RESEARCH ARTICLE Kor. J. Aesthet. Cosmetol., 단삼과황금의추출방법을달리한세포증식효과및항산화활성효과연구 전경미 *, 박경희, 표애자조선대학교대체의학과 A Research on Cell Proliferation Effect and Antioxidant Activity of Extracts Based on Different Extraction Methods of Salvia miltiorrhiza Bunge and Scutellaria baicalensis Kyung-Mi Jeon*, Kyeong-Hee Park, Ae-Ja Pyo Department of Complementary & alternative Medicine, Graduate School of Chosun University Abstract This research was conducted to identify industrial application potentials of functional cosmetics by suggesting the most effective method among the extraction method of active ingredients of natural products. The study used Salvia miltiorrhiza bunge and Scutellaria baicalensis as natural products and extracted the active ingredients using the hydrothermal, distillation, organic solvents (ethanol), ultrasonic waves (ethanol) and ultrasonic waves (water) extraction methods. The study filtered 10 types of extracts three times with the 20 μm filter and separated the supernatants (5,000 rpm, 5 min) using the centrifuge. The study identified the effects of cell proliferation of each extract using MTT assay and researched the antioxidant status through the DPPH radical scavenging activity test. The cell strains used in the MTT assay used HaCaT keratinocytes. The DPPH test was carried out in the method by Lu and Foo and measured the absorbance at 517 nm. Research result In terms of the cell proliferation effect through MTT assay, the research discovered that the ultrasonic (ethanol) extracts had the highest effect of 142% in Salvia miltiorrhiza bunge and 134.5% in Scutellaria baicalensis, indicating that the extraction method was the most suitable one for cell proliferation. When it comes to the DPPH radical scavenging activity, all extraction methods of two natural products had a high antioxidant status over 50%. By natural product type, the antioxidant status was very high in ethanol extracts (90.3%), ultrasonic waves (water) extracts (86.9%) of Salvia miltiorrhiza bunge and ethanol extracts (87.8%) and ultrasonic waves (water) extracts (87.4%) of Scutellaria baicalensis, demonstrating that ethanol and ultrasonic waves (water) extraction methods are the most suitable methods. Such research result was identified to have industrial application potentials such as developing functional cosmetics and be useful to develop anti-hair loss products. Keywords: Salvia miltiorrhiza bunge, Scutellaria baicalensis, Ultrasonic Waves Extraction Method, Antioxidant Status, Hair Loss 화장품원료로천연물을활용한역사는오래되었는데, 일본 에서는예부터미강 ( 米糠 ) 이나수세미물등을이용하여피부를 가꿔왔고, 중국에서는 외태비요 ( 外台秘要 ), 천금익방 ( 千金瀷 方 ) 등에나타나있는바와같이이미약초를화장품으로이용 *Corresponding author: Kyung-Mi Jeon, Department of of Complementary&alternative Medicine, Graduate School of Chosun University, 309 Pilmun-daero Dong-gu, Gwang-ju 501-759, Republic of Korea Tel.: +82 62 220 3052, Email: skinshop4296@naver.com Received June 16, 2015; Revised July 31, 2015; Accepted August 13, 2015; Published August 30, 2015 www.kosac.or.kr 495
Kor. J. Aesthet. Cosmetol., 하고있었다. 우리나라에서도화장품의주요원료를수입하던것에서탈피하여우리나라여성들의피부에적합한한방이나생약성분의신소재를개발하여제품화하는움직임이활발히일고있으며, 최근천연물에서유래된기능성제품의개발이나연구가지속적으로진행되고있다 (Im, 2012). 단삼 (Danshen, Salvia miltiorrhiza Bunge) 은꿀풀과 (Labiatae) 에속하는다년생본초인동속 ( 同屬 ) 근록식물 ( 近緣植物 ) 의근 ( 根 ) 으로고 ( 苦 ), 미한 ( 微寒 ), 무독 ( 無毒 ) 하고, 활혈조경 ( 活血調經 ), 랑혈소옹 ( 涼血消癰 ), 양혈안신 ( 養血安神 ) 등의효능이있어월경불순, 혈체경폐 ( 血滯經閉 ), 산후복통, 징하적취 ( 癥瘕積聚 ) 등의병증을치료하는것으로알려져있다. 중국이원산지로서드물게는약용으로재배하고굵은뿌리의표피에적색색소가침착되어있다. 단삼은피를맑게하여혈액순환에효능이있어혈액응고를방지하고혈액순환을촉진시켜혈전형성을억제시키는생약으로한방에서부인의생리불순, 생리통, 산후복통및어혈의심복부동통, 타박상의치료와불면증, 피부발진등의약재로쓰인다 (Kim, 1989). 단삼의주요화학성분으로는 tanshinone Ⅰ, ⅡA, ⅡB등을포함하는 diterpene 화합물과 danshensu, protocatechuic aldehyde, salvianolic acid B 등을포함하는 phenolic 화합물들, 그외에 baicaline, β-sitosterol, ursolic acid, vitamin E 와 tannin 등이알려져있으며 (Fugh-Berman, A, 2000), 단삼의밝혀진생리활성으로는항균 (Mok et al., 1994; Mok et al., 1995; Choi & Han, 2003), 항산화 (Kang et al., 2003), 항암 (Kim et al., 1999), 항돌연변이 (Ahn et al., 1999), 단삼유래 magnesium salvianolate의간염에효과 (Hase et al., 1997), 항혈전효과 (Yang et al., 2007) 등이있다. 황금 (Scutellaria radix) 은꿀풀과 (Labiatae) 에속하는다년생초본으로서황금 ( 속썩은풀, Scutellaria baicalensis Georgi) 의주피를벗긴뿌리를말한다. 뿌리에는 35% 이상의페놀화합물을포함하며노란색을띄기때문에황금 ( 黃芩 : golden root) 이라한다 (Song, 1981). 황금은전세계적으로약 300여종에이르는데, S, rivularis, S. discoloer, S. indica, S. barbata 및 S. scadens 등잘알려진종을이용하여유효성분들을추출하기위한연구가수행되었다 (Joshee et al., 2002). 주성분은 1930년황금뿌리에서 wogonin이분리된이후현재까지 baicalin, baicalein 및 wogonin을비롯하여 50 여종의 flavones가분리정제되었다 (Kim et al., 2013; Tang & Eisenbrand, 1992; Zhou et al., 1997). 황금은강력한약리작용을발휘하는작물로알려져있으며, 소염, 진통, 항알러지, 항균, 항바이러스, 항암및산화방지효과를위한대체의학의재료로활용되어왔다 (Broncel, 2007; Burnett et al., 2007; Chou et al., 2003; Taira et al., 2004). 오래전부터황금은민간처방약재로사용되었을뿐만아니라한방에서는뿌리를말린것을이질 (dysentery), 발열 (pyrexia), 황달 (jaundice) 및옹 (carbuncle) 의치료제로사용되고있다 (Huang et al., 2005; Lam et al., 2000). 우리나라, 중국, 일본등에서황금을항산화효과와함께소염성해열, 진통, 지사, 염증성결막염, 두통, 위염, 장염등에사용되어왔으며최근에는고혈압, 동맥경화, 피부질환, 불면, 감기, 상습유산에도사용되고있다 (Young, 2004). 또한한의학에서황금은청열조습 ( 淸熱燥濕 ) 의효능을가진약물로분류되어폐열해수 ( 肺熱咳嗽 ), 습열사리 ( 濕熱瀉利 ), 황달 ( 黃疸 ), 열림 ( 熱淋 ), 토뉵 ( 吐衄 ), 붕루 ( 崩漏 ), 태동불안 ( 胎動不安 ), 옹종 ( 癰腫 ), 정창 ( 疔瘡 ), 목적종통 ( 目赤腫痛 ), 장열번갈 ( 壯熱煩渴 ) 등을치료하여급만성호흡기질환, 폐렴, 비염, 인후염, 감모등에응용될수있는것으로알려져있다 ( 전국한의과대학공동교재편찬위원회, 1991; 吳普等 1996; 國家中醫藥管理局, 1999; Park, 2010). 연구에따르면황금에는활성성분으로 baicalein, biacalin, chrysin, oroxylin-a, wogonin 및 wogonoside 등과같은 30 여종이상의 flavonoid가함유되어있으며, 그중 baicalin이가장많이함유되어있다. 항염증, 항당뇨, 항균, 항알레르기, 항바이러스, 항고혈압및항산화등과같은많은약리학및생물학적활성을가지는것으로알려져있다 (Burnett et al., 2007; Choufani et al., 2011; Jung et al., 2012; Lu et al., 2011). 연구에의하면황금에함유된페놀화합물인 baicalin, baicalein, wogonin 그리고 oroxylin A가염증발현에관여하는 COX-2 (cyclooxygenase-2) 및 NO (nitric oxide) 형성에관여하는 inos (inducible nitric oxide synthase) 를억제하는것으로나타났으며 (Chena et al., 2001), 염증유발인자인 TNF를억제함이확인되었다 (Krakauer et al., 2001). 또한황금은 2,000-4,000 ng/g의멜라토닌을함유하고있고멜라토닌은항산화, 항노화효과뿐아니라골아세포의분화및골성장을촉진하는것으로보고되었다 (Jerome & Kim, 1999). 식물로부터유용성분을획득하기위한다양한연구가수행되고있으며, 열수추출법, Soxhlet법, 고온용매추출법및기계적압착법등의전통적인방법이주로사용되고있다 (Cushnie & Lamb, 2005; Knorr et al., 2004). 전통적인추출법은수율이낮고, 열에의해변성될수있으며, 추출용매의소모량이많아비용이높아진다는단점을가지고있다. 이에따라최근에는식물에서활성물질을추출하기위해초임계유체법 (supercritical fluid extraction), 초음파처리법 (ultrasoundassisted extraction; UAE), 효소추출법 (enzymatic extraction) 등의여러가지새로운추출기술들이개발되었다. 이중초음파추출법은비용이저렴하고간편하며효율적인추출기법으로천연물산업에서효율적으로활용되고있다 (Lee & 496 www.kosac.or.kr
단삼과황금의추출방법에따른세포증식및항산화효과연구 Row, 2006). 초음파는 20 khz 이상의높은주파수의음파로서, 초음파가용매를통과함에따라발생하는공동현상 (cavitation) 으로인하여매우높은에너지가발생하게된다. 높은에너지의충격으로인하여고압이발생하게되고, 발생된높은압력은세포벽및세포내부구조를쉽게파열시킨다. 또한초음파의기계적효과로인하여용매가세포로침투하게되어, 추출효율을증가시킨다 (Paniwnyk et al., 2001; Park et al., 2004). 초음파추출은전통적인추출법에비해열로인한유용성분의파괴및가용성분위주의추출등을방지할뿐만아니라추출시간을단축하고, 높은추출수율과생리활성효과를기대할수있는추출방법이다 (Kim & Kim, 2006; Park et al., 2004). 본연구에서는천연물중탈모방지및육모 양모효과가있는것으로알려진단삼과황금에서추출방법을열수추출법, 증류추출, 유기용매 ( 에탄올 ) 추출법, 초음파 ( 에탄올 ) 추출법, 초음파 ( 물 ) 추출법으로각기달리하여유효성분을추출한후, 각추출물에대하여 MTT assay와 DPPH 라디칼소거능활성실험을통하여기능성화장품등의산업적응용가능성을확인하기위하여수행하였다. 1. 실험재료및기기 1) 생약제재본연구에사용한단삼 (Danshen, Salvia miltiorrhiza bunge) 과황금 (Scutellaria radix) 은국내산으로경동시장에서건조상태로구입하여 powder형태로분쇄본실험에사용하였다. 2) 세포주본연구에서사용한세포주는피부각질형성세포주 (HaCaT keratinocytes) 를 ATCC (USA) 사에서구입사용하였다. 2. 실험방법 1) 생약추출방법본연구에서는단삼과황금 2종의생약제재각각에대하여열수추출법, 증류추출법, 에탄올추출법, 초음파에탄올추출법, 초음파물추출법등 5종의추출법을사용하여추출한 10종의추출물을 20 μm filter로 3차여과후원심분리기 (Sorvall, USA) 를이용하여상등액을분리 (5,000 rpm, 5 min) 사용하였다. (1) 열수추출분쇄한천연물 2종에대하여각각 100 g에 10배 (w/v) 의증류수를가하여 100 로 4h 동안추출하였다. 추출액을 20 μm filter로 3차여과후원심분리 (5,000 rpm, 5 min) 하여상등액을분리냉장보관사용하였다. (2) 증류추출분쇄한천연물 2종에대하여각각 100g을반응조하부에넣고 10배 (w/v) 의증류수를부어서 impellar, 반응조하부, 반응조상부, 냉각관 ( 직류, 환류 ), 분액여두를설치하고약 60min 동안약재를불린후 105 로 120min간전탕하여찌꺼기를따로분리한다. 전탕액을무균실에있는증류추출기에넣고 107 로추출한다음 PYREX병에받는다. 받아진약침액은무기염류를침강시키기위해서하루동안냉장보관한다. 무기염류를침강시킨약침액은무기염류를제외한상층액을분리하였다. 상층액을 20 μm filter로 3차여과후원심분리상등액을냉장보관사용하였다. (3) 유기용매 ( 에탄올 ) 추출분쇄한천연물 2종에대하여각각 100g을비이커에넣고재료가충분히잠기도록에탄올 70% 용액으로실온에 48h 침지시켜여과한후, 추출액을감압농축기 (SB-1000, EYELA, Tokyo, Japan) 로농축하였고, 농축액을 20 μm filter로 3차여과후원심분리하여상등액을분리냉장보관사용하였다. 3) 사용시약및기기실험에사용된시약은 1.1-diphenyl-2-picrylhydrazyl(DP PH, Sigma chemical Co., USA), Xanthine, Xanthine Oxidase, Butylated hydroxyanisole (BHA), 2,2 -Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), Pyrogallol, Mushroom tyrosinase, L-3,4- dihydroxyphenyl-alanine (L-DOPA) 을구입하여사용하였으며, 그외의기타시약은특급시약을사용하였다. 실험에사용된기기는 UV/vis spectrophotometer (Hitachi 200-10, Japan), ELISA reader, Centrifuge, 회전농축기, 냉동건조기등을사용하여측정하였다. (4) 초음파 ( 에탄올 ) 추출초음파에탄올추출은초음파분산기 (Sonics VC750, USA) 를사용하여수행하였다. 건조분쇄한천연물 2종각각에대하여 100 g에 10배 (w/v) 의 70% 에탄올을첨가하고, 40 에서 40 khz의초음파로 1h 추출후 1h 간격으로 3회추출하였다. 추출액을감암농축기를사용하여농축한후 20 μm filter로 3차여과하였으며여과액은원심분리한후상등액을분리냉장보관사용하였다. (5) 초음파 ( 물 ) 추출초음파저온추출은초음파분산기를사용하여, 건조분쇄된 www.kosac.or.kr 497
Kor. J. Aesthet. Cosmetol., 천연물 2종에대하여각각 100 g에 10배 (w/v) 의증류수를첨가하고, 40 khz의초음파로 1h 추출후 1h 간격으로 3회추출하였다. 추출액을 20 μm filter로 3차여과하였으며여과액은원심분리한후상등액을분리냉장보관사용하였다. 2) 세포생존율측정 - MTT assay 세포생존율측정은 Mosmann (1983) 의방법에따라, 피부각질세포주인 HaCaT CELL 96 well plate에 5Χ10⁴ cells/ well의밀도로분주한다음 37, 5% CO₂ incubator에서배양시킨후각시료를농도별로처리하여 24h 배양하였다. 배양이끝나기 4h 전에 MTT[3-(4,5-dimethylthiazol-2yl)-2,5- diphenyltetrazoliumbromide]solution을각 sell에 100 μl씩넣고 4h 배양한다음상층액을제거하고형성된 formazan 을 DMSO (dimethylsulfoxide) 용액 100 ul로녹여서 microplate reader(molecular Devices, USA) 를이용하여 540 nm에서흡광도를측정하였다. 측정은시료용액의첨가구와무첨가구간의흡광도차이를백분율로나타내었다. 3) DPPH radical 소거능측정 96 well plate에에탄올에용해시킨 DPPH (Sigma) 용액 180 μl와 DMSO 용액에실험농도로용해한커큐민과 95% 추출한천연추출물 2종을각각 20 µl씩혼합한후차광상태에서 20min간반응시켰다. 반응이완료되면 microplate reader (Biotek, USA) 를이용하여 517 nm에서흡광도를측정하였다. 표준곡선은 ascorbic acid (Sigma) 를이용하였고, DPPH radical 소거활성을산출하였다 Inhibition (%) = [(A0-A1)/A0] 100 (A0= 대조군흡광도 A1= 실험군흡광도 ) 3. 통계처리 MTT assay와 DPPH 실험은각각독립적으로 4회반복실시하였다. 통계분석은SPSS(Package for Social Science, SPSS Inc., Chicago, IL, USA) 를이용하여평균 (mean)± 표준편차 (standard deviation) 로표시하였고, 데이터의 normality 와 homogeneity는 ANOVA test (one-way analysis of variancetest) 를확인하였고, 실험구간의차이는 p<.05 수준에서 one-way ANOVA와 Duncan s multiplerange test로평가하였다. Figure 1. MTT assay based on extraction method of natural substances (%). 1. MTT assay 결과 2 종의천연물에대하여추출방법을달리한추출물의 MTT assay 결과를 (Table 1, Figure 1) 에나타내었다. 시험결과에따르면 MTT assay 을통한세포증식효과는단 삼추출물에서황금추출물보다우수하게나타났으며, 단삼과황 금추출물모두에서초음파 ( 에탄올 ) 추출물이각각 142%, 134% 로매우높게나타났다. 이것으로 2 종의천연물에대한가장좋 은세포증식효과를위한추출방법은초음파 ( 에탄올 ) 추출방식 임을알수있었다. 또한단삼의경우증류추출물에서 93.3% 로 세포증식효과가높게나타나다음으로적합한추출방식임을 알수있었다. 1) 단삼추출물의 MTT assay 추출방법에따른단삼추출물의세포증식효과는초음파 ( 에탄 올 ) 추출물 142.2%, 증류추출물 93.9% 에탄올추출물 56.8%, 초음 파 ( 물 ) 추출물 28.2%, 열수추출물 13.8% 의순으로나타나초음파 ( 에탄올 ) 추출물의세포증식효과가가장높게나타났다 (Figure 2). 2) 황금추출물 MTT assay 추출방법에따른황금추출물의세포증식효과는초음파 ( 에탄올 ) 추출물 134.5%, 초음파 ( 물 ) 추출물 49.1%, 에탄올추출물 46.8%, 증 류추출물 32.1%, 열수추출물 16.3% 의순으로나타나초음파 ( 에탄 올 ) 추출물의세포증식효과가가장높게나타났다 (Figure 3). Table 1. MTT assay based on extraction method of natural substances (%) ( ) ( ) 13.8 0.027** 93.3 0.433** 56.8 0.003*** 142.2 0.013*** 28.2 0.079** 16.3 0.029** 32.1 0.217** 46.8 0.006*** 134.5 0.028*** 49.1 0.185** Data are means SD of 4 experiments. *p<.05, **p<.01, and ***p<.001 498 www.kosac.or.kr
단삼과황금의추출방법에따른세포증식및항산화효과연구 Figure 2. MTT Assay Result Depending on the Extraction Method of salvia miltiorrhiza bunge. Figure 4. Measurement of the DPPH-Radical Scavenging Activity on extraction method of natural substances (%) Figure 3. MTT Assay Result Depending on the Extraction Method of Scutellaria Baicalensis. Figure 5. Measurement of the DPPH-Radical Scavenging Activity Having Different Salvia miltiorrhiza Bunge Extraction Techniques. 2. DPPH radical 소거활성측정단삼과황금 2종의천연물을추출방법을달리한추출물에대하여 DPPH radical 소거활성법을이용하여항산화활성을측정하였다. 실험결과단삼과황금모두에서에탄올추출물이각각 90.3%, 87.9% 로가장높게나타나항산화활성효과를위한가장좋은추출방법임을알수있었다. 또한 2종의천연물추출방법모두에서 50% 이상의높은항산화활성효과가나타났다 (Table 2, Figure 4). 의순으로에탄올추출물의항산화능이가장높게나타났다 (Figure 5). Yang et al. (2007) 은단삼메탄올추출물의 DPPH는사용된범위내에서농도의존적으로 radical-scavenging 효과를나타냈으며, RC 50 값은 18.71 μg /ml로비교적높은항산화효과를나타낸다고보고하였다. Kim et al. (1998) 은연구에서단삼열수추출의전자공여능은 77.11% 로나타났다고보고하였다. 이는본연구의결과와일치한다는것을알수있다. 1) 단삼추출물의 DPPH radical 소거활성측정추출방법에따른단삼추출물의항산화활성효과는에탄올추출물 90.3% 로가장높게나타났으며, 초음파물 52.6%, 증류추출 69.8%, 열수추출물 66.1%, 초음파 ( 에탄올 ) 추출물 62.9% 2) 황금추출물의 DPPH radical 소거활성측정추출방법에따른황금추출물의항산화활성효과는에탄올추출물 87.8%, 초음파 ( 물 ) 추출물 87.4% 로비슷하게나타났으며, 열수추출물 74.7%, 증류추출물 73.2%, 초음파 ( 애탄올 ) 추출물 Table 2. Measurement of the DPPH-Radical Scavenging Activity on extraction method of natural substances (%) ( ) ( ) 66.1 0.775* 69.8 0.554** 90.3 0.125** 62.6 1.785* 86.9 0.533** 74.7 0.985* 73.2 0.274** 87.8 0.435** 66.4 2.378* 87.4 0.270** Data are means SD of 4 experiments. *p<.05, **p<.01, and ***p<.001 www.kosac.or.kr 499
Kor. J. Aesthet. Cosmetol., Figure 6. Measurement of the DPPH-Radical Scavenging Activity Having Different Scutellaria Baicalensis. Extraction Techniques 의순으로높게나타났다 (Figure. 6). Park & Kim (2008) 은황금에탄올추출물의전자공여능은 1,000 μg/ml 에서 72.3% 였다고보고하였다. Kim (2014) 은황금 70% ethanol 추출물 0.3% 고농도처리 하였을경우 DPPH 라디컬소거능이 95% 로높은소거율을모 였다고하였다. 이는본연구결과와일치하는것으로황금의항 산화효과가높다는것을알수있었다. 본연구에서는탈모방지및육모 양모효과가있는것으로 알려진단삼과황금에대하여추출방법을열수추출법, 증류추 출, 유기용매 ( 에탄올 ) 추출법, 초음파 ( 에탄올 ) 추출법, 초음파 ( 물 ) 추출법으로각기달리하여유효성분을추출한후, 각추출 물에대하여 MTT assay 와 DPPH 라디칼소거능활성실험을 하였다. 주요연구결과는다음과같다. 세포증식효과는 2 종모두에서초음파 ( 에탄올 ) 추출물이단삼 142%, 황금 134.5% 로매우높게나타나초음파 ( 에탄올 ) 추출법 에의한유효성분추출이가장효과적임을알수있었다. DPPH radical 소거능효과는단삼과황금의모든추출방법 에서 50% 이상으로높게나타났다. 천연물종류별로는단삼의경우에서에탄올추출물 (90.3%), 초음파 ( 물 ) 추출물 (86.9%), 황금은에탄올추출물 (87.8%), 초음 파 ( 물 ) 추출물 (87.4%) 로매우높게나타나가장좋은방법임을 알수있었다. 이러한연구결과단삼과황금추출물의기능성화장품등의 산업적응용에서가장효율적인추출방법을선택하는자료로 활용가능하다는것을알수있었다. 전국한의과대학공동교재편찬위원회. 본초학. 영림사, 서울, 178-179, 1991. 吳普等述, 孫星衍, 孫馮翼輯. 神農本草經. 科學技術文獻出版社 ; 北京, p64, 1996. 國家中醫藥管理局 中華本草 編委會. 中華本草. 上海科學技術出版社 ; 上海, pp200-210, 1999. Ahn BY, Kim DG, Choi DS. Antimutagenic effect of tansen (Salvia miltiorrhiza Bunge). Kor. J. Appl. Microbil. Biotechnol., 27: 197-202, 1999. Broncel M. Antiatherosclerotic properties of flavones from the roots of Scutellaria baicalensis georgi. Wiad. Lek., 60: 294-297, 2007. Burnett BP, Jia Q, Zhao Y, Levy RM. A medicinal extract of Scutellaria baicalensis and Acacia catechu acts as a dual inhibitor of cyclooxygenase and 5-lipoxygenase to reduce inflammation. J. Med. Food, 10: 442-451, 2007. Chena YC, Shenb SC, Chena LG, Tony JF, Yang LL. Wogonin, baicalin, and baicalein inhibition of inducible nitric oxide synthase and cyclooxygenase-2 gene expressions induced by nitric oxide synthase inhibitors and lipopolysaccharide. Biochem. Pharmacol.. 6: 1417-1427, 2001. Choi HY, Han YS. Isolation and identification of antimicrobial compound from Dansam(Salvia miltiorrhiza Bunge). J. Food. Sci. Nutr., 32: 22-28, 2003. Chou TC, Chang LP, Li CY, Wong CS, Yang SP. The antiinflammatory and analogesic effects of baicalin in carrageenan-evoked thermal hyperalgesia. Anesthesia and Analgesia, 97: 1724-1729, 2003. Choufani E, Diligent J, Galois L, Mainard D. Metastatic renal cell carcinoma presenting as foot metastasis: case report and review of the literature. J. Am. Podiatr. Med. Assoc., 101: 265-268, 2011. Cushnie TP, Lamb AJ. Antimicrobial activity of flavonoids. Int. J. Antimicrob. Agents., 26, 2005. Fugh-Berman A. Herbs and dietary supplements in the prevention and treatment of cardiovescular disease. Prev. Cardiol., 3: 24-32, 2000. Hase K, Kasimn R, Basnet P, Kadota S, Namba T. 500 www.kosac.or.kr
단삼과황금의추출방법에따른세포증식및항산화효과연구 Preventive effect of lithospermate B from Salvia miltiorrhiza on experimental hepatitis induced by carbon tetrachloride or D-galactosamine/ lipopolysaccharide. Planta Med., 63: 22-26, 1997. Huang Y, Tsang SY, Yao X, Chen ZY. Biological properties of baicalein in cardiovascular system. Curr. Drug. Targets Cardiovasc. Haematol. Disord., 5: 177-184, 2006. Im DY. Volatile Compounds Analysis of Essential Oil Extracted from Dried Taraxacum coreanum. Kor. J. Aesthet. Cosmetol., 10: 797-801, 2012. Jerome AR, Kim BG. Melatonin Promotes Osteoblast Differentiation and Bone Formation. J. Biol. Chem., 274: 22041-22047, 1999. Joshee N, Patrick TS, Mentreddy RS, Yadav AK. Skullcap: Potential medicinal crop. Trends in new crops and new uses. Janick J and Whipkey (eds.) A. ASHS Press Alexandria., VA: 580-586, 2002. Jung HS, Kim MH, Gwak NG, Im YS, Lee KY, Sohn Y, Choi H, Yang WM. Antiallergic effects of Scutellaria baicalensis on inflammation in vivo and in vitro. J. Ethnopharmacol., 141: 345-349, 2012. Kang HS, Chung HY, Byun DS, Choi JS. Further isolation of antioxidative (+)-1-hydroxypinoresinol- 1-O-β-D-glucoside from the rhizome of S a l v i a miltiorrhiza that acts on peroxynitrite, total ROS and 1,1-dipheneyl-2-picrylhydrazyl radical. Arch. Pharm. Res., 26: 24-27, 2003. Krakauer T, Li B, Young H. The flavonoid baicalin inhibits superantigen-induced inflammatory cytokines and chemokines. FEBS Lett., 500: 52-55, 2001. Kim DH, Kim HJ, Chung BW. Extraction of antioxidative substance from Haematococcus pluvialis using ultrasonification. J. Eng. Res., 37: 79-86, 2006. Kim GY, Kang WW, Choi SW. A study on the quality characteristics of Sulgidduk added with persimmons leaves powder. J. East Asian Soc. Dietary Life, 9: 461-466, 1999. Kim JK. Illustrated Natural Gruges Rncyclopedia, Namsandong Publishers, Seoul, Kore, p160, 1989. Kim NY. Effect of Antioxidation and Inhibition of Melanogenesis from Scutellaria baicalensis Extract. Kor. J. Aesthet. Cosmetol., 12, 41-47, 2014. Kim NY, Moon JS, Choe TB. Effect of Scutellaria Baicalensis Extract Metalloproteinase-1 on the Expression of UV-B-irradidated Human Dermal Fibroblast. Kor. J. Aesthet. Cosmetol., 11: 649-654, 2013. Kim SM, Cho YS, Kim EJ, Bae MJ, Han JP, Lee SH, Sung SK. Effect of Hot Water Extracts of Salvia miltiorrhizr Bge., Prunus Persica Stokes, Angelica gigas Nakai and Pinus strobus on Lipid Oxidation. J. Kor. Soc. Food Sci. Nutr., 27: 399-405, 1998. Knorr D, Zenker M, Heinz V, Lee DU. Application and potential of ultrasonics in food processing. Trends Food Sci. Technol., 15: 261-266, 2004. Lam TL, Lam ML, Au TK, Ip DT, Ng TB, Fong WP, Wan DC. A comparison of human immunodeficiency virus type-1 protease inhibition activities by the aqueous and methanol extracts of Chinese medicinal herbs. Life Sci., 67: 2889-2896, 2000. Lee KJ, Row KH. Enhanced extraction of isoflavones from Korean soybean by ultrasonic wave. Kor. J. Chem. Eng., 23: 779-787, 2006. Lu Y, Joerger R, Wu C. Study of the chemical composition and antimicrobial activities of ethanolic extracts from roots of Scutellaria baicalensis Georgi. J. Agric. Food Chem., 59: 10934-10942, 2011. Mok JS, Kim YM, Kim SH, Chang DS. Antimicrobial property of the ethanol extract from Salvia miltiorrhiza. J Fd Hyg Safety., 10: 23-28, 1995. Mok JS, Park UY, Kim YM, Chang DS. Effect of solvents and extracting condition on the antimicrobial activity of Salvia miltiorrhiza radix(salvia miltiorrhiza) extract. J. Food Sci. Nutr., 23: 1001-1007, 1994. Paniwnyk L, Beaufoy E, Lorimer JP, Mason TJ. The extraction of rutin from flower buds of Sophora japonica. Ultrason Sonochem., 8: 299-303, 2001. Park CS, Kim DH. Biological Activities of Extracts from Scutellaria baicalensis, Zizyphus jujuba and Atractylodes macrocephala. Kor. J. Herbology., 23: 41-51, 2008. Park JH, Lee HS, Mun HC, Kim DH, Seong NS, Jung HG, Bang JK, Lee HY: Improvement of Anticancer Activation of Ultrasonificated Extracts from Acanthopanax senticosus Harms, Ephedra sinica Stapf, Rubus coreanus Miq. and Artemisia capillaris www.kosac.or.kr 501
Kor. J. Aesthet. Cosmetol., Thunb. Kor. J. Med. Crop. Sci., 12: 273-278, 2004. Park WS. The Effect of Bacillus-Fermented Scutellariae Radix Acupuncture Solution on Interleukin Production in Mouse Macrophage Stimulated by Lipopolysaccharide. 경락경혈학회지, 27:95-106, 2010. Song WZ. Studies on the resource of the chinese herb Scutellaria baicalensis georgi. Acta. Pharm. Sin., 16: 139-145, 1981. Taira Z, Yabe K, Hamaguchi Y, Hirayama K, Kishimoto M, Ishida S, Ueda Y. Effects of Sho-saiko-to extraxt and its components, Baicalin, baicalein, glycyrrhizin and glycyrrhetic acid, on pharmacokinetic behavior of salicylamide in carbon tetrachloride intoxicated rats. Food Chem. Toxicol., 42: 803-807, 2004. Yang SA, Im NK, Lee IN. Effects of Methanolic Extract from Salvia miltiorrhiza Bunge on in vitro Antithrombotic and Antioxidative Activities. Korean J. Food Sci. Technol., 39: 83-87, 2007. Zhou Y, Hirotani M, Yoshikawa T, Furuya T. Flavonoids and phenylethanoids from hairy root cultures of Scutellaria baicalensis. Phytochem., 44: 83-87, 1997. Herbal medicine. Young-lim Sa, Seoul, pp218-220, 2004. 502 www.kosac.or.kr