• 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.

• 운동영양학회지
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• 제24권4호
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• pp.7-14
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• 2020

[Purpose] This study evaluated the anti-atopic dermatitis (AD) properties of Cordyceps militaris (CM) aqueous extract in keratinocytes in vitro and in vivo. We investigated the nutraceutical composition of the CM extract, including its protein, carbohydrate, and selected phytochemical content. [Methods] The expression of pathogenic cytokines in keratinocytes was assayed using an in vitro model. The CM extract downregulated extracellular signalregulated kinase 1/2 (ERK1/2) and p38 kinase expression in TNFα/IFNγ-stimulated HaCaT cells. We also established an in vivo AD model by repeatedly exposing the ears of mice to local Dermatophagoides farinae extract (DFE; house dust mite extract) and 2,4-dinitrochlorobenzene (DNCB). The epidermal and dermal ear thickness, mast cell infiltration, and serum immunoglobulin levels were measured following a 4-week oral administration of the CM extract. [Results] Histopathological examination showed reduced epidermal/dermal thickness and mast cell infiltration in mouse ears. The CM extract also suppressed serum immunoglobulin levels and gene expression of T helper (Th)1/Th2 cytokines in mouse ear tissue. [Conclusion] These results suggest that the CM extract may be useful for the treatment of AD-like skin lesions.

• Food Science and Biotechnology
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• 제18권2호
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• pp.475-480
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• 2009

Ultraviolet B (UVB) radiation provokes the generation of reactive oxygen species (ROS) in the cells and skin, which induce oxidative stress in the exposed cells, leading to photoaging and cancer. Using the human keratinocytes HaCaT cell line, we investigated the photoprotective effects of aucubin isolated from Eucommia ulmoides. Pretreatment with aucubin markedly suppressed UVB-induced oxidative stress, which manifests as a decrease in intracellular lipid peroxidation, elevation of catalase activity, and reduced glutathione content. In addition, aucubin significantly reduced expression of matrix metalloproteinase-1 (MMP-1) protein (54%) and mRNA. Taken together, these results suggest that aucubin may offer protection against UVB-induced oxidative stress and may be used as a potential agent in prevention of UVB-induced photoaging.

• 대한약학회:학술대회논문집
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• 대한약학회 2004년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.103-103
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• 2004

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.182.2-183
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• 2001

• 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.

• Journal of Applied Biological Chemistry
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• 제65권4호
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• pp.405-412
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• 2022

Echinodorus cordifolius (L.)은 택사과(Alismataceae)에 속하는 수생 식물이다. Echinodorus cordifolius (L.)의 피부 항노화 활성에 대한 연구는 아직까지 보고되지 않았다. 따라서 본 연구의 목적은 Echinodorus cordifolius (L.)로부터 70% 에탄올 추출물 (ECEE)을 제조하고 항산화 및 hyaluronidase 억제 활성을 통해 피부 항노화 기능성 소재로서의 가능성을 확인하였다. ECEE는 EC₅₀과 IC₅₀ 값이 각각 31.4, 300, 450 ㎍/mL로 우수한 DPPH, hydrogen peroxide 소거 활성 및 hyaluronidase 억제 활성을 가지는 것으로 나타났다. ECEE는 불멸화된 인간 각질세포인 HaCaT 세포에서 1 mM H₂O₂로 유도된 산화적 스트레스에 대해 농도 의존적으로 세포 보호 효과 증가 및 세포 내 활성산소종을 생성을 억제하는 것으로 나타났다. 또한 ECEE는 hyaluronic acid (HA) 합성 효소인 hyaluronan synthase 2 발현 수준을 상향 조절하고 HA 분해 효소인 hyaluronidase 2(HYAL2)의 발현 수준을 하향 조절하여 HaCaT 세포에서 HA 생성량을 증가시켰다. 따라서 이러한 결과를 종합하면 ECEE는 항산화, hyaluronidase 억제 및 HA 생성 촉진의 우수한 잠재력을 가진다는 것을 확인하였고 이를 통해 피부 노화를 방지하기 위한 천연 기능성 화장품 소재로서의 충분한 활용 가치가 있다고 판단된다.

• Journal of Microbiology and Biotechnology
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• 제32권11호
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• pp.1390-1395
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• 2022

Acne is a chronic inflammatory disease of the sebaceous gland attached to the hair follicles. Cutibacterium acnes is a major cause of inflammation caused by acne. It is well known that C. acnes secretes a lipolytic enzyme to break down lipids in sebum, and free fatty acids produced at this time accelerate the inflammatory reaction. There are several drugs used to treat acne; however, each one has various side effects. According to previous studies, sulforaphene (SFEN) has several functions associated with lipid metabolism, brain function, and antibacterial and anti-inflammatory activities. In this study, we examined the effects of SFEN on bacterial growth and inflammatory cytokine production induced by C. acnes. The results revealed that SFEN reduced the growth of C. acnes and inhibited proinflammatory cytokines in C. acnes-treated HaCaT keratinocytes through inhibiting NF-κB-related pathways. In addition, SFEN regulated the expression level of IL-1α, a representative pro-inflammatory cytokine expressed in co-cultured HaCaT keratinocytes and THP-1 monocytes induced by C. acnes. In conclusion, SFEN showed antibacterial activity against C. acnes and controlled the inflammatory response on keratinocytes and monocytes. This finding means that SFEN has potential as both a cosmetic material for acne prevention and a pharmaceutical material for acne treatment.