• Title/Summary/Keyword: Biological screening methods

검색결과 123건 처리시간 0.029초

흑미(현진주벼)유래 안토시아닌의 항산화능 탐색 (Isolation of Anthocyanin from Black Rice (Heugjinjubyeo) and Screening of its Antioxidant Activities)

  • 박영삼;김선중;장효일
    • 한국미생물·생명공학회지
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    • 제36권1호
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    • pp.55-60
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    • 2008
  • 유색미는 회색 빛 외에 붉은색 흑은 보라색 계통의 색소를 겨 부분에 함유하고 있다. 특히, 흑미는 예로부터 건강식품으로 여겨져 왔다. 흑미(흑진주벼)는 최근에 anthocyanin포함하는 생리활성 물질원으로서 관심이 증대되고 있다. Anthocyanin은 수용성의 색소로서 flavonoid계통의 대표 색소로 알려져 왔다. 흑미에 함유되어있는 Anthocyanin으로는 cyanidin 3-O-glucoside와 peonidin 3-O-glucoside, malvidin 3-O-glucoside, pelagonidin 3-O-glucoside, delphinidin 3-O-glucoside등이 알려져 있다. 본 연구에서는, 흑미로부터 amberlite XAD7 chromatography방법을 사용해서 anthocyanin을 추출 후 HPLC와 UV-Vis spectrophotometer를 이용하여 정량분석 및 정성분석을 실시하였다. 그 결과 흑미의 anthocyanin 추출물 중 95%가 cyanidin 3-O-glucoside, 5%가 peonidin 3-O-glucoside임을 확인하였다. 그리고 in vitro상의 여러 가지 실험방법을 통하여 anthocyani의 항산화 효과를 측정하였다. 그 결과 100g/ml 농도의 anthocyanin추출물은 90%이상 linoleic acid emulsion의 과산화를 억제하였고, 55.20%의 DPPH 자유 라디칼 제거효과, 54.96%의 superoxide 자유 라디칼 제거효과, 72.67%의 과산화수소 제거효과를 보였다. 그리고 또한 높은 환원 능력을 확인할 수 있었다. 표준 항 산화 물질로는 BHA, BHT, ${\alpha}-tocopherol$을 사용하였다. 이 결과로 보아 anthocyanin추출물은 새로운 항산화 물질로서 이용될 수 있을 것이다.

Identification of Homer1 as a Potential Prognostic Marker for Intrahepatic Cholangiocarcinoma

  • Wu, San-Yun;Yu, Ming-Xia;Li, Xiao-Gai;Xu, Shu-Fang;Shen, Ji;Sun, Zhen;Zhou, Xin;Chen, Xing-Zhen;Tu, Jian-Cheng
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권7호
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    • pp.3299-3304
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    • 2014
  • Background: The aim of the present study was to analyze whether Homer1 is a potential prognostic marker for intrahepatic cholangiocarcinoma (ICC). Materials and Methods: The expression of Homer1 in ICC tissue was detected with immunohistochemistry and levels of protein in ICC and paratumor tissues were evaluated by Western blotting. Survival analysis by the Kaplan-Meier method was performed to assess prognostic significance. Results: Homer1 expression was high in 67.4% (58/86) of ICC samples, and there was significant difference between ICC and adjacent noncancerous tissues (p<0.001); high expression was associated with poor histologic differentiation (p=0.019), TNM stage (p=0.014), lymph node metastasis (p=0.040), and lymphatic invasion (p=0.025). On Kaplan-Meier analysis, a comparison of survival curves of low versus high expressors of Homer1 revealed a highly significant difference in OS (p=0.001) and DFS (p=0.006), indicating that high expression of Homer1 was linked with a worse prognosis. Multivariate analyses showed that Homer1 expression was an independent risk factor predicting overall survival[Hazard ratio(HR), 7.52; 95% confidence interval (CI), 2.63-21.47; p=0.002] and disease-free survival (HR, 11.56; 95%CI, 5.17-25.96; p<0.001) in ICC. Conclusions: Homer1 promotes lymphatic invasion and associates with lymph node metastasis and poor prognosis of ICC. The current study shows that Homer1 may be an independent prognostic factor for ICC patients after curative resection, and it provides an important basis for screening/treating high-risk patients.

202종(種) 한약재(韓藥材)의 $IFN-{\gamma}$ 분비조절에 관한 비교연구 (Comparative study of 202 herbs on interferon-gamma secretion)

  • 이시형;강미숙;최유경;전찬용;박종형;김동우
    • 대한한방내과학회지
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    • 제27권2호
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    • pp.336-344
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    • 2006
  • Objects : This study has been carried out to assess the effects of the variable herbs on $IFN-{\gamma}$ secretion in the mouse spleen cell. Methods : 202 kinds of herb extracts were used to evaluate the $IFN-{\gamma}$ secretory distinction by each $1{\mu}g/ml$ and $10{\mu}g/ml$ density of water. All experimental herbs were grouped by oriental herbalogical method. But each herb had its independent variables. Results : The secretions increased in 20% of all herbal water. The density differences also make different effects on the secretion of $IFN-{\gamma}$. The secretion of IFN-${\gamma}$ inclosed in some kinds of herbs of IFN-${\gamma}$. It has representatively increased in Imperaetae Rhizoma(白茅根) of $1{\mu}g/ml$ and Notopterygii Rhizoma(羌活)of $10{\mu}g/ml$. $IFN-{\gamma}$ incresed in 12 kinds of heybs of both densities. The secretion of $IFN-{\gamma}$ decreased in some kinds of herbs of $IFN-{\gamma}$. It has representatively decreased in Moutan Radicis Cortex(蘇丹皮) of $1{\mu}g/ml$ and Angelicae Radix(富歸尾) of $10{\mu}g/ml$. $IFN-{\gamma}$ decreased in 18 kinds of herbs of both densities. In t oriental herb group, The secretion of $IFN-{\gamma}$ increased in Bang-Hyang-Hwa-Sup group(芳香化濕藥), He-Pyo group(解表藥), I-Su-Sam-SuP group(利水渗) The secretion of $IFN-{\gamma}$ decreased in Gu-Chung group(驅蟲藥), An-Sin group(安神藥). Su-Sap group(收澁) On-Li group(溫裏藥), I-Gi group(利氣藥). Conclusions : The result of this study will not only broaden applications of oriental medicine to biological therapy, but also form the basis of oriental medical therapy to find out the meaning of oriental classification.

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Adjuvant 유발(誘發) 류마티스 관절염(關節炎)에 대한 소활락단(小活絡丹)의 억제(抑制) 효과(效果) (Suppressing Effects of Sowhalrack-dan(Xiǎohuóluò-dān) on Freund's Complete Adjuvant Induced Arthritic Rats)

  • 윤순모;정만진;권오곤;우창훈;안희덕
    • 한방재활의학과학회지
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    • 제21권1호
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    • pp.97-114
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    • 2011
  • Objectives : The object of this study is to observe the favorable anti arthritic effects of Sowhalrack-dan($Xi\check{a}ohu\acute{o}lu\grave{o}-d\bar{a}n$)(SWRD), which has been traditionally used in Korean medicine to treat rheumatoid arthritis on Freund's complete adjuvant(FCA) induced arthritic Wistar rats. Methods : Rheumatoid arthritis was induced by intradermal injection of FCA(10 mg in 1 ml paraffin oil 0.1 ml/rats). Each of 8 rats showing regular ankle circumferences per group were selected in 14 days after FCA treatment to confirm the induction of rheumatoid arthritis. 300, 150 or 75 mg/kg of SWRD was orally administered once a day for 14 days from 14 days after FCA treatments. Dexamethasone was intraperitoneally administered 15 mg/kg, once a day for 14 days from 14 days after FCA treatments. Rats were sacrificed after 14 days of continuous oral treatment of SWRD or intraperitoneal administration of dexamethasone, and changes were observed; the body weight, knee circumferences, gross arthritis score, inflammatory tissue $prostaglandin(PG)E_2$ levels and cartilage collagen, glucosaminoglycans compositions - chondroitin sulphate, heparin sulphate and hyaluronic acid in the present study. Results : As results of FCA treatment, classic rheumatoid arthritis featuring dramatical decreases on the body weights, cartilage collagen contents and bone glucosaminoglycans - chondroitin sulphate, heparin sulphate and hyaluronic acid contents. Also, it increases the knee circumferences, gross arthritis scores and inflammatory tissue $PGE_2$ levels. However, these changes from FCA induced rheumatoid arthritis were clearly reduced due to the dexamethasone and both two different dosages of SWRD, 300 and 150 mg/kg in the present study. Although FCA induced arthritis were more favorably inhibited by treatment of dexamethasone 15 mg/kg compared to SWRD 300 mg/kg, marked decreases of body weights were detected in dexamethasone 15 mg/kg treated rats. Conclusions : The results obtained in this study suggest that over 150 mg/kg of SWRD showed favorable anti-arthritic effects on the FCA induced arthritis mediated by suppression of $PGE_2$. However, detailed mechanism studies are needed with the screening of the biological active compounds in SWRD. Although FCA induced arthritis were more favorably inhibited by treatment of dexamethasone 15 mg/kg compared to SWRD 300 mg/kg, marked decreases of body weights were detected in dexamethasone 15 mg/kg treated rats, in the present study.

Energy metabolism and whole-exome sequencing-based analysis of Sasang constitution: a pilot study

  • Kim, Hyoung Kyu;Lee, Heetak;So, Ji Ho;Jeong, Seung Hun;Seo, Dae Yun;Kim, Jong-Yeol;Kim, Sanguk;Han, Jin
    • Integrative Medicine Research
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    • 제6권2호
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    • pp.165-178
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    • 2017
  • Background: Traditional Korean Sasang constitutional (SC) medicine categorizes individuals into four constitutional types [Tae-eum (TE), So-eum (SE), Tae-yang (TY), or So-yang (SY)] based on biological and physiological characteristics. As these characteristics are closely related to the bioenergetics of the human body, we assessed the correlation between SC type and energy metabolism features. Methods: Forty healthy, young ($22.3{\pm}1.4$ years) males volunteered to participate in this study. Participants answered an SC questionnaire, and their face shape, voice tone, and body shape were assessed using an SC analysis tool. Thirty-one participants (10 TE, 10 SE, 3 TY, and 8 SY) were selected for further analysis. Collected blood samples were subjected to blood composition analysis, mitochondrial function analysis, and whole-exome sequencing. Results: The SY type showed significantly lower total cholesterol and high-density lipoprotein cholesterol levels than the SE type. Cellular and mitochondrial Adenosine triphosphate (ATP) levels were similar across types. All types showed similar basal mitochondrial oxygen consumption rates, whereas the TE type showed a significantly lower ATP-linked oxygen consumption rate than the other types. Whole-exome sequencing identified several genes variants that were exclusively detected in particular SC types, including 19 for SE, seven for SY, 11 for TE, and six for TY. Conclusion: SC type-specific differences in mitochondrial function and gene mutations were detected in a small group of healthy, young Korean males. These results are expected to greatly improve the accurate screening and utilization of SC medicine.

Screening of key miRNAs related with the differentiation of subcutaneous adipocytes and the validation of miR-133a-3p functional significance in goats

  • Xin, Li;Hao, Zhang;Yong, Wang;Yanyan, Li;Youli, Wang;Jiangjiang, Zhu;Yaqiu, Lin
    • Animal Bioscience
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    • 제36권1호
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    • pp.144-155
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    • 2023
  • Objective: Adipocyte differentiation is regulated by a variety of functional genes and noncoding RNAs. However, the role of miRNAs in lipid deposition of goat white adipose tissue is still unclear. Therefore, this study revealed the miRNA expression profile in goat subcutaneous adipocytes by sRNA-seq. Methods: The miRNA expressed in goat subcutaneous preadipocytes and the mature adipocytes were sequenced by sRNA-seq. The differentially expressed miRNAs (DEm) were screened and gene ontology (GO) and Kyoto encyclopedia for genes and genomes (KEGG) analyses were performed. Gain-of-function and loss-of-function combined with oil red O staining, Bodipy staining, and quantitative reverse-transcription polymerase chain reaction (qPCR) were utilized to determine the effect of miR-133a-3p on adipocyte differentiation. Results: A total of 218 DEm were screened out. The target genes of these DEm were significantly enriched in GO items such as biological regulation and in KEGG terms such as FAK signaling pathway and MAPK signaling pathway. qPCR verified that the expression trend of miRNA was consistent with miRNA-seq. The gain-of-function or loss-of-function of miR-133a-3p showed that it promoted or inhibited the accumulation of lipid droplets, and CCAAT enhancer binding protein α (C/EBPα) and C/EBPβ were extremely significantly up-regulated or down-regulated respectively (p<0.01), the loss-of-function also led to a significant down-regulation of peroxisome proliferator activated receptor gamma (PPARγ) (p<0.01). Conclusion: This study successfully identified miRNAs expression patterns in goat subcutaneous adipocytes, and functional identification indicates that miR-133a-3p is a positive regulator of the differentiation process of goat subcutaneous adipocytes. Our results lay the foundation for the molecular mechanism of lipid deposition in meat-source goats from the perspective of miRNA.

Enhanced Drug Carriage Efficiency of Curcumin-Loaded PLGA Nanoparticles in Combating Diabetic Nephropathy via Mitigation of Renal Apoptosis

  • Asmita Samadder;Banani Bhattacharjee;Sudatta Dey;Arnob Chakrovorty;Rishita Dey;Priyanka Sow;Debojyoti Tarafdar;Maharaj Biswas;Sisir Nandi
    • 대한약침학회지
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    • 제27권1호
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    • pp.1-13
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    • 2024
  • Background: Diabetic nephropathy (DN) is one of the major complications of chronic hyperglycaemia affecting normal kidney functioning. The ayurvedic medicine curcumin (CUR) is pharmaceutically accepted for its vast biological effects. Objectives: The Curcuma-derived diferuloylmethane compound CUR, loaded on Poly (lactide-co-glycolic) acid (PLGA) nanoparticles was utilized to combat DN-induced renal apoptosis by selectively targeting and modulating Bcl2. Methods: Upon in silico molecular docking and screening study CUR was selected as the core phytocompound for nanoparticle formulation. PLGA-nano-encapsulated-curcumin (NCUR) were synthesized following standard solvent displacement method. The NCUR were characterized for shape, size and other physico-chemical properties by Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Fourier-Transform Infrared (FTIR) Spectroscopy studies. For in vivo validation of nephro-protective effects, Mus musculus were pre-treated with CUR at a dose of 50 mg/kg b.w. and NCUR at a dose of 25 mg/kg b.w. (dose 1), 12.5 mg/kg b.w (dose 2) followed by alloxan administration (100 mg/kg b.w) and serum glucose levels, histopathology and immunofluorescence study were conducted. Results: The in silico study revealed a strong affinity of CUR towards Bcl2 (dock score -10.94 Kcal/mol). The synthesized NCUR were of even shape, devoid of cracks and holes with mean size of ~80 nm having -7.53 mV zeta potential. Dose 1 efficiently improved serum glucose levels, tissue-specific expression of Bcl2 and reduced glomerular space and glomerular sclerosis in comparison to hyperglycaemic group. Conclusion: This study essentially validates the potential of NCUR to inhibit DN by reducing blood glucose level and mitigating glomerular apoptosis by selectively promoting Bcl2 protein expression in kidney tissue.

Structural analysis, anti-inflammatory activity of the main water-soluble acidic polysaccharides (AGBP-A3) from Panax quinquefolius L berry

  • Zhihao Zhang;Huijiao Yan;Hidayat Hussain;Xiangfeng Chen;Jeong Hill Park;Sung Won Kwon;Lei Xie;Bowen Zheng;Xiaohui Xu;Daijie Wang;Jinao Duan
    • Journal of Ginseng Research
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    • 제48권5호
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    • pp.454-463
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    • 2024
  • Background: Panax quinquefolius L, widely recognized for its valuable contributions to medicine, has aroused considerable attention globally. Different from the extensive research has been dedicated to the root of P. quinquefolius, its berry has received relatively scant focus. Given its promising medicinal properties, this study was focused on the structural characterizations and anti-inflammatory potential of acidic polysaccharides from the P. quinquefolius berry. Materials and methods: P. quinquefolius berry was extracted with hot water, precipitated by alcohol, separated by DEAE-52-cellulose column to give a series of fractions. One of these fractions was further purified via Sephadex G-200 column to give three fractions. Then, the main fraction named as AGBP-A3 was characterized by methylation analysis, NMR spectroscopy, etc. Its anti-inflammatory activity was assessed by RAW 264.7 cell model, zebrafish model and molecular docking. Results: The main chain comprised of α-L-Rhap, α-D-GalAp and β-D-Galp, while the branch consisted mainly of α-L-Araf, β-D-Glcp, α-D-GalAp, β-D-Galp. The RAW264.7 cell assay results showed that the inhibition rates against IL-6 and IL-1β secretion at the concentration of 625 ng/mL were 24.83 %, 11.84 %, while the inhibition rate against IL-10 secretion was 70.17 % at the concentration of 312 ng/mL. In the zebrafish assay, the migrating neutrophils were significantly reduced in number, and their migration to inflammatory tissues was inhibited. Molecular docking predictions correlated well with the results of the anti-inflammatory assay. Conclusion: The present study demonstrated the structure of acidic polysaccharides of P. quinquefolius berry and their effect on inflammation, providing a reference for screening anti-inflammatory drugs.

가미지패산(加味芷貝散)의 포도상구균 감염 유방염에 대한 항균활성 및 항염 효과 (Effect of Gamijipaesan Extracts against Mastitis Induced by Staphylococcus aureus Infection in a Rat Model through Anti-inflammatory and Antibacterial Effects)

  • 권지명;김동철
    • 대한한방부인과학회지
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    • 제26권1호
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    • pp.1-24
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    • 2013
  • Objectives: The object of this study was to observe the protective effect of Gamijipaesan aqueous extracts(GJS), which has been traditionally used in Korean medicine in obstetrics & gynecological fields as anti-infectious and anti-inflammatory agents, against mastitis induced by Staphylococcus aureus infection in a rat model through antibacterial, antiinflammatory, immunomodulatory, and anti-oxidant effects. Methods: Antibacterial activities of GJS against S. aureus were detected using standard agar microdilution methods, with the effects on the bacterial invasion and intracellular killing of individual test materials in human mammary gland carcinoma cell(MCF-7) and murine macrophages(Raw 264.7) at MIC1/2, MIC and MIC2 concentration levels. In addition, the effects on the cell viability, nitric oxide(NO), tumor necrosis factor(TNF)-${\alpha}$ and interleukin (IL)-6 productions of LPS activated Raw 264.7 cells. The changes on the mammary tissue viable bacterial numbers, myeloperoxidae(MPO), inducible nitric oxide synthetase(iNOS), TNF-${\alpha}$ and IL-6 contents were observed in the S. aureus in vivo intramammary infectious rat model. The anti-bacterial and anti-inflammatory effects were compared with ciprofloxacin and piroxicam, respectively in the present study. Results: MIC of GJS and ciprofloxacin against S. aureus were detected as $0.860{\pm}0.428$ (0.391-1.563) mg/ml and $0.371{\pm}0.262$(0.098-0.782) ${\mu}g/ml$, respectively. In addition, GJS and ciprofloxacin were also showed marked dosage-dependent inhibition of the both bacterial invasion and intracellular killing assays using MCF-7 and Raw 264.7 cells at MIC1/2, MIC and $MIC{\times}2$ concentrations, respectively. $ED_{50}$ against LPS-induced cell viabilities and NO, TNF-${\alpha}$ and IL-6 releases of GJS were detected as 0.72, 0.04, 0.08 and 0.11 mg/ml, and as 19.04, 4.18, 5.37 and 4.27 ${\mu}g/ml$ in piroxicam, respectively. 250 and 500 mg/kg of GJS also inhibit the intramammary bacterial growth, MPO, iNOS, TNF-${\alpha}$ and IL-6 contents in S. aureus in vivo intramammary infected rats, respectively. GJS 500 mg/kg showed quite similar antibacterial and anti-infectious effects as compared with ciprofloxacin 40 mg/kg and also showed similar anti-inflammatory effects as piroxicam 10 mg/kg, in S. aureus in vivo intramammary infectious models. Conclusions: The results obtained in this study suggest that over 250 mg/kg of GJS showed favorable anti-infectious effects against S. aureus infection in a rat model through their antibacterial, anti-inflammatory, immunomodulatory and anti-oxidant effects and therefore expected that GJS can be used as alternative therapies, having both anti-inflammatory and anti-infectious activities. However, more detail mechanism studies should be conducted in future with the efficacy tests of individual herbal composition of GJS and the screening of the biological active compounds in individual herbs. In the present study, GJS 500 mg/kg showed quite similar anti-infectious effects were detected as compared with ciprofloxacin 40 mg/kg treated rats, and also GJS shows quite similar anti-inflammatory effects as compared with piroxicam 10 mg/kg in S. aureus in vivo intramammary infectious rats, but ciprofloxacin did not showed any anti-inflammatory effects, and piroxicam did not showed anti-infectious effects in this study.

청상방풍탕 열수 추출물의 피부재생, 주름개선, 미백, 보습 효과 및 세포독성 평가 (Skin Regeneration, Anti-wrinkle, Whitening and Moisturizing Effects of Cheongsangbangpung-tang Aqueous Extracts with Cytotoxicity)

  • 우창윤;김동철
    • 대한한방부인과학회지
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    • 제30권2호
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    • pp.49-70
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    • 2017
  • 목 적: 본 연구는 한의학에서 면포질환에 빈번히 사용하는 대표적인 복합처방인 청상방풍탕의 열수 추출물 동결건조물의 피부 노화 개선 효과 평가를 위하여 피부재생, 주름개선, 미백 및 보습 효과를 각각 평가하였고, 기본적인 독성 평가의 일환으로 세포독성 또한 in vitro 방법으로 평가하였다. 방 법: 본 연구에서는 human normal fibroblast(CRL-2076) 세포 및 B16/F10 murine melanoma(CRL-6475) 세포에 대한 청상방풍탕 열수 추출 동결건조물(수율 18.71%)의 세포독성을 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) 방법을 통해 평가하였고, B16/F10 melanoma cells의 melanin 생성 억제 정도 및 melanin 합성 필수 효소인 tyrosinase 활성 억제와 arbutin을 비교 평가하여 미백효과를 평가하였다. 또한 피부 재생 및 주름 개선 효과를 transforming growth factor(TGF)-${\beta}1$와 fibroblast의 collagen type I 합성능을 비교, phosphoramidon disodium salt(PP)와 세포외 기질의 분해에 관여하는 elastase활성 억제를 비교, oleanolic acid(OA)와 hyaluronidase, collagenase 및 matrix metalloproteinase(MMP)-1 활성 억제를 비교하여 각각 평가하였고, 마우스 피부 수분함량 변화를 관찰하여 보습효과를 평가하였다. 결 과: 본 실험의 결과, 청상방풍탕은 human normal fibroblast 세포 및 B16/F10 murine melanoma 세포에서 최고 농도인 500 mg/ml까지 유의한 세포독성이 나타나지 않았고, fibroblast의 collagen type I 합성을 증가시켰다. 또한 세포외기질 파괴와 연관된 것으로 알려진 hyaluronidase, elastase, collagenase 및 MMP-1 활성을 각각 억제하였고 melanin의 생성에 관여하는 tyrosinase의 활성 및 B16/F10 melanoma 세포의 melanin 생성을 농도 의존적으로 차단함이 관찰되었다. 이와 더불어 정상매체 대조군에 비해 청상방풍탕 경구 투여군이 투여 용량 의존적으로 마우스 피부 수분 함량을 유의성 있게 증가시켰다. 결 론: 이상의 결과에서, 청상방풍탕은 세포 독성 없이 피부재생, 주름개선, 미백 및 보습 효과를 나타남이 관찰되어 차후 피부 개선 소재로서 그 가치가 높을 것으로 판단되나 금후 개별 구성 약재 각각에 대한 효능 및 생리활성을 나타내는 화학성분의 검색과 더불어 다양한 방면으로 기전적인 연구와 피부 보호 효과에 대한 in vivo평가를 체계적으로 수행해야 할 것으로 판단된다.