• Journal of Photoscience
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• 제9권2호
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• pp.500-502
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• 2002

In vitro and in vivo studies have reported the induction of matrix metaloproteinase (MMP)-1 in the fibroblasts by ultraviolet (UV) A irradiation. We constructed the skin equivalent model using HaCaT cells as keratinocytes and human neonatal dennal fibroblasts as fibroblasts in the present study. The induction of MMP-l in the fibroblasts was confirmed immunohistochemically 6 hours after UVA irradiation using this model. This model was simply composed of human keratinocytes and fibroblasts. To our knowledge, there have been a few papers concerning the skin equivalent model in the field of photobiology. The effect of UVA exposure to fibroblasts through keratinocytes was examined using this model. The cross-talk can be examined between keratinocytes and fibroblasts. This model can be a useful tool in the field of photobiology.

• Biomolecules & Therapeutics
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• 제27권4호
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• pp.395-403
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• 2019

Purpurogallin, a natural phenol obtained from oak nutgalls, has been shown to possess antioxidant, anticancer, and anti-inflammatory effects. Recently, in addition to ultraviolet B (UVB) radiation that induces cell apoptosis via oxidative stress, particulate matter 2.5 ($PM_{2.5}$) was shown to trigger excessive production of reactive oxygen species. In this study, we observed that UVB radiation and $PM_{2.5}$ severely damaged human HaCaT keratinocytes, disrupting cellular DNA, lipids, and proteins and causing mitochondrial depolarization. Purpurogallin protected HaCaT cells from apoptosis induced by UVB radiation and/or $PM_{2.5}$. Furthermore, purpurogallin effectively modulates the pro-apoptotic and anti-apoptotic proteins under UVB irradiation via caspase signaling pathways. Additionally, purpurogallin reduced apoptosis via MAPK signaling pathways, as demonstrated using MAPK-p38, ERK, and JNK inhibitors. These results indicate that purpurogallin possesses antioxidant effects and protects cells from damage and apoptosis induced by UVB radiation and $PM_{2.5}$.

• Journal of Cancer Prevention
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• 제21권3호
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• pp.135-143
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• 2016

Background: Fraxetin (7,8-dihydroxy-6-methoxy coumarin), a coumarin derivative, has been reported to possess antioxidative, anti-inflammatory and neuroprotective effects. A number of recent observations suggest that the induction of heme oxygenase-1 (HO-1) inhibits inflammation and tumorigenesis. In the present study, we determined the effect of fraxetin on HO-1 expression in HaCaT human keratinocytes and investigated its underlying molecular mechanisms. Methods: Reverse transcriptase-PCR and Western blot analysis were performed to detect HO-1 mRNA and protein expression, respectively. Cell viability was measured by the MTS test. The induction of intracellular reactive oxygen species (ROS) by fraxetin was evaluated by 2′,7′-dichlorofluorescin diacetate staining. Results: Fraxetin upregulated mRNA and protein expression of HO-1. Incubation with fraxetin induced the localization of nuclear factor-erythroid-2-related factor-2 (Nrf2) in the nucleus and increased the antioxidant response element-reporter gene activity. Fraxetin also induced the phosphorylation of Akt and AMP-activated protein kinase $(AMPK){\alpha}$ and diminished the expression of phosphatase and tensin homolog, a negative regulator of Akt. Pharmacological inhibition of Akt and $AMPK{\alpha}$ abrogated fraxetin-induced expression of HO-1 and nuclear localization of Nrf2. Furthermore, fraxetin generated ROS in a concentration-dependent manner. Conclusions: Fraxetin induces HO-1 expression through activation of Akt/Nrf2 or $AMPK{\alpha}/Nrf2$ pathway in HaCaT cells.

• 대한의생명과학회지
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• 제25권1호
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• pp.15-22
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• 2019

The diagnosis of atopic dermatitis (AD) includes a test that checks allergen-mediated skin reactions and a method of measuring the total IgE and allergen-specific IgE in blood. However, these test methods are performed directly on the patient, which cause some pain or discomfort. In addition, the skin response test or IgE may result in false negative in about 20% of patients. In the present study, to identify specific biomarkers, HaCaT cells were used as a human keratinocyte that make up the skin, were treated IL-4 and IL-13 for 24 hours to induce a situation similar to keratinocytes in AD patients. In the HaCaT cells, pro-inflammatory cytokine such as IL-5, IL-6, and MCP-1 were increased by IL-4 and IL-13 and skin barrier proteins was reduced by IL-4 and L-13. This results showed that a situation similar to the stratum corneum of an actual patient is induced in HaCaT cells. And then the secretions of Kallikrein (KLK) 5 and KLK7 protease were checked by enzyme-linked immunosorbent assay (ELISA). It was specifically increased by IL-4 and IL-13. This showed that AD-related protease can be detected at the protein level using keratinocytes that can be taken in a non-invasive manner and suggested the possibility of applying it to AD diagnosis.

• 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.

• 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 Applied Biological Chemistry
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• 제61권4호
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• pp.417-421
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• 2018

자외선은 가시 광선보다 파장이 짧은 전자기파이며, 오존층을 통과하는 자외선이 피부 노화의 주요 원인이다. 피부 조직을 자외선에 만성적으로 노출 시키면 인간 각질 형성 세포에서 Mitogen-activated Protein Kinases (MAPKs) 신호 전달 경로가 활성화되어 매트릭스 metalloproteinases (MMPs) 생성이 증가한다. 본 연구에서는 제주도의 초본 추출물을 대상으로 자외선 자극을 통해 인간 각질 형성 세포(HaCaTs)의 항 노화 효과에 대해 조사 하였다. 우리는 자외선 유도 된 각질 형성 세포에서 MMP-1 유전자와 단백질 발현 수준을 측정 하였으며 그 결과, Thymus quinquecostatus 추출물(TQE)은 자외선 조사된 HaCaT 세포에서 의해 농도 의존적으로 MMP-1의 발현을 유의하게 감소시키는 것으로 확인되었다. 또한, TQE는 MAPK 신호 전달요소 인 ERK1/2, JNK1/2 및 p38 단백질의 자외선 유도에 따른 인산화를 유의 적으로 감소시켰다. 이러한 결과는 MAPKs 경로가 인간 각질 세포에서 UV 유도 MMP-1 생산에 대한 TQE의 억제 효과에 기여할 수 있음을 시사한다. 우리의 결과는 아마도 인간 각질형성 세포에서 TQE가 자외선 유도된 MMP-1 생산을 억제하는 피부 노화 소재로 활용될 수 있음을 시사한다.

• 한방안이비인후피부과학회지
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• 제20권3호
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• pp.14-28
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• 2007

Objective : Hwagi-Jokyeong-Tang (化氣調經湯, HJT) was described in DongeuiBogam(東醫寶鑑). This remedy has been used to treat patients with Naryeok, which is similar as tuberculous cervical lymphadenitis in western medicine. Methods : In this study, the present author investigated the effects of HJT on on Human HaCaT keratinocyte and malignant melanoma cells such as SK-MEL-2 and B16F10 in terms of cell viabilities, proliferations, DPPH free radical scavenging activities, oxygen free radical productions and inhibitory action on elastase activities. Results : HJT acceleated proliferation of HaCaT keratinocytes dose-dependantly. HJT also prevented cell death of HaCaT induced by Hydrogen peroxide, which products oxygen free radicals. On the contrary, HJT did not affect proliferations of SK-MEL-2 or B16F10. In addition, HJT was shown to have DPPH free radical scavenging activities and also have inhibitory effects on elastase activities too. On the fluorescent examinations, the present author know that HJT did not affect production levels of oxygen free radicals in malignant melanoma cell, SK-MEL-2. Conclusions : These results suggest that HJT has possibilities of usage for functional cosmetics which have skin regeneration or prevention from skin tissue injury.

• Biomolecules & Therapeutics
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• 제29권1호
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• pp.90-97
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• 2021

Ultraviolet B (UVB) radiation causes DNA base modifications. One of these changes leads to the generation of 8-oxoguanine (8-oxoG) due to oxidative stress. In human skin, this modification may induce sunburn, inflammation, and aging and may ultimately result in cancer. We investigated whether phloroglucinol (1,3,5-trihydroxybenzene), by enhancing the expression and activity of 8-oxoG DNA glycosylase 1 (Ogg1), had an effect on the capacity of UVB-exposed human HaCaT keratinocytes to repair oxidative DNA damage. Here, the effects of phloroglucinol were investigated using a luciferase activity assay, reverse transcription-polymerase chain reactions, western blot analysis, and a chromatin immunoprecipitation assay. Phloroglucinol restored Ogg1 activity and decreased the formation of 8-oxoG in UVB-exposed cells. Moreover, phloroglucinol increased Ogg1 transcription and protein expression, counteracting the UVB-induced reduction in Ogg1 levels. Phloroglucinol also enhanced the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) as well as Nrf2 binding to an antioxidant response element located in the Ogg1 gene promoter. UVB exposure inhibited the phosphorylation of protein kinase B (PKB or Akt) and extracellular signal-regulated kinase (Erk), two major enzymes involved in cell protection against oxidative stress, regulating the activity of Nrf2. Akt and Erk phosphorylation was restored by phloroglucinol in the UVB-exposed keratinocytes. These results indicated that phloroglucinol attenuated UVB-induced 8-oxoG formation in keratinocytes via an Akt/Erk-dependent, Nrf2/Ogg1-mediated signaling pathway.