• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.128-128
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• 2018

Nypa fruticans wurmb (NF) has been used as traditional medicinal food in Asian countries. Especially, NF has been used for conventional medicine to treat inflammatory periodontal diseases. Previous studies have been shown that NF has large amount of useful constituents such as phenolic acids, polyphenols and flavonoids. Also, NF is known as having medicinal effects such as anti-oxidant, anti-inflammatory and cholesterol-lowering effects. NF has recently been attracted to use complementary medicinal food on inflammatory diseases in Korea. However, there are no obvious effects in inflammatory and metabolic diseases also mechanisms has been studied yet. The purpose of this study was to investigate the anti-inflammatory effects of NF and steamed-NF (SNF), which recently has been used as health food, using Human keratinocyte cell line (HaCaT) and Human mast cell line (HMC-1). The cytotoxicities of NF and SNF were measured by using MTT assays in HaCaT cells and HMC-1 cell. To evaluate anti-inflammatory effects of NF and SNF, HaCaT cells were stimulated with tumor necrosis factor $(TNF)-{\alpha}$ and Interferon $(IFN)-{\gamma}$. Also, HMC-1 cells were stimulated with phorbol-12-myristate-13-acetate (PMA) and A23187 calcium ionophore (A23187) to induce allergic inflammation. Inflammatory cytokine were measured by enzyme-linked immunosorbent assay (ELISA). In this result, the extract of NF and SNF (0.01 - 1mg/ml) did not show cytotoxicity in HaCaT cells and HMC-1 cells. In addition, the NF and SNF suppressed the production of interleukin (IL)-6 and IL-8 in HaCaT cells at highest concentration. Furthermore, the treatment of SNF significantly inhibited the secretion level of IL-8 in PMA plus A23187-stimulated HMC-1 cells compared with NF treatment group. These results suggest that the extract of NF and SNF may serve as a potential therapy for skin inflammatory diseases.

• Biomolecules & Therapeutics
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• 제24권6호
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• pp.616-622
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• 2016

Baicalein (5,6,7-trihydroxy-2-phenyl-chromen-4-one) is a flavone, a type of flavonoid, originally isolated from the roots of Scutellaria baicalensis. This study evaluated the protective effects of baicalein against oxidative damage-mediated apoptosis induced by ultraviolet B (UVB) radiation in a human keratinocyte cell line (HaCaT). Baicalein absorbed light within the wavelength range of UVB. In addition, baicalein decreased the level of intracellular reactive oxygen species (ROS) in response to UVB radiation. Baicalein protected cells against UVB radiation-induced DNA breaks, 8-isoprostane generation and protein modification in HaCaT cells. Furthermore, baicalein suppressed the apoptotic cell death by UVB radiation. These findings suggest that baicalein protected HaCaT cells against UVB radiation-induced cell damage and apoptosis by absorbing UVB radiation and scavenging ROS.

• 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.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.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.307-311
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• 2001

The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

• 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.

• 대한본초학회지
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• 제37권3호
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• pp.41-47
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• 2022

Objectives : Terminalia chebula (TC) has been used as a traditional remedy to treat gastrointestinal infectious and inflammatory diseases. However, its protective effects and mechanisms against skin inflammation have not been well-elucidated. Thus, the aim of this study is to evaluate the protective effects of the TC water extract and also to suggest a putative mechanism of TC against skin injury on human keratinocytes, HaCaT cells. Methods : HaCaT cells were pre-treated with TC for 1 h and then stimulated with tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) (10 ng/mL each) to induce skin inflammation and injury. After 24 h, the cells were harvested to evaluate the expression of Th2 chemokines, such as C-C motif chemokine ligand 5 (CCL5, also known as RANTES), C-C chemokine ligand 17 (CCL17, also known as TARC) and C-C chemokine ligand 22 (CCL22, also known as MDC). To investigate the regulatory mechanisms of TC, we also assessed the phosphorylation of signal transducer and activator of transcription 1 (STAT1) signaling pathways in HaCaT cells. Results : Treatment of TC decreased the mRNA levels of RANTES, TARC and MDC with a concentration dependent manner against co-stimulation of TNF-α and IFN-γ. In addition, TC significantly reduced TNF-α and IFN-γ induced phosphorylation of STAT1. Conclusions : In summary, we propose that TC may be a promising candidate for anti-inflammatory skin protector through the inhibition of chemokines via STAT1 deactivation.

• 생명과학회지
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• 제29권5호
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• pp.514-523
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• 2019

인삼(Panax ginseng C. A. Meyer)의 사포닌 진세노사이드 Rb1이 처리된 인간 피부각질세포 HaCaT에서 microarray 분석 및 발현이 증가된 세포사멸 반응에 대한 작용기전을 연구하였다. HaCaT 세포에 진세노사이드 Rb1의 처리로 세포사멸, 유사분열 세포주기의 G2/M 전이, 세포분열, 핵분열, 그리고 단백질 수송 등의 작용기전에 관여하는 유전자들이 2 배 이상 발현이 증가된 것으로 나타났으며, DNA 수선, 감수 핵분열, 그리고 세포외기질 체계 등의 작용기전에 관여하는 유전자들은 2 배 이상 발현이 감소된 것으로 나타났다. 특히 세포사멸 신호전달은 FAS와 PLA2G4A를 경유하는 것으로 나타났으며, 이들 유전자의 상위 조절자로 STAT3가 예측되었다. 세포사멸 반응 경유 유전자 FAS와 PLA2G4A의 활성을 qPCR로 확인한 결과, FAS 유전자는 $10{\mu}g/ml$의 진세노사이드 Rb1를 24시간 동안 처리하였을 경우 약 2 배의 발현 증가와, PLA2G4A 유전자는 6시간 처리부터 약 2 배로 증가되어 24시간 동안 처리시 2 배 이상의 유전자 발현이 증가되었다. 한편 STAT3-siRNA를 이용한 knock-down 실험에서 FAS의 발현 감소와 PLA2G4A의 발현 증가로 상위 조절자 STAT3로부터 FAS 만을 경유하는 것을 알 수 있었다. 이상의 결과 진세노사이드 Rb1의 처리에 의해 상위 조절자인 STAT3는 FAS를 경유하여 세포사멸을 유도하는 것임을 알 수 있다.

• 한국산학기술학회논문지
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• 제15권11호
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• pp.6774-6781
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• 2014

인삼(Panax ginseng C. A. Meyer)의 주요 생리활성물질인 진세노사이드(ginsenoside) Rb1과 Rg1의 효능검증 및 작용점을 규명하고자 HaCaT 피부각질세포에서 유전체 분석(gene expression profiles)을 실시하였다. 진세노사이드 Rb1과 Rg1 각각의 처리 농도 및 시간에 따른 HaCaT 세포에 대한 세포독성은 나타나지 않았으며, $10{\mu}g/mL$의 진세노사이드 Rb1과 Rg1 각각을 6 및 24 시간 처리하여 유전체 분석 결과, 진세노사이드 Rb1과 Rg1의 24 시간 처리군에서 항노화 및 피부탄력 관련 유전자인 fibroblast growth factor (FGF2)의 활성이 증가된 것으로 나타났다. 또한 진세노사이드 Rb1의 24 시간 처리군에서는 항산화 작용점에 있는 일련의 유전자군, FANCD2, FGF2, LEPR, FAS 등의 활성을 확인하였다. 향후 확인된 항노화 및 피부탄력 관련 주요인자들의 작용 및 상관관계를 구체적으로 확인하고, 특히 진세노사이드 Rb1의 신호전달을 완성하고자 한다.

• 대한한의학회지
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• 제44권2호
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• pp.119-131
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• 2023

Objectives: Betula Platyphylla(BP) has been used as a analgesic, anti-microbial, anti-oxidant drug in Eastern Asia. However, it is still unknown whether BP ethanol extract could exhibit the inhibitory activities against ultraviolet B(UVB)-induced skin injury on human keratinocytes, HaCaT cells. This study was aimed to investigate the protective activity of BP ethanol extract on UVB-irradiated skin injury in HaCaT cells. Methods: The skin injury model of HaCaT cells was established under UVB stimulation. HaCaT keratinocyte cells were pre-treated with BP ethanol extract for 1 h, and then stimulated with UVB. Then, the cells were harvested to measure the cell viability, production of reactive oxygen species(ROS), pro-inflammatory cytokines such as interleukin(IL) 1-beta, IL-6, and tumor necrosis factor(TNF)-𝛼, hyaluronidase, type 1 collagen, matrix metalloproteinase(MMP)s. In addition, we examined the mitogen activated protein kinases(MAPKs) and inhibitory kappa B alpha(I𝜅;-B𝛼) as inhibitory mechanisms of BP ethanol extract. Results: The treatment of BP ethanol extract inhibited the UVBinduced cell death and ROS production in HaCaT cells. BP ethanol extract treatment inhibited the UVB-induced increase of IL-1beta, IL-6, and TNF-𝛼. BP ethanol extract treatment inhibited the increase of hyaluronidase, MMP and decrease of collagen. BP ethanol extract treatment inhibited the activation of MAPKs and the degradation of I𝜅-B𝛼. Conclusions: Our result suggest that treatment of BP ethanol extract could inhibit the UVB-induced skin injury via deactivation of MAPKs and nuclear factor kappa B(NF-𝜅B) in HaCaT cells. This study could suggest that BP ethanol extract could be a beneficial agent to prevent skin damage or inflammation.