• Title/Summary/Keyword: human keratinocyte

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Physiological Activity and Physicochemical Properties of Condensed Prunus mume Juice Prepared with Pectinase (Pectinase처리를 한 매실 농축액의 이화학적 특성 및 생리활성)

  • Kim, Jeong-Ho;Cho, Hyun-Dong;Won, Yeong-Seon;Park, Wool-Lim;Lee, Kwan-Woo;Kim, Hyuk-Joo;Seo, Kwon-Il
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1369-1378
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    • 2018
  • Prunus mume Siebold & Zucc., a member of the Rosaceae family (called Maesil in Korea), has been widely distributed in East Asia, e.g. Korea, Japan and China, and its fruit has been used as a traditional drug and health food. In this study, we evaluated physicochemical properties and physiological activities of condensed Prunus mume juice treated with pectinase (PJ). The values of total acidity, pH, sugar contents, turbidity moisture content of the PJ were 35.81%, 2.73, $54.36^{\circ}Brix$, 2.75 and 51.32%, respectively. The PJ had effective DPPH radical scavenging activity, reducing power effect, $H_2O_2$ scavenging activity and ${\beta}$-carotene bleaching effect. DPPH radical scavenging activities of PJ was 46.31%; their reducing power ($OD_{700}$) was 1.80; $H_2O_2$ scavenging activity of PJ was 91.62%; and ${\beta}$-carotene bleaching effect of PJ was 73.02%. Also, PJ showed effective levels of ${\alpha}$-glucosidase inhibition activity. The cell viability was measured by SRB assay. The PJ significantly decreased the cell viability of mouse melanoma cells (B16) and human melanoma cells (SK-MEL-2 and SK-MEL-28) in a dose-dependent manner, however, there was no effect on human keratinocyte HaCaT. In morphological study, PJ-treated SK-MEL-2 cells showed distorted and shrunken cell masses. Total polyphenol contents and total flavonoid contents of PJ were 588.31 mg% (gallic acid equivalent) and 860.45 mg% (rutin equivalent). The antiproliferative effect of PJ seems to be associated with the antioxidant activity of its flavonoid and polyphenol contents. In conclusion, PJ may be beneficial in development of a functional food material.

Atorvastatin and Fluvastatin Can Reduce IL-1β-induced Inflammatory Responses in Human Keratinocytes (Atorvastatin 그리고 fluvastatin 약물의 IL-1β-유도 염증반응 억제 효과)

  • Choe, Yeong-In;Moon, Kyoung Mi;Yoo, Jae Cheal;Byun, June-Ho;Hwang, Sun-Chul;Moon, Dong Kyu;Woo, Dong Kyun
    • Journal of Life Science
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    • v.31 no.4
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    • pp.418-424
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    • 2021
  • Skin inflammation (dermatitis) is caused by varying skin damage due to ultraviolet radiation and microbial infection. Currently prescribed drugs for dermatitis include anti-histamine and steroid drug classes that soothe inflammation. However, incorrect or prolonged use of steroids can cause weakening of skin barriers as well as osteoporosis. Therefore, treating dermatitis with a drug that has minimal side effects is important. Statins, also known as 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors, are cholesterol-lowering drugs that have been widely treated for hyperlipidemia and cardiovascular diseases. Interestingly, recent studies have shown the anti-inflammatory effects of statins in both experimental and clinical models for of osteoarthritis. This study investigated the possible anti-inflammatory effects of atorvastatin and fluvastatin in human keratinocytes (HaCaT cells), which are crucial components of skin barriers. Stimulation of HaCaT cells with IL-1β increased the expression of the COX2 protein, a major player of inflammatory responses. However, this induction of the COX2 protein was downregulated by pretreatments with atorvastatin and fluvastatin. Treatment with IL-1ß-induced the upregulation of other inflammatory genes (such as iNOS and MMP-1) and these expressions were similarly lowered by these two statin drug treatments. Taken together, these results indicated that atorvastatin and fluvastatin can reduce IL-1β-induced inflammatory responses in HaCaT cells. In conclusion, the findings suggest that atorvastatin and fluvastatin can be potential modulators for ameliorating skin inflammation.

Effects of Anti-inflammation and Skin Barrier by Genistein Cyclodextrin Complex (제니스테인 Cyclodextrin 포접체의 항염 및 피부장벽에 미치는 영향)

  • Choi, Dong Jun;Cho, Uk Min;Choi, Da Hee;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.171-181
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    • 2018
  • Genistein is one of the representative isoflavone compounds isolated from soybeans and has been studied very well for its anti-aging and anti-inflammatory activity through previous studies. However, although genistein exhibits high solubility in organic solvents, it shows low bioavaility due to the low water solubility. In this study, we compared directly the functional difference between genistein and genistein cyclodextrin complex which has the improved water solubility and stability by cell based assay. Cell cytotoxicity experiment were carried out on RAW264.7 with CCK-8 assay and cytotoxicity was appeared from $10{\mu}g/mL$, thereby maximum concentration was set to $10{\mu}g/mL$ in all condition. We discovered that genistein CD complex suppressed NO production and iNOS expression as concentration dependent manner in the condition of LPS rather than genistein. Also, we could understand that genistein CD complex was able to down-regulate mRNA expression of anti-inflammatory cytokines such as $IL1-{\alpha}$, $IL1-{\beta}$, IL-6, and $TNF-{\alpha}$ as concentration dependent manner in the presence of LPS. In addition, we verified that genistein CD complex increased TEER of HaCaT human keratinocyte cells as concentration dependent pattern and stimulated cell division and migration rather than genistein in cell migration assay. Thus, it is expected that it can be used as an effective cosmetic raw material for improving atopic dermatitis or skin barrier if clinical studies on skin regeneration and skin barrier of the genistein CD complex are carried out.

Development of Natural Antioxidants and Whitening Agents for Cosmeceuticals

  • Kim, Jong-Pyung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.11a
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    • pp.79-92
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    • 2007
  • Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

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Gene Expression Profiles and Antioxidant Effects of Houttuynia cordata Thunb Extract in Human Keratinocyte HaCaT Cells (인간 피부각질세포 HaCaT에서 어성초 추출물의 유전체 발현 분석 및 항산화 효과)

  • Kim, Jung Min;Bang, In Seok
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1406-1415
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    • 2018
  • Based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of Houttuynia cordata Thunb, the cytoprotective effects by oxidative-stress were here analyzed. Regarding the antioxidant activity of organic solvent fractions, the electron-donating ability of DPPH increased in a dose-dependent manner, and $ED_{50}$ exhibited the highest concentration at $175{\mu}g/ml$ in the Hc-EtOAc fraction. The cell viability of Hc-EtOAc fractions on $H_2O_2$-induced HaCaT cell death ($IC_{50}$) increased in a concentration-dependent manner and a visible cell survival rate of 74% was observed at a concentration of $100{\mu}g/ml$. Meanwhile, the gene expression patterns in HaCaT cells treated with $100{\mu}g/ml$ of the Hc-EtOAc fraction for 6 and 24 hr were identified with microarray analysis. The genes involved in signal transduction, cell division, antioxidant activity, and epithelial cell proliferation were found to be 2-fold up-regulated genes in HaCaT cells following the Hc-EtOAc fraction treatment. Especially, proinflammatory cytokines (IL1B, TNF, and IL6) were identified as involved in antioxidant activity based on the expression patterns of the HaCaT cells, and pathway analysis indicated that TLR4 might be considered an upstream regulator of these genes. In order to verify the activity of IL1B, TNF, and IL6, qRT-PCR showed that the expression increased more than 2 times in HaCaT cells treated with at least $100{\mu}g/ml$ of the Hc-EtOAc fraction. The activity of the upstream regulator TLR4 protein was also increased by the Hc-EtOAc fraction. As a result, the antioxidative activity of the Hc-EtOAc fraction is predicted to pass from TLR4 through cytokines such as IL1B, TNF, and IL6.

Studies on Antioxidant, Anti-inflammatory and Whitening Effects of Oriental Herbal Extracts (Mix) including Eucommiae cortex (두충을 포함하는 한방추출물(Mix)의 항노화, 항염, 미백 효능 활성에 관한 연구)

  • Choi, Da Hee;Kim, Mi Ran;Kim, Min Young;Kim, Ho Hyun;Park, Sun-Young;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.1
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    • pp.37-47
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    • 2019
  • Recently, due to the increase in skin diseases caused by particulate matter, endocrine disruptor and environmental changes, the trend of development of cosmetic materials has been shifting to the more safe and effective ingredients based on natural materials rather than existing synthetic compounds like steroids and antihistamines. This study aimed to develop a new natural cosmetic materials using oriental herbs such as Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus. First, DPPH assay was performed to examine the antioxidative activity of the herbal extract (Mix) and 98.8% DPPH radical scavenging activity was confirmed at $400{\mu}g/mL$ concentration of it. In order to confirm the whitening efficacy of oriental herbal extracts(mix), the amount of melanin synthesized after stimulation of ${\alpha}-MSH$ with B16F10 cells was measured. Results showed that it was decreased to 27.1% comparing with the only ${\alpha}-MSH$ treated group, which confirmed the whitening efficacy. Also, both nitric oxide(NO) production and iNOS and COX-2 expression were significantly reduced in RAW264.7 macrophages activated by LPS in the presence of the extracts(Mix). The mRNA expression of the inflammatory cytokines such as $IL-1{\alpha}$, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ was also analyzed to confirm the inhibition effect of the extracts on inflammation. Finally, to confirm the enhancement of skin barrier function, the expression of claudin 1 gene, a tight junction protein, was observed using human keratinocyte HaCaT cells and increased as concentration dependent manner. From these results, it is concluded that the oriental herbal extracts(Mix) containing Eucommiae cortex, Alpinia oxyphylla Miquel and Bombyx batryticatus is effective for antioxidant, anti-inflammation, skin whitening, and skin barrier and thus could be applied as a new natural cosmetic material.