• Title/Summary/Keyword: human keratinocyte cell line

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The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

Protective Effect of Glycyrrhiza glabra Extract on UV-induced Skin DNA Damage (감초추출물(Glycyrrhiza glabra Extract)의 피부에서의 DNA 손상 방지효과)

  • Shin, Jae Young;Kang, Nae Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.1
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    • pp.33-38
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    • 2022
  • Ultraviolet B (UVB) damages DNA residues in skin keratinocytes. In particular, the formation of cyclobutane pyrimidine dimers (CPD), a pyrimidine residue damage in DNA, is considered a representative indicator of skin photoaging. In this study, we confirmed defensive effect of Glycyrrhiza glabra (G. glabra) extract against UVB induced DNA damage. First of all, we confirmed UVB dependent amount of CPD formation in human keratinocyte cell line. UVB induced CPD was decreased by G. glabra extract by dose dependent manner. In addition, it was confirmed that the expression of mRNA of DNA damage recovery factors was increased by G. glabra extract. Consequently, through this study, it was possible to confirm the DNA protection effect of G. glabra extract in skin keratinocytes.

Evaluation of Biological Activities on the Extractives of Pinaceae (소나무과 일부 수종의 에탄을 추출물에 대한 생물학적 평가)

  • Ahn, Jeung-Youb;Bae, Jong-Hwan
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.121-125
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    • 2005
  • Most of the cells, specially in the skin, free radicals and reactive oxygen species induced aging, accompanying with diseases. The pinaceae family is very familiar plant with Korean spirit. However, those plants were not estimated for natural products in manufactural process. We employed 4 different plants, Pinus koraienis S et Z. Pinus banksiana LAMBERT. Pinus rigida MILL. Pinus densiflora S. et Z. in pinceae family. The ethanol extracts were prepared for three different parts of plant, wood, bark and leaf, respectively. In this study, we carried out free radical and superoxide radical scavenging assay to investigate the anti-oxidative activity The bark and leaf part of plants showed similar anti-oxidant effect. L-ascorbic acid and $\alpha$-tocopherol at $10{\mu}g/mL$. Also we tried to investigate inhibitory effects of elastase activity in in vitro experiments on pinaceae plant extract as a matters for functional cosmetics. Among those plants, the bark of Pinus rigida MILL. and Pinus densiflora S. et Z. showed inhibitory effect. The cell viability was evaluated with MTT assay. The potential relationship was shown between the cell viability and anti-oxidant effect because the anti-oxidant effects were positively correlated wvith the cell growth in MTT assay. As the results in our experiments, we expect the potential activities of pinaceae as a material of functional cosmetics.

In vitro cytotoxicity of self-etching primers (자가 산부식 프라이머의 세포독성에 관한 실험적 연구)

  • Rhee, Chang-Hoon;Kim, In-Ryeon;Kim, Gyoo-Cheon;Kim, Seong-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.36 no.6
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    • pp.422-433
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    • 2006
  • Objective: Several ions and components are released from self-etching primers in the oral cavity. This may cause injury to the periodontal tissues throughout orthodontic treatment. The purpose of this study was to assess the cytotoxicity of self-etching primers to HGF-1, HaCaT, and RHEK cells. Method: Transbond XT Primer (3M Unitek, Monrovia, CA, USA), and self-etching primers, Clearfil SE Bond (Kuraray, Osaka, Japan), Transbond Plus SEP (3M Unitek, Monrovia, CA, USA), and Adper Prompt L-Pop (3M Unitek, Monrovia, CA, USA), were evaluated by MTT assay, and cellular changes were also observed. Results: In all cells after 72 hours with all primers, severe morphological changes such as atrophy and necrosis were observed. In the MTT assay using HGF-1, Clearfil SE Bond, Transbond XT Primer, Transbond Plus SEP, and Adper Prompt L-Pop were lined up in order of ascending cytotoxicity When using HaCaT, Clearfil SE Bond, Adper Prompt L-Pop, Transbond Plus SEP, and Transbond XT Primer were lined up in order of ascending cytotoxicity. When using RHEK, Clearfil SE Bond, Transbond XT Primer, Adper Prompt L-Pop, and Transbond Plus SEP were lined up in order of ascending cytotoxicity. Conclusion: The result of this study shows that care is needed because self-etching primers show cytotoxic properties similar to conventional primers.

Effect of Fucus evanescens Fucoidan on Expression of Matrix Metalloproteinase-1 Promoter, mRNA, Protein and Signal Pathway (Fucus evanescens fucoidan의 matrix metalloproteinase-1 promoter, mRNA, 단백질 발현과 신호전달경로에 미치는 효과)

  • Ku, Mi-Jeong;Jung, Ji-Won;Lee, Myeong-Sook;Cho, Byung-Kyu;Lee, Soon-Rye;Lee, Hye-Sook;Vischuk, Olesya S.;Zvyagintseva, Tatyana N.;Ermakova, Svetlana P.;Lee, Yong-Hwan
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1603-1610
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    • 2010
  • Fucoidans are sulfated fucosylated polymers from the cell wall of brown algae. We assessed the effects of Fucus evanescens fucoidan on ultraviolet-B (UVB)-induced expression of matrix metalloproteinase-1 (MMP-1) protein, mRNA, and promoter, and the phosphorylation of mitogen-activated protein kinases in vitro using an immortalized human keratinocyte cell line. Pretreatment with 10 and $100\;{\mu}g/ml$ fucoidan significantly inhibited UVB-induced MMP-1 protein, mRNA and promoter activity, compared to UVB irradiation alone. Extracellular signal regulated kinase activation was markedly inhibited by treatment with fucoidan, though c-JUN N-terminal kinase activity and p38 activation were only marginally affected by fucoidan. F. evanescens fucoidan may be a potential therapeutic agent for the prevention and treatment of skin photoaging.

In Vitro and in Vivo Wound Healing Properties of Plasma and Serum from Crocodylus siamensis Blood

  • Jangpromma, Nisachon;Preecharram, Sutthidech;Srilert, Thanawan;Maijaroen, Surachai;Mahakunakorn, Pramote;Nualkaew, Natsajee;Daduang, Sakda;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1140-1147
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    • 2016
  • The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

Protective Effects of Trifolium pratense L. Extract against H2O2-induced Oxidative Stress in HaCaT Keratinocytes (인간 피부각질세포에서 Hydrogen peroxide로 유도된 산화적 스트레스에 대한 붉은 토끼풀 추출물의 세포 보호 효과)

  • Mi Song Shin;You Kyeong Lee;Seo Young Choi;Ji Sun Hwang;Parkyong Song;Hyeon Cheal Park;Keun Ki Kim;Hong-Joo Son;Yu-Jin Kim;Kwang Min Lee
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.2
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    • pp.223-232
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    • 2023
  • Oxidative stress plays a significant role in the pathogenesis of various skin conditions, resulting in cellular and tissue damage that can contribute to the development of skin tone unevenness, roughness and wrinkles. In this study, we found that Trifolium pratense L. extract (TE) attenuated oxidative-induced damage in HaCaT cells and elucidated the underlying molecular mechanism. Our finding demonstrated that TE effectively protected HaCaT cells against H2O2-induced cell death by inhibiting caspase-3 activation, downregulating Bax and upregulating Bcl-2, and attenuating the activation of three mitogen-activated protein kinases (MAPKs). Our results suggest that TE has remarkable cytoprotective properties against oxidative damage in HaCaT cells and could serve as a complementary or alternative approach to prevent and treat skin damage.

Effects of 7-MEGATM 500 on Oxidative Stress, Inflammation, and Skin Regeneration in H2O2-Treated Skin Cells

  • Song, In-Bong;Gu, Hyejung;Han, Hye-Ju;Lee, Na-Young;Cha, Ji-Yun;Son, Yeon-Kyong;Kwon, Jungkee
    • Toxicological Research
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    • v.34 no.2
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    • pp.103-110
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    • 2018
  • Environmental stimuli can lead to the excessive accumulation of reactive oxygen species (ROS), which is one of the risk factors for premature skin aging. Here, we investigated the protective effects of $7-MEGA^{TM}$ 500 (50% palmitoleic acid, 7-MEGA) against oxidative stress-induced cellular damage and its underlying therapeutic mechanisms in the HaCaT human skin keratinocyte cell line (HaCaT cells). Our results showed that treatment with 7-MEGA prior to hydrogen peroxide ($H_2O_2$)-induced damage significantly increased the viability of HaCaT cells. 7-MEGA effectively attenuated generation of $H_2O_2$-induced reactive oxygen species (ROS), and inhibited $H_2O_2$-induced inflammatory factors, such as prostaglandin $E_2$ ($PGE_2$), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), and $interleukin-1{\beta}$ ($IL-1{\beta}$). In addition, cells treated with 7-MEGA exhibited significantly decreased expression of matrix metalloproteinase-1 (MMP-1) and increased expression of procollagen type 1 (PCOL1) and Elastin against oxidative stress by $H_2O_2$. Interestingly, these protective activities of 7-MEGA were similar in scope and of a higher magnitude than those seen with 98.5% palmitoleic acid (PA) obtained from Sigma when given at the same concentration (100 nL/mL). According to our data, 7-MEGA is able to protect HaCaT cells from $H_2O_2$-induced damage through inhibiting cellular oxidative stress and inflammation. Moreover, 7-MEGA may affect skin elasticity maintenance and improve skin wrinkles. These findings indicate that 7-MEGA may be useful as a food supplement for skin health.

The Anti-bacterial Effects and Epidermal Permeability Barrier Function of Red Onion Juice Produced in Jeon-Nam province in Korea (전남 함평산 자색양파 추출물의 항균효과와 피부장벽 보호효과)

  • Youn, Dae-Hwan;Shin, Heon-Tae
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.1
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    • pp.43-56
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    • 2012
  • Objective : In this study, we assessed the anti-bacterial effects and epidermal permeability barrier function of red onion juice comparing to yellow onion juice and $Houttuynia$ $cordata$ extract $in$ $vitro$. Methods : 3types of red and yellow onion juice were prepared as antibacterial agent candidates with Houttuynia cordata hot water extract using 4 different bacterial strains ($Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$, $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$) by colony counting method. The expression of filaggrin, a marker of keratinocyte differentiation, and serine palmitoyl transferase (SPT), a marker of the formation of the stratum corneum lipid barrier, in human HaCat keratinocytes were analyzed using HaCaT cell line. The expression of COX-2 and AP-1 which is a factor of COX-2 transcription were also analyzed by western blotting method. Results : There was detectable anti-bacterial effects on $Staphylococcus$ $epidermidis$, $Staphylococcus$ $aureus$ $subsp$ among 1%, 5%, 10% extracts of yellow and red onion.(81%-100%) The bacteriocidal effects were not shown on $Escherichia$ $coil$, $Salmonella$ $enterica$ $subsp.$ $enterica$ among $Houttuynia$ $cordata$, yellow onion and red onion extracts. The in vitro results showed the concentration-dependent effects on the expression of both filaggrin and SPT in HaCat cells among 0.01%, 0.05%, 0.1%, 0.5% extracts in Houttuynia cordata and red onion, reflecting the notion that $Houttuynia$ $cordata$ and red onion can induce epidermal keratinocyte differentiation and improve the recovery of skin barrier functions. The concentration-dependent effects also have been shown on the expression of both COX-2 and AP-1 among 0.01%, 0.05%, 0.1%, 0.5% extracts in $Houttuynia$ $cordata$ and red onion, while slight effect in yellow onion. Conclusion : Red onion juice could be a potential candidate enhanser for the skin care and cosmetology.

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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