• 생약학회지
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• 제46권2호
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• pp.116-122
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• 2015

Keratinocytes are first barrier against outer challenges on skin. However, it is still largely unknown about effective protectors against ultraviolet B (UVB), and oxidative stress in human keratinocyte, HaCaT cells. Inducible heme oxygenase (HO)-1 acts against oxidants that are thought to play a role in the pathogenesis of skin disorders. Therefore, the purpose of this study was to evaluate the effect of Portulaca oleracea 70% EtOH extracts against hydrogen peroxide (H₂O₂)-induced oxidative stress in human keratinocytes, HaCaT cells. P. oleracea 70% EtOH extracts showed the potent protective effects on H₂O₂-induced toxicity by induced the expression of HO-1 in human keratinocyte, HaCaT cells. Furthermore, P. oleracea 70 % EtOH extracts caused the nuclear accumulation of nuclear factor E2-related factor 2 (Nrf2) in human keratinocytes, HaCaT cells. In addition, we found that treatment with c-Jun N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea 70% EtOH extracts-induced HO-1 expression, and JNK inhibitor (SP600125) also inhibited protective effects by P. oleracea 70% EtOH extracts. Therefore, these results suggest that P. oleracea 70 % EtOH extracts increases cellular resistance to H₂O₂-induced oxidative injury in human keratinocyte, HaCaT cells, presumably through JNK pathway-Nrf2-dependent HO-1 expression.

• 동의생리병리학회지
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• 제21권4호
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• pp.1017-1024
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• 2007

Thymus and activation-regulated chemokine (TARC/CCL-17), produced by keratinocytes, is a CC chemokine known to selectively Th2 type T cells via $CCR4^+$ and is implicated in the development of atopic dermatitis (AD). TARC/CCL17 expression was induced by cytokines such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$). We recently found that the wogonin, a flavone isolated from Scutellaria baicalensis, suppressed TARC expression via heme oxygenase 1 (HO1) in human keratinocytes induced with mite antigen. However, little is known about the inhibitory mechanism of wogonin on TARC/CCL-17 expression stimulated with cytokines. To investigate the inhibitory mechanism, I determined the inhibitory effects of wogonin on the activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and $I{\kappa}B{\alpha}$ phosphorylation, and also examined the activation of p38 MAP kainase in HaCaT keratinocytes stimulated with TNF-${\alpha}$ and IFN-${\gamma}$. Wogonin inhibited NF-${\kappa}B$-DNA complex, NF-${\kappa}B$ binding activity, and the phosphorylation of $I{\kappa}B{\alpha}$ in a dose dependent manner. Wogonin also inhibited the translocation of NF-${\kappa}B$ from cytosol to nucleus. Moreover, the phosphorylation of of p38 MAP kinase in the TNF-${\alpha}$ and IFN-${\gamma}$-stimulated HaCaT keratinocytes were suppressed by wogonin in a dose dependent manner. These results suggest that wogonin may inhibit cytokine-induced NF-${\kappa}B$ activation by $I{\kappa}B{\alpha}$ degradation via suppression of p38 MAP kinase signaling pathway in keratinocytes and modulation of wogonin signaling pathway may be beneficial for the treatment of AD.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.273.2-274
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• 2002

Human epidermal keratinocytes consist of stem cells. transit amplifying cells. and postmitotic differentiating cells. Among them, stem cells playa critical role in cell renewal. wound healing. and neoplasia. However. till now, specific markers of human epidermal keratinocytes are not clearly defined. In the present study. we separated putative stem cells from other cells using fluorescence activated cell sorting (FACS). based on differences in a6-integrin and CD71 expression. (omitted)

• Journal of Ginseng Research
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• 제43권2호
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• pp.300-304
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• 2019

• 한국동물학회:학술대회논문집
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• 한국동물학회 1997년도 한국생물과학협회 학술발표대회
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• pp.208.1-208
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• 1997

No Abstract, See Full Text

• 대한본초학회지
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• 제29권6호
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• pp.63-71
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• 2014

Objectives : Lonicerae japonicae Flos(LJF) has been reported to exhibit anti-oxidant, anti-inflammatory, anti-viral, anti-rheumatoid properties. However, it is still largely unknown whether LJF inhibits the ultraviolet(UV)B-induced oxidative damage in human HaCaT keratinocytes. Therefore in this paper, we investigated the anti-oxidative capacity and protective effect of LJF against UVB-induced oxidative demage in human HaCaT keratinocytes. Methods : To evaluate the anti-oxidative activity of LJF extracts, we measured total phenolic contents, total flavonoid contents, antioxidant capacity, and superoxide scavenging activity. To give an oxidative stress to HaCaT cells, UVB was irradiated with $200mJ/cm^2$ to HaCaT cells. To detect the protective effect of LJF against UVB, we measured cell viability, DNA fragmentation and reactive oxygen species (ROS) production. In addition, we performed high-performance liquid chromatography (HPLC) analysis to find a major component of LJF. Results : LJF contained phenolic and flavonoid contents, and showed the anti-oxidant and superoxide scavenging activity. The UVB-induced oxidative conditions led to the cell death, DNA fragmentation and reactive oxygen species (ROS) production. However, pretreatment with LJF reduced oxidative conditions, including inhibition of cell death, DNA fragmentation and ROS production. In addition, we found out chlorogenic acid as major component of LJF. Conclusions : These results could suggest that LJF contained anti-oxidative contents and exhibited protective effects against UVB on human HaCaT keratinocytes. And the effective compound of LJF which could show protective activities against UVB is chlorogenic acid. Thus, LJF and chlorogenic acid would be useful for the development of drug or cosmetics treating skin troubles.

• 생명과학회지
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• 제28권8호
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• pp.892-899
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• 2018

쇠비름(Portulaca oleracea.L)은 쇠비름과에 속하는 한해살이풀로서 리놀렌산과 같은 불포화지방산, 페놀성 화합물, 플라보노이드, 비타민 C, 미네랄 함량이 높은 것으로 보고되어 있다. 본 연구에서는 쇠비름 추출물을 이용하여 UVB를 조사한 인간각질형성세포에서 광노화 억제능을 확인하였다. Matrix metalloproteinases는 세포의 기질을 분해하는 효소로 MMP-1는 collagenase, MMP-2와 MMP-9는 gelatinases로 피부 진피층을 구성하는 type I collagen을 분해시키는데 영향을 미친다. UVB를 조사한 인간각질형성세포에서 쇠비름 추출물을 처리했을 때 MMP-1, -2, -9의 발현이 감소하였으며, type I procollagen의 발현은 증가하는 것으로 나타났다. 또한 쇠비름 추출물을 처리한 군에서 UV에 의한 ROS 생성이 감소하였는데 이는 Nrf-2의 활성화를 통한 항산화 인자 SOD-1과 OH-1의 발현 증가로 인해 세포내 ROS 생성이 감소한 것으로 사료된다. 따라서 본 연구 결과를 통해 쇠비름 추출물이 UVB를 조사한 인간각질형성세포에서 MMP 인자 및 항산화 인자의 발현 조절을 통해 광노화로부터의 세포 보호 능을 가지는 것을 확인하였으며 나아가 화장품 소재로서의 개발 가능성을 확인하였다.

• 대한본초학회지
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• 제26권2호
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• pp.45-49
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• 2011

Objectives : The purpose of this study was to investigate the anti-aging effects on cultured human keratinocytes with Nelumbo nucifera extracts. Methods : Each parts of leaves, flowers and stamen were extracted with water or 70% ethanol. These extracts were tested for cell viability on HaCaT cells (human keratinocyte line) by MTT assay. We investigated the effects of Ultraviolet-B (UVB) irradiation on cytotoxicity and lipid peroxidation in cultured skin keratinocytes. Results : The ethanol extract of Nelumbo nucifera flowers showed maximun cell viability as 111.39% in 30 ug/ml concentration. The water extracts of stamen, flowers, leaves showed cell viability as 107.12, 101.65, 101.46%, respectively. HaCaT keratinocytes were survived 63.06% at $20mJ/cm^2$ UVB irradiation. The cell membrane lipid peroxidation was measured by accumulation malondialdehyde (MDA). The levels of MDA were decreased by the ethanol extract of Nelumbo nucifera flowers and the water extracts of stamen. Conclusions : These finding suggest that the ethanol extract of Nelumbo nucifera flowers prevent anti-aging effects on cultured human keratinocytes during UVB irradiation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권3호
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• pp.211-220
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• 2007

Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

• KSBB Journal
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• 제29권1호
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• pp.42-49
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• 2014

The aim of this study was to develop a composite human skin equivalent for wound healing. Collagen type1 and acellular dermal matrix powder were utilized as the scaffold with dermal fibroblasts and keratinocytes for the development of a composite human skin equivalent. Fibroblast maintained the volume of composite skin equivalent and also induced keratinocytes to attach and proliferate on the surface of composite skin equivalent. The composite human skin equivalent had a structure and curvature similar to those of real skin. Balb-C nu/nu mice were used for the evaluation of full-thickness wound healing effect of the composite human skin equivalent. Graft of composite skin equivalent on full-thickness wound promoted re-epithelialization and granulation tissue formation at 9 days. Given the average wound-healing time (14 days), the wound in the developed composite skin equivalent healed quickly. The overall results indicated that this three-dimensional composite human skin equivalent can be used to effectively enhance wound healing.