• Bulletin of Food Technology
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• v.12 no.2
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• pp.26-38
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• 1999

조리가공 중 생성되는 주요 발암성 이환방향족아민(heterocyclic aromatic amine)인 Trp-P-1 및 Trp-P-2가 human 및 rat keratinocytes에 대해 나타내는 세포 독성을 colony expansion법의 의해 조사, 비교하였다. 특히 Trp-P-2는 human keratinocytes에 대해서는 독성을 나타내지 않은데 반해 rat keratinocytes(계대수 2-5)에 대해서는 독성을 나타내는 선택성을 나타내었다. 이러한 Trp-P-2의 종(species)간 독성감수성 차이가 대사 효소계 활성이나 mutagenic activation상의 차이에 기인되는지를 살펴본 결과, CYP4501As 및 독성감수성 차이가 크게 나타났던 human 및 rat keratinocytes의 microsome에서 거의 같았다. 이와 같은 결과는 CYP4501A1 및 CYP1B1의 mRNA의 발현정도를 northernblot에 의해 살펴보았던 결과에서도 일치하였다. 반면 Trp-P-2의 대사활성화 및 해독화에 관여하는 효소인 N.O-acetyltransferase(NAT)활성은 rat keratinocytes보다 human keratinocytes에서 높았다. 일반적으로 독성물질의 해독화에 관여하는 glutathione S-transferase(GST) 또한 rat keratinocytes보다 human keratinocytes에서 높게 나타났다. Trp-P-2가 mutagenic metabolite로 활성화되는 정도를 salmonella microsome microsuspension assay로 살펴본 결과, 독성 감수성 차이가 크게 나타났던 human 및 rat keratinocytes간의 활성은 비슷한 것으로 나타났다. DNA 및 단백질 adduct형성능의 경우, human 및 rat keratinocytes간 DNA adduct형성능에는 차이가 없었고, 단백질 adduct형성능의 경우만 Trp-P-2에 대한 독성감수성 정도가 컸던 rat keratinocytes가 다른 세포들에 비해 크게 나타났다. 이상의 결과를 종합해 볼 때, CYP1A- 또는 CYP1B1-관련 마이크로솜 효소활성이나 mutagenic activation은 human 및 rat keratinocytes간에 나타났던 독성 감수성의 차이를 설명할 수 없으며, 해독화에 관여하는 효소활성이 종간 관찰되었던 독성 감수성의 차이에 더 중요한 역할을 하는 것으로 보인다.

• Biomolecules & Therapeutics
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• v.3 no.1
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• pp.80-84
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• 1995

Effects of titrated extract of Centella asiatica (TECA) and epidermal growth factor (EGF) isolated from the urine of pregnant horse on the proliferation of human epidermal keratinocyte in culture were studied. An increase in the number of keratinocyte was observed with the treatment of TECA at the concentration ranges from 1 $\mu\textrm{g}$/mι to 100 $\mu\textrm{g}$/mι. Effects of low molecular weight EGF (LEGF) and high molecular weight EGF (HEGF) on the proliferation of keratinocyte in culture were also studied. The number of keratinocyte in culture was significantly increased with LEGF and HEGF respectively at the concentration of 10 ng/mι. Simultaneous treatment of the keratinocyte with LEGF, HEGF and TECA led to the increased proliferation of keratinocytes resulting 96% of the effect of a positive control, EGF isolated from mouse submaxillary glands.

• Journal of Ginseng Research
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• v.42 no.2
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• pp.229-232
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• 2018

• Toxicological Research
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• v.32 no.4
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• pp.345-351
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• 2016

Propolis is a resinous material collected by honeybees from several plant sources. This research aimed at showing its protective effect against UVA-induced apoptosis of human keratinocyte HaCaT cells. Using Hoechst staining, it was demonstrated that propolis (5 and $10{\mu}g/mL$) significantly inhibited the apoptosis of HaCaT cells induced by UVA-irradiation. Propolis also showed the protective effect against loss of mitochondrial membrane potential induced by UVA-irradiaiton in HaCaT cells. Propolis also inhibited the expression of activated caspase-3 induced by UVA-irradiation. To investigate the role of ROS in UVA-induced apoptosis and protection by propolis, the generation of ROS was determined in cells. The results showed that the generation of ROS was markedly reduced in cells pretreated with propolis. Consequently, propolis protected human keratinocyte HaCaT cells against UVA-induced apoptosis, which might be related to the reduction of ROS generation by UVA-irradiation.

• Nutrition Research and Practice
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• v.11 no.4
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• pp.275-280
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• 2017

BACKGROUND/OBJECTIVES: Re-epithelialization has an important role in skin wound healing. Astaxanthin (ASX), a carotenoid found in crustaceans including shrimp, crab, and salmon, has been widely used for skin protection. Therefore, we investigated the effects of ASX on proliferation and migration of human skin keratinocyte cells and explored the mechanism associated with that migration. MATERIAL/METHOD: HaCaT keratinocyte cells were exposed to $0.25-1{\mu}g/mL$ of ASX. Proliferation of keratinocytes was analyzed by using MTT assays and flow cytometry. Keratinocyte migration was determined by using a scratch wound-healing assay. A mechanism for regulation of migration was explored via immunocytochemistry and western blot analysis. RESULTS: Our results suggest that ASX produces no significant toxicity in human keratinocyte cells. Cell-cycle analysis on ASX-treated keratinocytes demonstrated a significant increase in keratinocyte cell proliferation at the S phase. In addition, ASX increased keratinocyte motility across the wound space in a time-dependent manner. The mechanism by which ASX increased keratinocyte migration was associated with induction of filopodia and formation of lamellipodia, as well as with increased Cdc42 and Rac1 activation and decreased RhoA activation. CONCLUSIONS: ASX stimulates the migration of keratinocytes through Cdc42, Rac1 activation and RhoA inhibition. ASX has a positive role in the re-epithelialization of wounds. Our results may encourage further in vivo and clinical study into the development of ASX as a potential agent for wound repair.

• Korean Journal of Pharmacognosy
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• v.46 no.2
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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.

• Biomedical Science Letters
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• v.8 no.4
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• pp.229-234
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• 2002

Chlorella is rich in chlorella growth factor (CGF). A review of the literature has described that CGF improves the capability of a Th1-based immunity, anticancer, antioxidant antibacterial activity, growth promotion, wound healing and so on, but has not studied the effect for the metabolism and the proliferation of human skin keratinocyte. The aim of this study was to examine the effect of metabolism and the proliferation of human skin keratinocyte in vitro. CGF was extracted with an autoclaving method which is a modified hot-water extraction method from dried chlorella and conformed by means of absorbance 0.22 at 260 nm. We have measured the extracellular acidification rate (ECAR) of the CGF by Cytosensor$^{\circledR}$ Microphysiometer and evaluated responsiveness depending upon the dosage on the HaCaT cell. The ECAR for the concentrations of 0.15, 1.5, 15, 150 $\mu\textrm{g}$/ml of CGF increased as a 103.6, 128.2, 149.0 and 423.9%, respectively compared to control (0.0 $\mu\textrm{g}$/ml, 100% ECAR). The ECAR for ErbBl tyrosine kinase inhibited by 4-anilinoquinazolines, $C_{16}$H$_{14}$BrN$_3$O$_2$.HCl on tile HaCaT cells with the amounts of 10 $\mu\textrm{g}$/ml of the CCF compared with 100 $\mu\textrm{g}$/ml of rhEGF. The conclusion of the study is that CGF might increase human epidermal keratinocyte proliferation through the interaction between the epidermal growth factor receptor and itself.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.5
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• pp.814-821
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• 1997

Kimchi is composed of many ingredients such as Chinese cabbage, garlic, ginger, and red pepper and fermented fish extract. Some of them were known to have antioxidative activities due to their scavenging effect against reactive oxygen species(ROS). To study the health effects of kimchi on human skin cells, keratinocyte(A431, epidermoid carcinoma, human) and fibroblast(CCD-986SK, normal control, human) were cultured in oxidative stress condition provoked by paraquat, a superoxide anion generator, and hydrogen peroxide in the absence and presence of kimchi extract. The survival rate of keratinocyte was greatly reduced when exposed over 1mM concentration of hydrogen peroxide($H_{2}O_{2}$), but cytotoxicity of $H_{2}O_{2}$ was significantly reduced by kimchi extracts on cells. Especially 2 week-fermented kimchi decreased remarkably the cytotoxicity by $H_{2}O_{2}$ to keratinocyte cells. Over 1mM of paraquat concentration showed strong cell toxicity on keratinocyte, but the extracts from kimchi fermented for 1, 2 and 3 weeks showed protective effects in order. Fibroblast cells were significantly affected by $H_{2}O_{2}$ as were keratinocyte cells. Although almost all extacts of kimchi of different fermentation periods showed protective effect against cell killing at 0.5mM concentration of $H_{2}O_{2}$ week-fermented kimchi extract showed the strongest protective effect on fibroblast cells treated with 1mM $H_{2}O_{2}$ for either 1 day or 4 days. However most of kimchi extracts showed weak preventive effect or no effect on oxidative stress produced by paraquat. In conclusion, 2 week-fermented kimchi extract seems to have the best potential in preventing skin cells against oxidative damage which might be related to their scavenging effects of kimchi components produced during their fermentation process.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.4
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• pp.329-335
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• 2017

We have studied on the keratinocytes differentiation and skin barrier function using Adenophorae radix (A. radix) root extract, which was known to contain triterpenoid, saponin and starch. A. radix root extracts showed the $PPAR{\alpha}$ expression level of Wy-14,643 $0.5-1.0{\mu}M$ in CV-1 cells. The cornified envelop formation (CE) of human keratinocyte cell line (HaCaT) and normal human keratinocyte (NHK) showed a statistically significant increased compared to the control. When HaCaT cells were treated with A. radix root extract, transglutaminase (TGase-1) was significantly increased. As a result of clinical study of the simple cosmetic formulation containing A. radix root extract for about 2 weeks, TEWL values were significantly decreased and water contents were increased. The ceramides, which were obtained from the inner forearm, were also significantly increased statistically. We suggest that the A. radix root extract can be used as a preventive and therapeutic agent for skin diseases such as dry skin and atopy.

• Archives of Plastic Surgery
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• v.35 no.2
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• pp.127-133
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• 2008

Purpose: The purpose of this study is to evaluate the remodeling process of the various skin substitutes in 4th and 6th weeks following the transplantation when transplanted onto nude mice. Methods: Three types of artificial skin substitutes, such as PLGA scaffold with keratinocyte sheets(group 1), acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), bioengineered skin($Neoderm^{(TM)}$)(group 3), were applied to the wound on nude mice. All mice were killed in 2, 4 weeks and/or 6 weeks after grafting and tissue samples were harvested from the back of mice. The changes in wound size, degree of angiogenesis, formation of basement membrane and epidermis, density of collagen fibers and neural restoration were examined. Results: There was no significant changes in wound size among the three groups. However, the size of wound decreased in the non-substituted group due to contracture. Degree of angiogenesis and systhesis of collagen or neurofilaments were mostly increased in bioengineered skin($Neoderm^{(TM)}$)(group 3), followed by acellular human dermis($Surederm^{(TM)}$) and keratinocyte sheet(group 2), PLGA scaffold with keratinocyte sheets (group 1). However, group 3 and group 2 showed similar thickness of basement membrane and epidermis. Conclusion: We found that degree of angiogenesis, formation of basement membrane and skin appendages, density of collagen fibers and neurofilaments can be the categories to evaluate the success of artificial skin substitution in early stages.