• 대한한방부인과학회지
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• 제24권4호
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• pp.31-49
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• 2011

Objectives: This study was performed to assess the protective effect of Polygonum multiflorum(PM) on UVB-irradiated HaCaT Keratinocytes damage. Methods: The protective effects of Polygonum multiflorum(PM) were determined by UVB-irradiated HaCaT assay. We assessed protective effects of Polygonum multiflorum(PM) on LDH release and nitrite production from HaCaT. COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-${\kappa}B$, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. PM inhibited LDH Release in UVB-irradiated HaCaT Keratinocytes. 2. PM inhibited Nitrite Production in UVB-irradiated HaCaT Keratinocytes. 3. PM suppressed the Gene Expression of COX-2 in UVB-irradiated HaCaT Keratinocytes. 4. PM increased the Gene Expression of Bcl-2 in UVB-irradiated HaCaT Keratinocytes. 5. PM didn't increase the Gene Expression of Bax in UVB-irradiated HaCaT Keratinocytes. 6. PM suppressed the Gene Expression of $TNF{\alpha}$ in UVB-irradiated HaCaT Keratinocytes. 7. PM suppressed the Gene Expression of c-jun in UVB-irradiated HaCaT Keratinocytes. 8. PM suppressed the Gene Expression of c-fos in UVB-irradiated HaCaT Keratinocytes. 9. PM suppressed the Gene Expression of NF-${\kappa}B$ in UVB-irradiated HaCaT Keratinocytes. 10. PM suppressed the Gene Expression of i-NOS in UVB-irradiated HaCaT Keratinocytes. 11. PM didn't increase the Gene Expression of Bcl-xL in UVB-irradiated HaCaT Keratinocytes Conclusions: In conclusion, these results suggest that PM inhibited the cell damage in UVB-irradiated HaCaT.

• 대한한방부인과학회지
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• 제24권2호
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• pp.33-51
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• 2011

Objectives: This study was performed to assess the protective effect of Saengmaek-san (SM) on UVB-induced HaCaT cell damage. Methods: The protective effects of Saengmaek-san(SM) were determined by UVB-induced HaCaT assay. We assessed protective effects of Saengmaek-san (SM) on LDH release and nitrite release from HaCaT. And COX-2, Bcl-2, Bax, $TNF{\alpha}$, c-jun, c-fos, NF-kB, iNOS, Bcl-xL gene expression were determined in HaCaT using real-time PCR method. Results: 1. SM inhibited LDH-release, nitrite production in UVB-exposed HaCaT. 2. SM suppressed the gene expression of COX-2, $TNF{\alpha}$ in UVB-exposed HaCaT. 3. SM increased the gene expression of Bcl-2, Bax, Bcl-xL family protein in UVB-exposed HaCaT. 4. SM suppressed the gene expression of c-jun, c-fos, NF-kB in UVB-exposed HaCaT. Conclusions: The study showed SM inhibited the cell damage in UVB-exposed HaCaT.

• 대한본초학회지
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• 제28권4호
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• pp.57-62
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• 2013

Objectives : Lonicerae Japonicae Flos (LJF) has been shown anti-oxidant, anti-inflammatory, anti-viral, anti-rheumatoid properties. However, it is still largely unknown whether LJF inhibits skin injury against oxidative stress in human keratinocyte, HaCaT cells. The purpose of this study was to evaluate the protective effects of LJF against hydrogen peroxide($H_2O_2$)-induced oxidative stress in human keratinocytes, HaCaT cells. Methods : To evaluate out the protective effects of LJF on oxidative injury in HaCaT cells, an oxidative stress model of HaCaT cells was established under a suitable concentration (500 ${\mu}M$) hydrogen peroxide. HaCaT keratinocyte cells were pre-treated with LJF (0.1, 0.25 or 0.5 mg/ml), and then stimulated with $H_2O_2$. Then, the cells were harvested to measure the cell viability, DNA damage, and release of reactive oxygen species (ROS). Results : LJF (0.1, 0.25 or 0.5 mg/ml) itself did not show any significant toxicity in HaCaT cells. The treatment of $H_2O_2$ caused the oxidative stress, leading to the cell death, and DNA injury. However, pretreatment with LJF reduced cell death, and DNA injury. The stimulation of $H_2O_2$ on HaCaT cells resulted in excessive release of ROS, which is the main factor of oxidative stress. The excessive release of ROS was inhibited by LJF treatment significantly. Conclusions : These results could suggest that LJF exhibited the protective effects of HaCaT cells against $H_2O_2$-induced oxidative stress by inhibiting ROS release. It could be explained that LJF inhibit skin damages against oxidative stress. Thus, LJF would be useful for the development of drug or cosmetics treating skin troubles.

• Biomolecules & Therapeutics
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• 제20권2호
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• pp.171-176
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• 2012

HaCaT cells are the immortalized human keratinocytes and have been extensively used to study the epidermal homeostasis and its pathophysiology. T helper cells play a role in various chronic dermatological conditions and they can affect skin barrier homeostasis. To evaluate whether HaCaT cells can be used as a model cell system to study abnormal skin barrier development in various dermatologic diseases, we analyzed the gene expression profile of epidermal differentiation markers of HaCaT cells in response to major T helper (Th) cell cytokines, such as $IFN{\gamma}$, IL-4, IL-17A and IL-22. The gene transcriptional profile of cornified envelope-associated proteins, such as filaggrin, loricrin, involucrin and keratin 10 (KRT10), in HaCaT cells was generally different from that in normal human keratinocytes (NHKs). This suggests that HaCaT cells have a limitation as a model system to study the pathophysiological mechanism associated with the Th cell cytokine-dependent changes in cornified envelope-associated proteins which are essential for normal skin barrier development. In contrast, the gene transcription profile change of human ${\beta}2$-defensin (HBD2) in response to $IFN{\gamma}$, IL-4 or IL-17A in HaCaT cells was consistent with the expression pattern of NHKs. $IFN{\gamma}$ also up-regulated transglutaminase 2 (TGM2) gene transcription in both HaCaT cells and NHKs. As an alternative cell culture system for NHKs, HaCaT cells can be used to study molecular mechanisms associated with abnormal HBD2 and TGM2 expression in response to $IFN{\gamma}$, IL-4 or IL-17A.

• 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.

• 대한화장품학회지
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• 제44권4호
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• pp.375-379
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• 2018

본 연구진은 메틸말리올라이드가 인간 피부 세포주인 HaCaT 세포에서 NRF2를 유도하고 이를 통하여 항산화 효과를 나타내는지 알아보았다. MTT assay를 통하여 메틸말리올라이드는 $10{\mu}M$ 농도에서 24 h 동안 HaCaT 세포에 처리하여도 HaCaT 세포 독성을 나타내지 않는 것을 확인하였다. 본 연구진이 구축한 HaCaT-ARE-GFP-luciferase 세포에 메틸말리올라이드를 처리하고 루시퍼라아제 활성을 측정한 결과 메틸말리올라이드는 양성 대조군인 레스베라트롤보다 ARE 결합에 따른 루시퍼라아제 활성을 더욱 강하게 증가시키는 것을 확인하였다. 또한 HaCaT 세포에 메틸말리올라이드 처리 시 NRF2에 의하여 유도되는 항산화 단백질인 HO-1과 NQO1 mRNA 및 단백질 발현이 통계적으로 유의하게 증가하였다. 마지막으로 메틸말리올라이드는 HaCaT 세포에서 TPA에 의해서 유도된 DNA 및 지질의 산화를 강력하게 억제하였다. 결론적으로 본 연구는 메틸말리올라이드가 NRF2 유도를 통하여 피부 산화적 스트레스를 억제하며 이는 메틸말리올라이드가 신규 항산화 화장품의 소재로 적합하다는 것을 시사한다.

• 대한본초학회지
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• 제32권3호
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• pp.55-61
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• 2017

Objectives : The aim of this research was to determine the diverse effects of Lithospermi Radix Water Extract (LR) on human keratinocyte HaCaT cells, and to examine whether those effects could be applied to the human skin. Methods : We examined effect of LR on the cell viability of using the MTS assay in human keratinocyte HaCaT cells. The antioxidation effect of LR was analyzed relative to the well-known antioxidant resveratrol, using an ABTS assay. Quantitative RT-PCR analysis revealed that, in HaCaT cells, LR influenced the mRNA expression of tight-junction genes associated with skin moisturization. Furthermore, a wound-healing assay demonstrated altered cell migration in LR-treated HaCaT cells. Result : The cytotoxicity was confirmed to be higher in LR at a concentration of $800{\mu}g/m{\ell}$ using the MTS assay in HaCaT cells. In comparison to $100{\mu}M$ resveratrol, $1,600{\mu}g/m{\ell}$ LR showed either a similar or superior antioxidation effect. LR treatment in HaCaT cells reduced the mRNA expression levels of claudin 3, claudin 4, claudin 6, claudin 8, and ZO-2 to less than 0.80-fold, whereas JAM-A and Tricellulin mRNA expression level increased more than 1.33-fold. In addition, HaCaT cells migration was decreased to 83.9% by LR treatment. Conclusions : LR of antioxidation activity will have an anti-aging effect on the skin by reducing oxidative stress. Further studies are required to address the implications for human skin, given LR's effects of altering mRNA expression of tight junction-related gene and decreasing cell migration of HaCaT cells.

• 동의생리병리학회지
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• 제26권3호
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• pp.330-337
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• 2012

This study was performed to assess the Effect of SCF(Schisandrae Chinensis Fructus) on Keratinocyte Damage by UV irradiation. The effect of SCF were determined in UV irradiated HaCaT. We measured LDH release and NO release from HaCaT to elucidate the effect of SCF. And iNOS, TNF-${\alpha}$, COX-2, Bax, Bcl-2, Bcl-xL, c-jun, c-fos gene expression were determined in HaCat using real time PCR method. The results are as follows. SCF inhibited LDH-release, NO production in UV irradiated HaCaT. SCF increased the gene expression Bax, Bcl-2 and Bcl-xL protein in UV irradiated HaCaT. SCF suppressed the gene expression TNF-${\alpha}$ in UV irradiated HaCaT. SCF suppressed the gene expression iNOS, c-fos, and c-jun in UV irradiated HaCaT. SCF not affected the suppression of the gene expression COX-2 in UV irradiated HaCaT. The study showed SCF inhibited the cell damage in UV irradiated HaCaT.

• 한방안이비인후피부과학회지
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• 제31권2호
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• pp.11-23
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• 2018

목적 : 본 연구에서는 $TNF-{\alpha}$와 $IFN-{\gamma}$로 자극한 인간피부각질형성세포 (HaCaT keratinocytes) 모델을 사용하여 지모가 피부장벽 기능에 미치는 영향을 알아보고자 하였다. 방법 : MTT assay를 통하여 지모 주정(70% 에탄올) 추출물 (EAA)이 HaCaT keratinocytes의 세포생존율에 미치는 영향을 확인하였으며 wound healing assay를 통해 EAA가 HaCaT 세포의 이주 능력에 영향을 주는지 관찰하였다. 또한 western blot analysis와 qRT-PCR을 통하여 EAA가 $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 발현 및 IL-4, IL-13, IL-6의 mRNA 발현, filaggrin의 단백질과 mRNA 발현에 미치는 영향을 조사하였다. 결과 : EAA는 처리 농도 $500{\mu}g/ml$까지 HaCaT keratinocytes의 세포생존율에 영향을 미치지 않았다. EAA는 wound healing assay에서 HaCaT 세포의 이주 능력을 증가시켰으며, $TNF-{\alpha}/IFN-{\gamma}$로 자극한 HaCaT 세포에서 iNOS의 단백질 수준을 감소시켰다. 또한 EAA가 IL-4, IL-13, IL-6의 mRNA 발현을 억제하는 것 역시 확인할 수 있었다. 뿐만 아니라 EAA는 $TNF-{\alpha}/IFN-{\gamma}$ 자극에 의해 감소했던 filaggrin을 단백질과 mRNA 수준에서 회복시켰다. 결론 : EAA가 HaCaT 세포에서 Th2 type cytokines, pro-inflammatory cytokine의 억제와 filaggrin 회복을 통해 피부장벽 기능 손상에 대한 억제활성을 갖는 것을 확인하였으며, 이를 통해 EAA가 염증으로 인해 손상된 피부장벽 기능 개선에 효과적일 것으로 사료된다.

• 한국산학기술학회논문지
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• 제14권11호
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• pp.5778-5784
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• 2013

본 연구는 항주름 소재를 개발하기 위해서 목단피로부터 gallic acid (GA)를 분리하여 항산화능을 측정하였고, HaCaT 세포에서 세포독성을 측정하였다. 또한 HaCaT 세포에서 tumor necrosis factor alpha (TNF-${\alpha}$)에 의해 유도되는 matrix metalloproteinase-1 (MMP-1) mRNA 발현, protein 발현, 분비에 대한 GA의 영향을 관찰하였다. 결과로써 GA는 30 ${\mu}g/mL$의 $IC_{50}$과 함께 항산화능을 나타내었고, 그것의 항산화능은 합성항산화제인 butylated hydroxyanisol (BHA)보다 높았다. GA는 HaCaT 세포에서 고농도인 200 ${\mu}g/mL$ 처리시 약한 세포독성을 나타내었다. 또한 HaCaT세포에서 TNF-${\alpha}$ (10 ng/mL)의 처리에 의해 증가된 MMP-1의 mRNA 발현, protein 발현, 분비는 GA의 처리에 의해 농도 의존적으로 유의적인 감소를 나타내었다(p<0.05). 그러므로 GA는 항산화 효과와 TNF-${\alpha}$로부터 유도되는 MMP-1의 발현을 저해함으로써 피부 주름을 개선할 수 있는 주름개선제로서의 활용가능성을 확인하였다.