• 동의생리병리학회지
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• 제32권4호
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• pp.226-231
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• 2018

Chijabaegpi-tang (CHG) is an oriental herbal medicine that has been used for its various pharmacological effects, which include anti-inflammatory, anti-oxidant and immunoregulation activities. In the present study, we investigated which skin inflammations are involved in the $TNF-{\alpha}/IFN{\gamma}$-induced HaCaT cells. We investigated the suppressive effect of CHG on $TNF-{\alpha}/IFN{\gamma}$-induced HaCaT cell production of the following chemokines: macrophage-derived chemokine (MDC)/CCL22; regulated on activation, normal T-cell expressed and secreted (RANTES)/CCL5; and interleukin-8 (IL-8); thymus and activation-regulated chemokine (TARC)/CCL17. The pre-treatment of HaCaT cells with CHG suppressed $TNF-{\alpha}/IFN{\gamma}$-induced nuclear transcription factor kappa-B ($NF-{\kappa}B$). In addition, CHG inhibited $TNF-{\alpha}/IFN{\gamma}$-induced phosphorylation of ERK and p38. $TNF-{\alpha}/IFN{\gamma}$ suppressed the expression of skin barrier proteins, including filaggrin (FLG), Involucrin (IVL) and loricrin (LOR). By contrast, CHG restored the expression of FLG, IVL and LOR. Taken together, our findings suggest that CHG could be a therapeutic agent for prevention of skin disease, including atopic dermatitis.

• 대한임상검사과학회지
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• 제47권4호
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• pp.267-272
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• 2015

밀착연접(tight junction)은 사람의 표피에서 관찰되는 세포간 이음(intercellular junction) 중 하나로서, 표피 과립세포층의 세포간격에 위치하여 인접한 세포들을 접합시키고 전해질과 수분의 이동을 조절하는 장벽기능을 비롯한 다양한 생물학적 기능을 수행한다. 이러한 밀착연접의 대표적 단백성분인 occludin은 그 발현도가 밀착연접의 표피-생물학적 기능성과 높은 상관관계가 있는 것으로도 알려져 있다. 본 연구에서는 신체 여러 부위의 정상피부에서 occludin의 발현도 또는 발현정도의 상이성 여부를 알아보기 위하여 occludin에 대한 항체를 이용한 간접 면역형광검사를 시행하여 그 결과를 비교 검토하고자 하였다. 신체 7개 부위별로 각 4개씩의 정상 피부조직을 채취하여 항 occludin 항체를 이용한 간접 면역형광검사를 시행한 결과, 양성반응을 보이는 최종 희석배수의 역수(reciprocal of end-point titer)로 표시한 부위별 occludin 항원의 발현도는 두피에서 300, 얼굴에서 600, 목에서 300, 팔에서 200, 배에서 600, 그리고 등과 다리에서는 모두 400으로 관찰되어 신체 부위별 occludin의 발현도에는 통계학적으로 유의한 차이가 있음을 알 수 있었다(p=0.001). 본 연구에서 관찰된 신체 부위별 occludin의 발현도의 차이는 밀착연접의 부위별 발현의 차등을 시사하는 소견으로서, 표피 피부장벽의 생물학적 기능의 가능한 부위별 차등성과도 관계가 있을 것으로 사료된다.

• 대한화장품학회지
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• 제38권2호
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• pp.147-154
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• 2012

본 연구에서는 항산화 활성이 우수한 루틴을 담지한 탄성 리포좀을 제조하여, 이 제형의 물리적 특성과 증진된 피부 투과 효과를 평가하였다. 탄성 리포좀은 인지질과 계면활성제의 비율을 달리하여 제조하였다. 루틴을 담지한 탄성리포좀의 평균 입자 크기는 205.7 ~ 298.0 nm, 가변형성은 20.9 ~ 42.5, 포집효율은 52.0 ~ 71.0 %로 측정되었다. 0.1 % 루틴을 담지한 탄성 리포좀 중에서 인지질과 계면활성제 비율이 85 : 15 인 경우가 가장 높은 포집효율(71.0 %)과 가변형성 지수(42.5)를 나타내었다. 이 제형을 대상으로 피부 투과 실험을 진행하였다. 그 결과 대조군으로 사용된 일반 리포좀(98.0 ${\mu}g/cm^2$)과 1,3-butylene glycol (76.3 ${\mu} g/cm^2$) 용액보다 탄성 리포좀의 피부 투과능(129.9 ${\mu}g/cm^2$)이 훨씬 더 크게 나타났다. 이러한 결과들로 미루어 보아 $Tego^{(R)}$ care 450을 이용한 탄성 리포좀이 피부를 통한 유효성분 전달에 유용하게 이용될 수 있음을 확인하였다.

• 동의생리병리학회지
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• 제30권2호
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• pp.109-115
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• 2016

Hataedock is a Korean medical treatment that administers herbal extracts orally to newborn infants. This method is used for alleviating harmful heat and excreting fetal wastes by meconium. The purpose of this study was to evaluate anti-inflammatory effect of Hataedock method with Douchi on 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis (AD). The 3-week-old NC/Nga mice were divided into 3 groups: the control group (Ctrl), the AD-induced group (AE), and the Hataedock-treated group (GT). Only the GT group was treated with Hataedock at the 3rd week. After 28 days from Hataedock treatment, we induced AD-like dermatitis to the AE and GT group by DNFB. The effects of Hataedock were evaluated by immunohistochemical method. In the epithelium, PKC-positive reaction of the GT group was decreased by 57%. In the dermal papillae, IL-4-positive reaction was decreased by 34%. In the dermis, the distribution of degranulated mast cells was decreased and substance P-positive reaction was decreased by 49%. In the skin tissue, edema was decreased and MMP-9-positive reaction was decreased by 71%. Tissue damage such as epithelial cell hyperplasia, infiltration of granulocyte and lymphocyte, and capillary distribution were also decreased. The Hataedock method with Douchi maintained skin barrier and inhibited skin-damaging factors via regulating Th2 differentiation. In conclusion, Hataedock has a potential for preventative treatment of AD. Further studies are necessary to investigate the immune-regulating mechanism and verify the safety and efficacy of the Hataedock method.

• 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.

• Toxicological Research
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• 제32권2호
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• pp.109-114
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• 2016

Allergic skin inflammation such as atopic dermatitis is characterized by skin barrier dysfunction, edema, and infiltration with various inflammatory cells. The anti-inflammatory effects of Apo-9'-fucoxanthinone, isolated from Sargassum muticum, have been described in many diseases, but the mechanism by which it modulates the immune system is poorly understood. In this study, the ability of Apo-9'-fucoxanthinone to suppress allergic reactions was investigated using a mouse model of atopic dermatitis. The Apo-9'-fucoxanthinone-treated group showed significantly decreased immunoglobulin E in serum. Also, Apo-9'-fucoxanthinone treatment resulted in a smaller lymph node size with reduced the thickness and length compared to the induction group. In addition, Apo-9'-fucoxanthinone inhibited the expression of interleukin-4, interferon-gamma and tumor necrosis factor-alpha by phorbol 12-myristate 13-acetate and ionomycin-stimulated lymphocytes. These results suggest that Apo-9'-fucoxanthinone may be a useful therapeutic strategy for treating chronic inflammatory diseases.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.356-362
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• 2010

Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.192-209
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• 1995

The purpose of this study was to evaluate drug-loaded biodegradable membranes for guided tissue regeneration(GTR). The membranes were made by coating mesh of polyglycolic acid(PGA) with polylactic acid(PLA) containing 10% flurbiprofen or tetracycline. The thickness of membrane was $150{\pm}30{\mu}m$, and the pore size of surface was about $8{\mu}m$ in diameter. The release of drugs from the membrane was measured in vitro. Cytotoxity test for the membrane was performed by gingival fibroblast cell culture, and the tissue response was observed after implant of membrane into the dorsal skin of the rat for 8 wks. Ability to guided tissue regeneration of membranes were tested by measuring new bone in the calvarial defects(5mm in diameter) of the rat for 5 weeks. The amount of flurbiprofen and tetracycline released from membrane were about 30-60% during 7 days. Minimal cytotoxity was observed in the membrane except 20% drug containing membrane. In histologic finding of rat dorsal skin, many inflammatory cells were observed around e-PTFE, polyglactin 910 and PLAPGA membrane after 1 or 2 weeks. PLA-PGA membrane was perforated by connective tissue after 4 or 6 weeks, and divided as a segment at 8 weeks. In bone regeneration guiding potential test, tetracycline loaded membrane was most effective (p

• 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.

• 생약학회지
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• 제46권2호
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• pp.116-122
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• 2015

Keratinocytes are first barrier against outer challenges on skin. However, it is still largely unknown about effective protectors against ultraviolet B (UVB), and oxidative stress in human keratinocyte, HaCaT cells. Inducible heme oxygenase (HO)-1 acts against oxidants that are thought to play a role in the pathogenesis of skin disorders. Therefore, the purpose of this study was to evaluate the effect of Portulaca oleracea 70% EtOH extracts against hydrogen peroxide (H₂O₂)-induced oxidative stress in human keratinocytes, HaCaT cells. P. oleracea 70% EtOH extracts showed the potent protective effects on H₂O₂-induced toxicity by induced the expression of HO-1 in human keratinocyte, HaCaT cells. Furthermore, P. oleracea 70 % EtOH extracts caused the nuclear accumulation of nuclear factor E2-related factor 2 (Nrf2) in human keratinocytes, HaCaT cells. In addition, we found that treatment with c-Jun N-terminal kinase (JNK) inhibitor (SP600125) reduced P. oleracea 70% EtOH extracts-induced HO-1 expression, and JNK inhibitor (SP600125) also inhibited protective effects by P. oleracea 70% EtOH extracts. Therefore, these results suggest that P. oleracea 70 % EtOH extracts increases cellular resistance to H₂O₂-induced oxidative injury in human keratinocyte, HaCaT cells, presumably through JNK pathway-Nrf2-dependent HO-1 expression.