• 동의생리병리학회지
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• 제32권2호
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• pp.106-112
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• 2018

The aim of this research was to determine the diverse effects of Sappan Lignum extract and brazilin on human keratinocyte HaCaT cells. We confirmed the antioxidant effect of Sappan Lignum extract and brazilin was analyzed by using an ABTS assay, confirming the efficacy of water extraction method. Also, we examined effect of Sappan Lignum extract and brazilin on the cell viability, using the MTS assay in HaCaT cells. mRNA expression levels of tight junction-related genes associated with skin barrier in HaCaT cells were analyzed using quantitative real-time PCR analysis. Sappan Lignum extract increased the cellular activity of HaCaT cells and the expression of the tight junction-related genes claudin 3, claudin 6, and ZO-2. Brazilin displayed the same effects as that of the extract on HaCaT cells activity and tight junction-related genes expression. Furthermore, dispase assay demonstrated altered cell-cell adhesion strength of Sappan Lignum extract or brazilin treated HaCaT cells. Sappan Lignum extract or brazilin might be an useful ingredient in skin-mosturizinng and anti-wrinkle cosmetics, given its effects of altering mRNA expression of tight junction-related genes and enhancing cell-cell adhesion strength of HaCaT cells.

• Journal of Microbiology and Biotechnology
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• 제31권3호
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• pp.475-482
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• 2021

Prodigiosins, which are natural tripyrrole red pigments and synthetic derivatives, reportedly have multiple biological effects mainly on various types of cancer cells. However, the effects of bacterial prodigiosin on non-cancerous HaCaT human skin keratinocytes have not been reported. Therefore, the present study aimed to investigate the functional activities of prodigiosin derived from cultures of the bacterium Hahella chejuensis in HaCaT cells. Cell viability, the cell proliferation rate, and reactive oxygen species (ROS) production in vitro were assayed following treatment of HaCaT cells with prodigiosin. Prodigiosin did not cause cytotoxicity and notably increased proliferation of HaCaT cells. Furthermore, prodigiosin reduced ultraviolet (UV) irradiation-induced ROS production and the inflammatory response in HaCaT cells. More importantly, prodigiosin reduced matrix metalloproteinase-9 expression and increased collagen synthesis in UV-irradiated HaCaT cells, demonstrating that it elicits anti-aging effects. In conclusion, our results reveal that H. chejuensis-derived prodigiosin is a potential natural product to develop functional cosmetic ingredients.

• BMB Reports
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• 제51권4호
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• pp.188-193
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• 2018

Caffeoylserotonin (CaS), one derivative of serotonin (5-HT), is a secondary metabolite produced in pepper fruits with strong antioxidant activities. In this study, we investigated the effect of CaS on proliferation and migration of human keratinocyte HaCaT cells compared to that of 5-HT. CaS enhanced keratinocyte proliferation even under serum deficient condition. This effect of CaS was mediated by serotonin 2B receptor (5-HT2BR) related to the cell proliferation effect of 5-HT. We also confirmed that both CaS and 5-HT induced G1 progression via 5-HT2BR/ERK pathway in HaCaT cells. However, Akt pathway was additionally involved in upregulated expression levels of cyclin D1 and cyclin E induced by CaS by activating 5-HT2BR. Moreover, CaS and 5-HT induced cell migration in HaCaT cells via 5-HT2BR. However, 5-HT regulated cell migration only through ERK/AP-1/MMP9 pathway while additional Akt/NF-${\kappa}B$/MMP9 pathway was involved in the cell migration effect of CaS. These results suggest that CaS can enhance keratinocyte proliferation and migration. It might have potential as a reagent beneficial for wound closing and cell regeneration.

• 생명과학회지
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• 제29권10호
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• pp.1071-1079
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• 2019

본 연구에서 우리는 오미자 종자 분획물의 항산화 활성과 자외선에 유도된 human HaCaT cell에서의 MMPs의 발현억제에 의한 항노화 효과를 조사하고자 하였다. 전자공여능, ABTS 라디칼 소거분석, 그리고 hydrogen peroxide 소거분석 실험을 통해, 오미자 종자의 분획물 중 에틸아세테이트 분획물(SCEAf)이 가장 우수한 라디칼 소거활성을 가졌고, collagenase 저해 활성 실험에서 SCEAf는 $500{\mu}g/ml$의 농도에서 92.3% 이상의 저해효과를 보였음을 확인하였다. SCEAf의 HaCaT cell에 대한 세포 독성을 확인하기 위해 MTT assay를 실시한 결과, SCEAf는 세포에 독성이 없음을 나타낼 뿐 아니라 UVB에 손상된 세포의 생존율을 증가시킴으로써 세포 활성에 도움을 주는 것을 확인할 수 있었다. SCEAf의 HaCaT cell에서의 항노화 효과를 조사하기 위해 UVB $50mJ/cm^2$에 유도된 HaCaT cell에 SCEAf를 처리한 후 MMP-1과 -3의 발현을 Western blotting과 reverse-transcription polymerase chain reaction으로 분석하였다. 그 결과 SCEAf가 MMP-1과 -3의 단백질과 mRNAs의 발현을 농도의존적으로 억제시켰다. 이러한 결과는 오미자 종자 에틸아세테이트 분획물이 자외선의 손상을 받은 피부 각질형성세포에서의 collageanse 활성을 억제하여 노화를 예방하고 증상을 완화시킬 것으로 기대할 수 있다. 이러한 결과를 바탕으로 우리는 오미자 종자가 화장품과 식품 산업에서 항노화 효과가 있는 기능성 원료로서 사용하기 위한 잠재성을 가질 것으로 기대한다.

• KSBB Journal
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• 제28권6호
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• pp.394-399
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• 2013

Astragalus membranaceus Bunge is used in herbal medicine in Eastern Asian countries including Korea. In this study, we assessed the effects of A. membranaceus extract (AM) on the aquaporin-3 (AQP3) protein expression in HaCaT cells. AM did not affect viability of HaCaT cells. AQP3 expression and cell migration seem to be maximal at $100{\mu}g/mL$ concentration. Epidermal growth factor receptor (EGFR) kinase inhibitor, PD153035, blocked AM-induced AQP3 expression and cell migration. In addition, an 80% ethanol extracts of herbal prescription, SinhyoTakleesan (ST), which is composed of A. membranaceus, Angelicae gigantis, Glycyrrhiza glabra Linne, and Lonicera japonica Flos also induced AQP3 expression at $20{\mu}g/mL$ in HaCaT cells. Collectively, these results suggest that AM induce AQP3 expression via EGFR pathway.

• 대한임상검사과학회지
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• 제47권3호
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• pp.112-116
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• 2015

산화질소(Nitric Oxide, NO)는 생리학적, 병리학적으로 주요한 역할을 하고 있는 것으로 알려져 있다. 본 실험에서는 NO donor인 sodium nitroprusside (SNP)가 HaCaT 세포에서 apoptosis를 유도한다는 것을 DAPI염색과 PARP, caspase-3 단백질 절단을 western blot으로 확인하였다. SNP는 ER stress와 관련 있는 Bip, CHOP 유전자 발현에는 영향이 없었다. 최근 NAD+ 의존 deacetylase인 sirt1이 세포의 생존 및 사멸에 매우 중요한 단백질이라는 보고가 있다. 본 실험에서 SNP는 HaCaT 세포의 sirt1 유전자 발현을 감소시켰으며 이는 apoptosis와 관련이 있을 수 있다.

• 한국응용과학기술학회지
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• 제38권3호
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• pp.870-882
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• 2021

황련은 한의학에서 진정, 소염, 항균 및 해열 등의 여러 질병의 치료에 이용되어져 왔다. 본 연구에서는 UVB와 TNF-α/IFN-γ로 유도된 HaCaT 세포에서 피부장벽과 염증관련 인자에 대한 황련 열수 추출물(CCW)의 효과를 조사하고, 보습 및 항염증 소재로서의 가능성을 알아보고자 하였다. 각 질형성세포인 HaCaT 세포에서 CCW의 보습 및 항염 효과를 확인하기 위해 MTT assay를 통한 시료의 독성 평가를 실시하였다. 이를 바탕으로, HA(Hyaluronic acid) 생성량과 filaggrin의 protein 및 mRNA 발현을 측정하였다. TNF-α/IFN-γ로 자극된 HaCaT 세포에서 CCW는 농도 의존적으로 HA의 생성량을 증가 시켰고, filaggrin의 protein 및 mRNA 발현 측정 결과 CCW의 농도 증가에 따라 발현율도 증가하였다. UVB로 유도된 HaCaT 세포에서 CCW는 ROS의 생성을 감소 시켰고, 양성대조군(positive control)인 EGCG( (-)-epigallocatechin-3-gallate)와 유의한 결과를 보여 주었다. 또한, CCW는 염증성 사이토카인 TNF-α, IL-1β, IL-6, IL-8의 발현을 억제 시켰고, TNF-α/IFN-γ로 자극된 HaCaT 세포에서 피부 염증에 따른 주요 인자인 COX-2의 protein 및 mRNA 발현이 농도 의존적으로 감소되는 것을 확인할 수 있었다. 이러한 결과는, 황련 열수 추출물의 보습 및 항염증 효과를 가지는 화장품 소재로서의 가치를 제안할 수 있다.

• 대한예방한의학회지
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• 제19권3호
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• pp.141-154
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• 2015

Objective : In this paper, we investigated the anti-oxidative capacities and protective effects of water extract of palmul-tang (PMT) against Ultraviolet B(UVB)-induced oxidative damage in human keratinocytes(HaCaT). Method : To evaluate the anti-oxidative activities of PMT, we measured scavenging activities on 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation and reducing power of PMT. To give an oxidative stress to HaCaT cells, UVB was irradiated with $40mJ/cm^2$ to HaCaT cells. To detect the protective effects of PMT against UVB, we measured cell viability, apoptotic bodies and reactive oxygen species(ROS) generation. Results : PMT showed the anti-oxidative activities by scavenging DPPH radical, hydroxyl radical, hydrogen peroxide, superoxide anion, lipid peroxidation. Also PMT showed high reducing values. The UVB-induced oxidative conditions led to the cell apoptosis. However, treatment with PMT reduced oxidative stress conditions, including inhibition of cell apoptosis and expression of ROS. Conclusion : PMT had anti-oxidative activities and exhibited protective effects against UVB on HaCaT cells. PMT would be useful for the development of cosmetics treating UVB-induced skin aging.

• Toxicological Research
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• 제24권1호
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• pp.37-44
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• 2008

The in vitro cell transformation assays (CTA) were performed using BALB/3T3 murine fibroblasts and HaCaT human keratinocytes in order to evaluate concordance between both in vitro CTAs and carcinogenicity with compounds differing in their genotoxic and carcinogenic potential. Six test articles were evaluated, two each from three classes of compounds: genotoxic carcinogens (2-amino-5-nitrophenol and 4-nitroquinoline-N-oxide), genotoxic noncarcinogens (8-hydroxyquinoline and benzyl alcohol), and nongenotoxic carcinogens (methyl carbamate and N-nitrosodiphenylamine). Any foci of size $\geq$2 mm regardless of invasiveness and piling was scored as positive in CTA with BALB/3T3. As expected, four carcinogens regardless of their genotoxicity had positive outcomes in two-stage CTA using BALB/3T3 cells. However, of the two genotoxic noncarcinogens, benzyl alcohol was positive CTA finding. We concluded that, of the 6 chemicals tested, the sensitivity for BALB/3T3 system was reasonably high, being 100%. The respective specificity for BALB/3T3 assay was 50%. We also investigated the correlation between results of BALB/3T3 assay and results from HaCaT assay in order to develop a reliable human cell transformation assay. However, evaluation of staining at later time points beyond the confluency stage did not yield further assessable data because most of HaCaT cells were detached after $2{\sim}3$ days of confluency. Thus, after test article treatment, HaCaT cells were split before massive cell death began. In this modified protocol for this HaCaT system, growing attached colonies were counted instead of transformed foci 3 weeks since last subculture. Compared to BALB/3T3 assay, HaCaT assay showed moderate low sensitivity and high specificity. Despite these differences in specificity and sensitivity, both cell systems did exhibit same good concordance between in vitro CTA and rodent carcinogenicity findings (overall 83% concordant results). At present the major weakness of these in vitro CTA is lack of validation for regulatory acceptance and use. Thus, more controlled studies will be needed in order to be better able to assess and quantitatively estimate in vitro CTA data.

• 치위생과학회지
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• 제7권1호
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• pp.31-35
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• 2007

Caesalpinia sappan L. (Leguminosae) is an oriental medicinal herb distributed in China and Taiwan, and its heartwood has been traditionally used as an analgesic, a therapy for thrombosis or tumor. This study was to investigate the proliferation and inhibition effects of Caesalpinia sappan extracts against human epithelial cell and cancer cell lines. The methanol extract of dried C. sappan heartwood was evaporated (KS-6), and then sequentially extracted by hexane (KS-01), chloroform (KS-02), ethyl acetate (KS-03), n-butanol (KS-04), and water (KS-05). After 48 hr of exposure, these fractions at a concentration of $50{\mu}g/ml$ significantly increased, and reduced cell proliferation in both human normal epithelial and cancer cell lines. The ethyl acetate fraction (KS-03) among organic solvent fractions was 120.2% of the most proliferation activity ($50{\mu}g/ml$) against HaCaT human epithelial cell. However, fractions from chloroform, butanolic and methanolic extract had 7.2, 28.7 and 20.8% of antiproliferative effect on HaCaT cell, respectively. In cell proliferation effects of C. sappan extract on HeLa, SiHa and C33A human cervical cancer cells, chloroform fraction (KS-2) was the most antiproliferative activity, its antiproliferative rate (dosage, $50{\mu}g/ml$) relative to control was 25.8, 12.2 and 17.4% for SiHa, HeLa and C33A, respectively. The results indicated that the six extract fractions could induce cell cycle stimulate or arrest in some way. Finally, further investigation is needed to assess the molecular mechanisms mediated anticancer activities of this plant.