진, 우미희, 김영호, 전도연, 이순재. Effects of antioxidative, DPPH radical scavenging activity and antithrombogenic by the extract of Sancho (Zanthoxylumschinifolium). 한국영양학회지, 38: 386-394. 5. 정상철, 박종호, 김진희. 천연소재로부터피부효능을갖는피부미용식품의개발동향. 대한피부미용학회지, 11: 675-684, 13. 황완균, 이영애, 안희진, 조연경. 여성의라이프스타일에따른화장품구매태도. 대한피부미용학회지, 8: 117-126, 1. Brenneisen P, Sies H, Scharffetter-Kochanek K. Ultraviolet-B irradiation and matrix metalloproteinases: from induction via signaling to initial events. Ann. NY. Acad. Sci., 973: 31-43. 2. Chihara G, Hamuro J, Yamashita Y, Ohsaka Y, Maeda Y. Carboxymethylpachmaran a new water soluble polysaccharide with marked antitumor activity. Nature, 233: 486-49, 1971. Choi BM, Pae HO, Jang SI, Kim YM, Chung HT. Nitric oxide as a pro-apoptotic as well as anti-apoptotic modulator. J. Biochem. Mol. Biol., 35: 116-126, 2. Jung JH. Photoaging in Asians. Photodermatol. Photoimmunol. Photomed., 19: 19-121. 3. Kanayama H, Adachi N, Togami M. A new antitumor polysaccharide from the mycelia of Poria cocos Wolf. Chem. Pharm. Bull., 31: 1115-1119, 1983. Lee KS, Lee MW, Lee JY. Studies on the antibacterial activity of Poria cocos. Kor. J. Mycol., 1: 27-31. 1982. Mauviel A, Halcin C, Vasiloudes, P, Parks WC, Kurkinen M, Uitto J. Unncoordinate regulation of collagenase, stromelysin, and tissue inhibitior of metalloproteinase genes by prostaglandin E2: selective enhancement of collagenase gene expression in human dermal fibroblasts in culture. J. Cell. Biochem., 54: 465-472, 1994. 382
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