• 한국응용과학기술학회지
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• 제41권2호
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• pp.459-471
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• 2024

현대사회에서 스트레스와 긴장감은 피할 수 없는 요인이다. 다양한 피부질환은 스트레스를 일으키는 중요한 요인으로 언급되고 있다. 피부질환을 가진 환자들은 수면상태가 원활하지 않아 전반적으로 수면 효율이 낮다. 또한 피부질환으로 인해 심리적 스트레스 수치가 높아지고, 이와 같은 과정은 반복적으로 발생하고 있다. 피부질환과 스트레스는 상호적으로 연관되어 있으며, psychodematology에 대한 연구가 증가하고 있다. 이에 본 연구에서는 피부질환을 저하 시킬 수 있는 호박, 작약, 타트체리 복합물을 활용하여 피부 각질 형성 세포에서 스트레스로 인한 만성 피부질환을 개선할 수 있는 소재를 개발하고 효능을 입증하고자 하였다. HaCaT 각질형성세포에 복합 추출물은 12.5, 25, 50, 100 ㎍/mL 농도 의존적으로 TNF-α, IL-1β, IL-6, MDC, TARC 발현량이 저해되었으며 특히 IL-1β의 경우, 100 ㎍/mL의 농도에서 40% 이상 저해하는 우수한 효능을 확인하였다. 또한 AQP-3, HA, filaggrin의 생성량 농도 의존적으로 유의미한 증가를 보이며 TNF-α/IFN-γ로 증가된 p-ERK, p-JNK, p-p38의 단백질 발현은 복합 추출물의 처리로 유의하게 감소시 키는 것으로 나타났다. 이를 통하여 해당 복합 추출물은 피부질환을 치료 및 예방할 수 있는 소재로서 활용가치가 있는 것으로 판단되며, 이는 피부질환과 스트레스 간의 상호 관계의 악영향을 낮춰 줄 것으로 판단된다.

• 동의생리병리학회지
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• 제28권5호
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• pp.520-529
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• 2014

The purpose of this study is to investigate the protective effects of Yukmijiwhang-tang(YM) water extracts against the UVB irradiation on the human keratinocyte HaCaT cells. We observed the effects of YM on the oxidative stress, gene expression of pro-inflammatory cytokine such as TNF-${\alpha}$ and IL-$1{\beta}$, and matrix metalloproteinase-9 in UVB-irradiated HaCaT cells. On the effects of oxidative stress and antioxidant function on the treatment with YM, The activity of xanthine oxidase(XO) was significantly decreased by treatment of YM in all the concentrations(p<0.01). The activity of superoxide dismutase(SOD) and catalase(CAT) was significantly increased by treatment of YM in a dose dependent manner(p<0.05 and p<0.01). DPPH radical was erased by treatment of YM under dose of $500{\mu}g/m{\ell}$ concentration. Treatment of HaCaT cells with YM had also significantly reduced intracellular ROS produced by UVB irradiation in a dose dependent manner(p<0.05, p<0.01, p<0.001). Gelatin zymography assay showed that YM downregulated the MMP-9 activity in UVB-irradiated HaCaT cells. RT-PCR analysis revealed that YM suppressed the expression of IL-$1{\beta}$ and MMP-9 however, it has no effects on the expression of TNF-${\alpha}$ and MMP-3. Our study suggests that Yukmijiwhang-tang exert protective actions on the UVB-irradiated HaCaT cells largely by anti-oxidative and anti-inflammatory processes.

• Nutrition Research and Practice
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• 제10권4호
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• pp.371-376
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• 2016

BACKGROUND/OBJECTIVES: Chronic ultraviolet (UV) exposure-induced reactive oxygen species (ROS) are commonly involved in the pathogenesis of skin damage by activating the metalloproteinases (MMP) that break down type I collagen. Adenophora remotiflora (AR) is a perennial wild plant that inhabits Korea, China, and Japan. The present study investigated the protective effects of AR against UVB-induced photo-damage in keratinocytes. MATERIALS/METHODS: An in vitro cell-free system was used to examine the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and nitric oxide (NO). The effect of AR on ROS formation, antioxidant enzymes, elastase, MMP-1 level, and mRNA expression of MMP-1 were determined in UVB-irradiated human keratinocyte HaCaT cells. RESULTS: AR demonstrated strong DPPH free radical and NO scavenging activity in a cell-free system exhibiting $IC_{50}$ values of 1.88 mg/mL and 6.77 mg/mL, respectively. AR pretreatment dose-dependently attenuated the production of UVB-induced intracellular ROS, and antioxidant enzymes (catalase and superoxide dismutase) were enhanced in HaCaT cells. Furthermore, pretreatment of AR prevented UVB-induced elastase and collagen degradation by inhibiting the MMP-1 protein level and mRNA expression. Accordingly, AR treatment elevated collagen content in UVB-irradiated HaCaT cells. CONCLUSION: The present study provides the first evidence of AR inhibiting UVB-induced ROS production and induction of MMP-1 as a result of augmentation of antioxidative activity in HaCaT human keratinocytes. These results suggest that AR might act as an effective inhibitor of UVB-modulated signaling pathways and might serve as a photo-protective agent.

• 대한예방한의학회지
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• 제22권2호
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• pp.109-123
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• 2018

Purpose : In this study, we intended to observe the anti-wrinkle and moisturizing effects of dried pomegranate juice concentration powder (PCP) using in vitro test. Materials and methods : Antioxidant effects of PCP were determined by free radical scavenging capacity (DPPH assay) and the cytotoxicity of PCP was examined in human keratinocyte (HaCaT) and human primary dermal fibroblast-neonatal (HDF) cells. To investigate the moisturizing effect of PCP, hyaluronan synthesis was examined in HaCaT cells. Activity of procollagen production were assessed in HDF cells and elastase inhibition properties of PCP were evaluated in cell free condition, to determine their anti-wrinkle effects. Metalloproteinase 1 (MMP-1) activity was also assessed following UVB irradiation, in the current in vitro experiment. Results : No PCP treatment related significant cytotoxic effects were demonstrated against to the both HDF and HaCaT cells. PCP showed favorable free radical scavenging activities in dose-dependent manner. In PCP-treated HaCaT cells, hyaluronan synthesis was non-significantly but markedly increased, and pro-collagen productions were significantly increased in HDF cells, at all three different concentrations (0.25, 0.75 and 1 mg/ml), and elastase inhibitory activities were observed by PCP treatment. A significant decrease in UVB-induced MMP-1 activity was also observed in 1 mg/ml PCP-treated HDF cells as compared to those of UVB-exposed cells. Conclusions : Taken together, these results suggest that PCP has favorable antioxidant, anti-wrinkle and moisturizing effects without meaningful cytotoxicity on HDF and HaCaT cell lines.

• 대한본초학회지
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• 제36권5호
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• pp.93-99
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• 2021

Objective : Caryophylli Flos has been used in Korean medicine to relieve vomiting and pains caused by chills that make fluid circulation difficult. This study was designed to investigate the protective effect of ethanol extract of Caryophylli Flos (CF) in hydrogen peroxide (H₂O₂)-induced apoptotic cell death in human keratinocyte HaCaT cells. Methods : CF was prepared by extracting 200 g of Caryophylli Flos in 2 L of ethanol for 48 h. Cell viability was measured by MTT assay, and the protein expression was monitored by Western blot analysis. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Reactive oxygen species (ROS) was measured using fluorescent dye, and reduced glutathione (GSH) was determined with a colorimetric commercial kit. Results : CF protected HaCaT cells from cell death caused by oxidative stress after H₂O₂ treatment. H₂O₂ amplified generation of ROS and induced depletion of GSH, whereas these changes in ROS and GSH were inhibited by GF treatment. In addition, H₂O₂ resulted in apoptosis as assessed by TUNEL assay and the expression of apoptosis regulator proteins. However, cells treated with CF showed a decrease in TUNEL-positive cells and restored the reduced expression of procaspase-9, -3 and PARP. Conclusion : This study showed cytoprotective effects of CF by anti-apoptotic activity while exerting antioxidative activity in H₂O₂-treated HaCaT cells. These results suggest that CF could be beneficial in skin damage caused by oxidative stress.

• Journal of Ginseng Research
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• 제46권4호
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• pp.536-542
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• 2022

Background: In aged skin, reactive oxygen species (ROS) induces degradation of the extracellular matrix (ECM), leading to visible aging signs. Collagens in the ECM are cleaved by matrix metalloproteinases (MMPs). Syringaresinol (SYR), isolated from Panax ginseng berry, has various physiological activities, including anti-inflammatory action. However, the anti-aging effects of SYR via antioxidant and autophagy regulation have not been elucidated. Methods: The preventive effect of SYR on skin aging was investigated in human HaCaT keratinocytes in the presence of H₂O₂, and the keratinocyte cells were treated with SYR (0-200 ㎍/mL). mRNA and protein levels of MMP-2 and -9 were determined by real-time PCR and Western blotting, respectively. Radical scavenging activity was researched by 2,2 diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. LC3B level was assessed by Western blotting and confocal microscopy. Results: SYR significantly reduced gene expression and protein levels of MMP-9 and -2 in both H₂O₂-treated and untreated HaCaT cells. SYR did not show cytotoxicity to HaCaT cells. SYR exhibited DPPH and ABTS radical scavenging activities with an EC₅₀ value of 10.77 and 10.35 ㎍/mL, respectively. SYR elevated total levels of endogenous and exogenous LC3B in H₂O₂-stimulated HaCaT cells. 3-Methyladenine (3-MA), an autophagy inhibitor, counteracted the inhibitory effect of SYR on MMP-2 expression. Conclusion: SYR showed antioxidant activity and up-regulated autophagy activity in H₂O₂-stimulated HaCaT cells, lowering the expression of MMP-2 and MMP-9 associated with skin aging. Our results suggest that SYR has potential value as a cosmetic additive for prevention of skin aging.

• 대한화장품학회지
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• 제44권3호
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• pp.343-347
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• 2018

화장품에 방부제(살균/보존제)로 사용되는 1, 2-hexanediol (HD)로 인한 부작용을 극복하기 위하여, Escherichia coli (E. coli)의 ${\beta}-galactosidase$ (${\beta}-gal$)를 이용하여 transgalactosylation 반응으로 1, 2-hexanediol galactoside (HD-Gal)를 합성하였다. 본 연구에서는 합성된 HD-Gal의 인간 피부세포에 대한 독성이 어느 정도인지를 HD와 비교하여 관찰하였다. HD-Gal과 HD의 세포독성은 인간 피부각질형성세포(HaCaT cell line)에 HD와 HD-Gal을 처리한 후, cell proliferation assay를 이용하여 비교 분석하였다. 또한 이때, 위상차 현미경으로 HD-Gal과 HD로 처리한 세포의 상태를 비교 관찰하였다. 그 결과, HD-Gal은 42.2 mM에서 211 mM의 농도 범위에서 세포독성이 관찰되지 않았으며, 현미경 관찰에서도 큰 변화를 관찰할 수 없었다. 그러나, HD의 경우에는 저농도에서(42.2 mM and 84.4 mM)는 세포독성이 관찰되지 않았으나, 고농도(168.8 and 211 mM)에서 매우 높은 세포독성을 나타내었고, 현미경 관찰에서는 고농도에서는 물론이고, 세포독성이 관찰되지 않은 HD의 저농도에서도 세포모양과 세포 수에서의 변화가 관찰되었다. 앞으로 세포독성이 감소된 HD-Gal이 HD의 대체제로서 안전, 건강 및 웰빙 개념의 새로운 용도로 개발될 수 있을 것으로 생각된다.

• 생명과학회지
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• 제29권10호
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• pp.1164-1170
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• 2019

본 연구는 은행외종피 추출물의 항산화 및 DNA 손상 보호용 조성물에 관한 것으로 HaCaT 세포에서 은행외종피 물과 메탄올 추출물의 과산화수소의 공격에 대한 항산화 효과 및 DNA 손상보호에 대한 것이다. 이를 위해 은행외종피 물 추출물(GOSWE)와 은행외종피 메탄올 추출물(GOSME)의 항산화력을 위해서는 DPPH와 과산화수소 소거능을 실시하였다. GSOWE와 GOSME는 DPPH 소거능에서 다소 차이를 보였다. 더욱이, GOSME는 과산화수소 소거능에서 GOSWE보다 탁월한 효과를 발휘하였다. 1 mM $H_2O_2$ 처리된 HaCaT 세포의 세포생존실험에서 GOSME는 세포생존력을 향상시켰다. GOSME는 1 mM $H_2O_2$에 의한 플라스미드 DNA 단편화 저해능과 HaCaT세포에서 게놈 DNA 단편화 억제능이 있는 것으로 확인되었다. 이러한 결과는 GOSME는 활성산소제거를 통한 산화적 스트레스관련 세포손상 억제를 통한 세포사멸억제 및 과산화수소 유발 DNA 손상을 억제 할 수 있다는 것을 제시하였다. 결론적으로 GOSME는 산화적스트레스에 의한 피부질환에 대한 치료 및 방어제로 이용 될 수 있는 신소재로 평가 할 수 있다.

• 대한본초학회지
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• 제34권1호
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• Molecular & Cellular Toxicology
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• 제5권1호
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• pp.51-57
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• 2009

Quantum Dot (QD) nanoparticles are used in various industrial applications, such as diagnostic, drug delivery, and imaging agents of biomedicine. Although QDs are extensively used in many medical science, several studies have been demonstrated the potential toxicity of nanoparticles. The first objective of this study was to investigate the nanotoxicity of QDs in the HaCaT human keratinocyte cell line by focusing on gene expression pattern. In order to evaluate the effect of QDs on gene expression profile in HaCaT cells, we analyzed the differential genes which related to oxidative stress and antioxidant defense mechanisms by using human cDNA microarray and PCR array. A human cDNA microarray was clone set, which was sorted for a list of genes correlated with cell mechanisms. We tried to confirm results of cDNA microarray by using PCR array, which is pathway-focused gene expression profiling technology using Real-Time PCR. Although we could not find the exactly same genes in both methods, we have screened the effects of QDs on global gene expression profiles in human skin cells. In addition, our results show that QD treatment somehow regulates cellular pathways of oxidative stress and antioxidant defense mechanisms. Therefore, we suggest that this study can enlarge our knowledge of the transcriptional profile and identify new candidate biomarker genes to evaluate the toxicity of nanotoxicology.