• Title/Summary/Keyword: collagenase activity

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Inhibitory activities on biological enzymes of extracts from Oplismenus undulatifolius (주름조개풀(Oplismenus undulatifolius) 추출물의 생리활성 효소 억제 효과)

  • Lee, Eun-ho;Kim, Byung-Oh;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.60 no.2
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    • pp.101-108
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    • 2017
  • The phenolic contents which were extracted with water and 70% ethanol from O. undulatifolius were 7.7, 10.1 mg/g, respectively. The 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity of water and ethanol extracts were 78, 82% at $50{\mu}g/mL$ phenolics, respectively. The 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization activity were 92, 76% at $100{\mu}g/mL$ phenolics. Antioxidant protection factor in water and ethanol extracts at $200{\mu}g/mL$ phenolics were 1.51 and 2.08 PF, respectively. Thiobarbituric acid reactive substance were 84% in water extracts and 99% in ethanol extracts at $50{\mu}g/mL$ phenolics, respectively. The inhibition activity on ${\alpha}-Glucosidase$ was 44% in ethanol extracts at $200{\mu}g/mL$ phenolics. The inhibition activity on ${\alpha}-amylase$ was 37-88% in water extracts at $50-200{\mu}g/mL$ phenolics. The tyrosinase inhibition activity as whitening effect were 82% in ethanol extracts. The elastase inhibition activity were 4, 61% in water and ethanol extracts, respectively. The collagenase inhibition activity of antiwrinkle effect showed an excellent wrinkle improvement effect as 39% in water extracts and 67% in ethanol extracts at $200{\mu}g/mL$ phenolics, respectively. The hyaluronidase inhibition activity as anti-inflammation effect of ethanol extracts was confirmed to 46% of inhibition at $200{\mu}g/mL$ phenolic. The astringent effect of water and ethanol extracts was confirmed to 13, 32% of effect at $200{\mu}g/mL$ phenolic, respectively.

Mechanisms of Suppression of Matrix Metalloproteinases in UVB-Irradiated HaCaT Keratinocytes of Colored Rice Varieties (UVB에 조사된 HaCaT Keratinocytes에서의 유색미에 의한 Matrix Metalloproteinases 발현억제 메커니즘)

  • Choi, Eun-Young;Lee, Jae-Bong;Kim, Do-Hoon;Kwon, Yong-Sham;Cheon, Jung-Yoon;Lee, Jin-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.562-571
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    • 2017
  • In this study, we investigated the anti-oxidant activities [electron-donating ability (EDA), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging ability, and reactive oxygen species (ROS) inhibitory activity], anti-wrinkle activities [collagenase inhibitory activity, suppression and/or phosphorylation of matrix metalloproteinases (MMPs), and mitogen-activated protein (MAP) kinase activity], and mRNA expression levels using reverse transcription-polymerase chain reaction (RT-PCR) assay in ultraviolet (UV) B ray ($50mJ/cm^2$)-irradiated human keratinocyte HaCaT cells. Josaengheugchal, Sinneungheugchal (SE), Shintoheug rice, Heugjinju rice, and Heugseol (HE) among colored rice varieties were reported to have excellent antioxidant properties. In the EDA and ABTS radical scavenging assays, extracts of the five colored rice varieties had scavenging activities of 72% at concentrations higher $50{\mu}g/mL$. In the collagenase inhibition assay, ethanol extracts of the five colored rice varieties showed high inhibitory effects of about 60% at concentrations higher $25{\mu}g/mL$. In the ROS inhibition assay, ethanol extracts of HE and SE showed very excellent inhibition efficacies at all concentrations. We determined molecular biological mechanisms of MMPs (MMP-1, -3, -8, and -13) and mitogen-activated protein kinase (MAPK) with HE, and the results show that HE suppressed expression of MMPs and phosphorylation of MAPK and increased expression of pro-collagen type I in UVB-irradiated cells. It was also confirmed by RT-PCR that HE reduced expression of MMPs mRNA. Therefore, these results suggest that HE has anti-wrinkle and collagen production effects and may be used as a material in the development of functional food and cosmetic industries.

Thiol-dependent Redox Mechanisms in the Modification of ATP-Sensitive Potassium Channels in Rabbit Ventricular Myocytes

  • Han, Jin;Kim, Na-Ri;Cuong, Dang-Van;Kim, Chung-Hui;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.15-23
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    • 2003
  • Cellular redox state is known to be perturbed during ischemia and that $Ca^{2+}$ and $K^2$ channels have been shown to have functional thiol groups. In this study, the properties of thiol redox modulation of the ATP-sensitive $K^2$ ($K_{ATP}$) channel were examined in rabbit ventricular myocytes. Rabbit ventricular myocytes were isolated using a Langendorff column for coronary perfusion and collagenase. Single-channel currents were measured in excised membrane patch configuration of patch-clamp technique. The thiol oxidizing agent 5,5'-dithio-bis-(2-nitro-benzoic acid) (DTNB) inhibited the channel activity, and the inhibitory effect of DTNB was reversed by dithiothreitol (disulfide reducing agent; DTT). DTT itself did not have any effect on the channel activity. However, in the patches excised from the metabolically compromised cells, DTT increased the channel activity. DTT had no effect on the inhibitory action by ATP, showing that thiol oxidation was not involved in the blocking mechanism of ATP. There were no statistical difference in the single channel conductance for the oxidized and reduced states of the channel. Analysis of the open and closed time distributions showed that DTNB had no effect on open and closed time distributions shorter than 4 ms. On the other hand, DTNB decreased the life time of bursts and increased the interburst interval. N-ethylmaleimide (NEM), a substance that reacts with thiol groups of cystein residues in proteins, induced irreversible closure of the channel. The thiol oxidizing agents (DTNB, NEM) inhibited of the $K_{ATP}$ channel only, when added to the cytoplasmic side. The results suggested that metabolism-induced changes in the thiol redox can also modulate $K_{ATP}$ channel activity and that a modulatory site of thiol redox may be located on the cytoplasmic side of the $K_{ATP}$ channel in rabbit ventricular myocytes.

Studies on Cosmeceutical Activity of Extracts of Moringa oleifera Extract (모링가 추출물에 대한 화장품약리활성 검증)

  • Kim, So Ra;Yoo, Dan Hee;Yeom, Hyeon Ji;Oh, Min Jeong;Lee, Jin Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.219-229
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    • 2018
  • The purpose of this study was to investigate the role of the Moringa oleifera (M. oleifera) extract as a cosmetic additive. The tyrosinase and elastase inhibitory effects showed 47% and 39% at $1,000{\mu}g/mL$ concentration, respectively. Also, the collagenase inhibition effect was 31% at $500{\mu}g/mL$ concentration. A cell viability test, measured on macrophage cell (RAW 264.7) and melanoma cell (B16F10) by ethanol extract of M. oleifera, showed 94.2% and 94.8% at $100{\mu}g/mL$ concentration, respectively. In order to confirm anti-inflammatory activity, we examined the inhibitory effects on the production of lipopolysaccharides (LPS)-induced NO in RAW 264.7 cells by Griess assay. As a result, the M. oleifera extract showed a concentration-dependent inhibition of NO production. The protein expression inhibitory effects of M. oleifera extract were measured by western blot at 25, 50, $100{\mu}g/mL$ concentration and the ${\beta}-actin$. Results showed that the expression inhibition rates of the iNOS, COX-2, MITF, TRP-1, TRP-2, tyrosinase protein were decreased by 85.8%, 57.5%, 80.7%, 30%, 29.9%, 23.6% at $100{\mu}g/mL$ concentration, respectively. It was concluded that M. oleifera extracts had the anti-inflammatory and whitening effects and thus could be applied for cosmetics as a natural ingredient.

Inhibitory Efficacy of Smilax china L. on Pro-collagen Type-1 Activity and MMP-1 Gene Expression in Fibroblasts (CCD-986sk) (섬유아세포(CCD-986sk)에서 청미래덩굴의 pro-collagen type-1 활성 및 MMP-1의 유전자 발현 저해능)

  • Lee, Soo-Yeon;Lee, Jin-Young
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1239-1245
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    • 2013
  • The purpose of this study was to research the biological activity of ethanol extract from Smilax china L. which is a vine shrub belonging to the lily family. For antiwrinkle effects, elastase inhibition effect of ethanol and water extracts from S. china L. showed 41.1% and 16.3% at $1,000{\mu}g/ml$ concentration. The collagenase inhibition effect of ethanol and water extracts from S. china L. showed more than 96.6% and 60.0% at $1,000{\mu}g/ml$ concentration. As a result of having fibroblast measured cell viability on fibroblast cell of ethanol extract from S. china L., it showed 71.7% with cell viability at $100{\mu}g/ml$ concentration. At $50{\mu}g/ml$ concentration, the procollagen biosynthesis effect of ethanol extract from S. china L. was 139.86%. At the same concentration, the matrix metalloprotease (MMP)-1 inhibition effect of the ethanol extract was 74.9%. According to the results of Western blot of ethanol extract from S. china L., the expression of the MMP-1 protein was decreased by 35% at $50{\mu}g/ml$ concentration. Reverse transcription-polymerase chain reaction (PCR) of ethanol extract from S. china L. showed that the expression of MMP-1 mRNA was decreased by 45% at $50{\mu}g/ml$ concentration. The findings suggest that 70% ethanol extract from S. china L. (SC) has great potential as a cosmeceutical ingredient with antiwrinkle effects.

THE STUDY ON THE EFFECTS OF THE INSULIN-LIKE GROWTH FACTOR-I ON THE BIOLOGICAL ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Insulin-like growth factor-I 이 치주인대세포의 생물학적 활성도에 미치는 영향에 대한 연구)

  • Kim, Seong-Jin;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.219-237
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    • 1994
  • The ultimate goal of clinical periodontal therapy is to achieve regeneration of a healthy connective tissue reattachment. Conventional therapy including scaling, root planing, gingival curettage, gingivectomy and flap procedures of various types results primarily in repair rather than regeneration of the periodontium. In order for periodontal regeneration to occur, progenitor periodontal ligament cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. Insulin-like growth factor-I (IGF- I ) of these factors appear to have an important role in periodontal wound healing and bone formation. The purpose of this study is to evaluate the effects of IGF- I on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were obtained from periodontal tissue explants culture of the first premolar tooth extracted for the orthodontic treatment. Cells were cultured in Dulbecco's modified Eagle medium(DMEM) with 10% fetal bovine serum. Fourth to seventh passage cells were plated in 24 well tissue culture plates and medium changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of $[^3H]-thymidine$ into DNA, Protein synthesis was determined by measurement of $[^3H]-proline$ incorporation into collagenase-digestible protein(CDP) and noncollagenous protein(NCP) according to the method of Peterkofsky and Diegelmann (1971), And alkaline phosphatase activity was measured as one parameter of osteoblastic differentiation. The results were as follows : The DNA synthetic activity was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I At the concentration of 10, 100ng/ml, IGF- I significantly increased the DNA synthetic activity(P<0.05) The total protein, collagen and noncollagen synthesis was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I. At the concentration of 1, 10, 100ng/ml, IGF- I significantly increased the total protein, collagen and noncollagen synthesis activity(P<0.95, P<0.001). The % of collagen was not effected according to the concentration of IGF- I. The alkaline phosphatase activity was increased in a dose-, time-dependent manner with IGF- I (10, 100ng/ml). In conclusions, the present study shows that IGF- I has a potentiality to enhance the DNA synthesis of periodontal ligament cells with including the increase of the total protein and collagen synthetic activity. The use of IGF- I to mediate biological stimulation of periodontal ligament cells shows promise for future therapeutic applications.

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Whitening and anti-wrinkle effect of Pinus koraiensis leaves extracts according to the drying technique (건조방법에 따른 잣나무(Pinus koraiensis) 잎 추출물의 주름개선 및 미백 효과)

  • Jo, Jae-Bum;Park, Hye-Jin;Lee, Eun-Ho;Lee, Jae-Eun;Lim, Su-Bin;Hong, Shin-Hyub;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.60 no.1
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    • pp.73-78
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    • 2017
  • This study provide activity for beauty food of water and 80 % ethanol extracts from Pinus koraiensis leaves. Total phenolic content of extracts from Pinus koraiensis leaves were each 12.22 mg/g (Drying under hot air) and 17.93 mg/g (Drying under shade), 14.36 mg/g (Lyophilization) in water extracts (WE) and 11.9 mg/g and 20.63 mg/g, 17.96 mg/g in 80 % ethanol extracts (EE). The 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity of extracts from Pinus koraiensis leaves was 96.20 % in EE from drying under shade at extracts concentration. The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical decolorization activity of extracts from drying under shade was 99.85 % in WE and 99.80 % in EE at extracts concentration. The antioxidant protection factor (PF) extracts from drying under shade type was 9.63 PF in WE and 10.48 PF in EE at extracts concentration. The thiobarbituric acid reactive substance from Pinus koraiensis leaf was 89.39 % in EE from drying under shade at extracts concentration. The elastase inhibition activity of EE for anti-wrinkle effect showed an excellent wrinkle improvement effect, showing 71.46 % in EE from lyophilization. Collagenase inhibition activity of EE from drying under shade was 97.48 % in extracts. Tyrosinase inhibition activity which was related to anti-melanogensis was observed. The tyrosinase inhibitory effect of extracts from lyophilization was confirmed to be 60.4 % in EE more than another drying methods at extracts concentration. Through out all results, it can be expected Pinus koraiensis leaves extracts to use as a functional material for anti-oxidant and functional beauty food.

Antioxidative and biological activity of extracts from Orostachys japonicus (와송(Orostachys japonicus) 추출물의 항산화 및 생리 활성)

  • Lim, Sun-Mi;Park, Hye-Jin;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.60 no.4
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    • pp.293-300
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    • 2017
  • This study was designed to extracts from Orostachys japonicas were investigated to assess anti-oxidation and biological activity. Phenolic content was maximum of $10.56{\pm}0.32mg/g$ when extracted with 50% ethanol. In anti-oxidative activity, Orostachys japonicus electric donating activity was higher than 80% in both water and ethanol extract at $200{\mu}g/mL$. 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization of both water and ethanol extract was higher than 95.0% but antioxidant protection factor of water extract was higher than ethanol extract. Thiobarbituric acid reactive substance of ethanol extract was higher than water extract. For antihypertensive effect determination, angiotesin converting enzyme of water and ethanol extract showed 6.67 and 7.98% each at $200{\mu}g/mL$. Ethanol extract of $200{\mu}g/mL$ showed xanthin oxidase inhibitory effect of 60.85% but was not shown with water extract. Orostachys japonicus ethanol extract showed higher tyrosinase inhibitory activity of 64.59% which was higher than kojic acid of control indicating higher whitening effect. In anti-wrinkle effect, ethanol extract at $50-200{\mu}g/mL$ showed collagenase inhibitory effect of 75.95-85.02% which was higher than 68.91-76.64% of epigallocatechin-gallate of control group. 50% ethanol extract showed higher elastase inhibitory activity than water extract. Therefore, Orostachys japonicus extracts were identified to have high anti-wrinkle effect. These results identify anti-oxidative activity, gout prevention, whitening effect, and anti-wrinkle effect which indicate the possibility as a source for functional material.

Antioxidant and anti-aging activities of water extracts from Chionanthus retusus flesh according to different extraction temperatures (추출온도에 따른 이팝나무 과육 물 추출물의 항산화 및 항노화 활성)

  • Choi, Hae-Sim;Lee, Yang-Suk;Seo, Su-Jeong;Kim, Nam-Woo
    • Food Science and Preservation
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    • v.24 no.8
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    • pp.1129-1137
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    • 2017
  • The fringe tree, Chionanthus retusus (Oleaceae) has been used as a natural remedy that helps a paralytic stroke, dementia, phlegm and malaria. This study was conducted to investigate the antioxidant and anti-aging activities of water extracts obtained from the flesh of C. retusus by reflux extraction at room temperature and $80^{\circ}C$, and pressure heating extraction at $110^{\circ}C$, for an increment of their practical use. The highest extraction yield from the flesh of C. retusus was 67.94% extract by pressure heating extraction at $110^{\circ}C$. The content of total polyphenol compounds was the highest by 72.71 mg/g in $80^{\circ}C$ extract by reflux extraction. The highest content of total flavonoid compounds was 7.60 mg/g at $110^{\circ}C$ extract. The results of soluble protein and reducing sugar contents showed the highest content of 4.93 mg/g and 46.77 mg/g in $110^{\circ}C$ extract, respectively. In the analysis of DPPH radical scavenging activity, all extracts showed an excellent effect of 92% in 1.0 mg/mL concentration. The highest effect of ABTS radical scavenging activity was 88.67% at $110^{\circ}C$ extraction in 1.0 mg/mL concentration. Nitrite scavenging ability was the highest as 66.16% at $110^{\circ}C$ extract at pH 1.2 and 1.0 mg/mL concentration. The results of SOD-like activity and xanthine oxidase inhibition activities showed the highest effect of 14.95% and 59.45% at $110^{\circ}C$ extract in 1.0 mg/mL, respectively. Tyrosinase inhibition activity for whitening effect was the highest of 35.31% at $85^{\circ}C$ in 1.0 mg/mL. In the analysis of anti-aging effect, collagenase and elastase inhibition activity showed the highest effects of 37.78% at $110^{\circ}C$ and 20.39% at $80^{\circ}C$, respectively. All results of antioxidant activities and anti-aging effects increased along with the concentration increases. These results indicated that the extracts extracted from the flesh of C. retusus at $80^{\circ}C$ and $110^{\circ}C$ have a large amount of useful ingredients, an excellent antioxidant activity, like as DPPH and ABTS radical scavenging activity, and anti-aging effects to develop functional products than those of $25^{\circ}C$.

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

  • Woo, Chang-Yoon;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.2
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    • pp.49-70
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    • 2017
  • Objectives: The present study is to observe the skin-regeneration, anti-wrinkle, whitening and skin moisturizing effects of Cheongsangbangpung-tang (CSBPT) with cytotoxicity. Methods: In the present study, cytotoxicity of CSBPT lyophilized aqueous extracts (yield=18.71%) was experimented against human normal fibroblast cells and B16F10 murine melanoma cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide (MTT) assay, and skin regeneration and anti-wrinkle effects were also showed through the assay of collagen type I synthesis by an enzyme immunoassay (EIA) kit as comparing with transforming growth factor (TGF)-${\beta}1$, hyaluronidase, collagenase and matrix metalloproteinase (MMP)-1 inhibitory assays as comparing with oleanolic acid (OA), and elastase inhibitory effects as comparing with phosphoramidon disodium salt (PP). In addition, whitening effects of CSBPT were observed by tyrosinase inhibitory assay and melanin formation test in B16/F10 melanoma cells as comparing with arbutin, and skin moisturizing effects were measured through mouse skin water contents test, respectively. Results: No CSBPT treatment related cytotoxic effects were demonstrated against human normal fibroblast cells and B16/F10 murine melanoma cells. CSBPT concentration-dependent increased collagen type I synthesis at human normal fibroblast cells. It also effectively suspreessed hyaluronidase, collagenase, elastase and MMP-1 activities, which were enzymes that related to declining of ECM and formation of wrinkle. CSBPT supressed B16/F10 melanoma cells's melanin productions with tyrosinase activity, which was an enzyme connected with melanin formation, and dose-dependent and significant increases of skin water contents were detected in CSBPT treated mouse skin as compared with vehicle control skins. Conclusions: CSBPT showed favorable and enough skin regeneration, anti-wrinkle, whitening and skin moisturizing effects at least in a condition of this experiment. However, more detail mechanism and in vivo skin protective efficacy studies should be conducted in future with the screening of the biological active compounds in individual herbs of Cheongsangbangpung-tang.