• Archives of Pharmacal Research
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• 제26권9호
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• pp.739-746
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• 2003

Ultraviolet B (UVB) is known to induce apoptosis in human melanocytes. Here we show the cytoprotective effect of sphingosine-1-phosphate (S1P) against UVB-induced apoptosis. We also show that UVB-induced apoptosis of melanocytes is mediated by caspase-3 activation and poly(ADP-ribose) polymerase (PARP) cleavage, and that S1P prevents apoptosis by inhibiting this apoptotic pathway. We further investigated three major mitogen-activated protein (MAP) kinases after UVB irradiation. UVB gradually activated c-Jun N-terminal kinase (JNK) and p38 MAP kinase, while extracellular signal-regulated protein kinase (ERK) was inactivated transiently. Blocking of the p38 MAP kinase pathway using SB203580 promoted cell survival and inhibited the activation of caspase-3 and PARP cleavage. These results suggest that p38 MAP kinase activation may play an important role in the UVB-induced apoptosis of human melanocytes. To explain this cytoprotective effect, we next examined whether S1P could inhibit UVB-induced JNK and p38 MAP kinase activation. However, S1P was not found to have any influence on UVB-induced JNK or p38 MAP kinase activation. In contrast, S1P clearly stimulated the phosphorylation of ERK, and the specific inhibition of the ERK pathway using PD98059 abolished the cytoprotective effect of S1P. Based on these results, we conclude that the activation of p38 MAP kinase plays an important role in UVB-induced apoptosis, and that S1P may show its cytoprotective effect through ERK activation in human melanocytes.

• Biomolecules & Therapeutics
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• 제27권2호
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• pp.193-200
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• 2019

Ceramide metabolism is known to be an essential etiology for various diseases, such as atopic dermatitis and Gaucher disease. Glucosylceramide synthase (GCS) is a key enzyme for the synthesis of glucosylceramide (GlcCer), which is a main ceramide metabolism pathway in mammalian cells. In this article, we developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to determine GCS activity using synthetic non-natural sphingolipid C8-ceramide as a substrate. The reaction products, C8-GlcCer for GCS, could be separated on a C18 column by reverse-phase high-performance liquid chromatography (HPLC). Quantification was conducted using the multiple reaction monitoring (MRM) mode to monitor the precursor-to-product ion transitions of m/z $588.6{\rightarrow}264.4$ for C8-GlcCer at positive ionization mode. The calibration curve was established over the range of 0.625-160 ng/mL, and the correlation coefficient was larger than 0.999. This method was successfully applied to detect GCS in the human hepatocellular carcinoma cell line (HepG2 cells) and mouse peripheral blood mononuclear cells. We also evaluated the inhibition degree of a known GCS inhibitor 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) on GCS enzymatic activity and proved that this method could be successfully applied to GCS inhibitor screening of preventive and therapeutic drugs for ceramide metabolism diseases, such as atopic dermatitis and Gaucher disease.

• Animal Bioscience
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• 제34권1호
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• pp.143-153
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• 2021

Objective: To explore the molecular mechanisms of fatty liver hemorrhagic syndrome (FLHS) in laying hens, an experiment was conducted to reveal the differences in histopathological observation and gene expression between FLHS group and normal group. Methods: We compared the histopathological difference using hematoxylin and eosin staining and proceeded with RNA sequencing of adipose tissue to search differentially expressed genes and enriched biological processes and pathways. Then we validated the mRNA expression levels by real-time polymerase chain reaction and quantified protein levels in the circulation by enzyme-linked immunosorbent assay. Results: We identified 100 differentially expressed transcripts corresponding to 66 genes (DEGs) were identified between FLHS-affected group and normal group. Seven DEGs were significantly enriched in the immune response process and lipid metabolic process, including phospholipase A2 group V, WAP kunitz and netrin domain containing 2, delta 4-desaturase sphingolipid 2, perilipin 3, interleukin-6 (IL-6), ciliary neurotrophic factor (CNTF), and suppressor of cytokine signaling 3 (SOCS3). And these genes could be the targets of immune response and be involved in metabolic homeostasis during the process of FLHS in laying hens. Based on functional categories of the DEGs, we further proposed a model to explain the etiology and pathogenesis of FLHS. IL-6 and SOCS3 mediate inflammatory responses and the satiety hormone of leptin, induce dysfunction of Jak-STAT signaling pathway, leading to insulin resistance and lipid metabolic disorders. Conversely, CNTF may reduce tissue destruction during inflammatory attacks and confer protection from inflammation-induced insulin resistance in FLHS chickens. Conclusion: These findings highlight the therapeutic implications of targeting the JAK-STAT pathway. Inhibition of IL6 and SOCS3 and facilitation of CNTF could serve as a favorable strategy to enhance insulin action and improve glucose homoeostasis, which are of importance for treating obesity-related disorders for chickens.

• 대한화장품학회지
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• 제30권4호
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• pp.533-541
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• 2004

아토피 피부염은 다른 말로 태열이라고도 하며, 유아 시기에서부터 성인까지 광범위한 연령층에 병변의 특징적인 분포 그리고 개인적 혹은 가족적인 병력을 가진 특히 유전적 소인을 보이는 질환으로 심한 가려움증과 건조증 등을 동반하는 일종의 재발성, 만성의 알레르기성 습진을 말한다. 그 유병율은 매우 흔하여 어린이의 약 $9~12\%$ 에서발생한다. 그러나 최근에는 사회 자연환경의 변화와 음식문화의 변화, 삶의 방식의 변화에 의해서 계속 증가하는 추세를 보이고 있다. 인간의 피부는 외부 환경으로부터의 물리적, 화학적 자극에 대한 장벽기능의 역할을 하는데 아토피 피부염 환자의 피부는 각질층에서의 피부 장벽기능 및 수분유지 기능이 감소되어 있고 이러한 기능 장애의 원인으로 최근 연구에 의하면 세라마이드의 감소에 따른 것으로 밝혀지고 있다. 세라마이드는 스핑고신에 지방산이 연결되어 있는 구조를 가지고 있는 스핑고 지질의 일종이다. 세라마이드는 피부 각질층을 구성하는 각질세포간 지질 중 약 $40\%$를 차지하며, 수분 증발을 억제하는 지질 방어벽 역할과 각질층의 정연한 구조를 유지하게 하는 기능을 가지고 있다. 피부 각질층은 각화된 세포가 벽돌모양의 다층 구조로 구성되어 있으며 이러한 각화세포는 세라마이드, 콜레스테롤, 유리 지방산에 의해 견고히 결합되어 있다 따라서 이 세라마이드가 함유된 크림이나 연고를 아토피 피부염 환자에 도포함으로써 환자의 피부 장벽기능의 복구에 도움을 주어 아토피 피부염을 치료할 수 있다는 보고가 있다. 본 연구는 아토피 피부염 전용화장품으로서 아토피 피부에서 피부장벽을 복구하는데 기여한다고 증명된 각질 세포간 지질 성분들(세라마이드, 콜레스테롤, 유리지방산)을 포함하는 유액제품과 피부 면역조절과 항 염증 효과가 있다고 보고된 베타글루칸, 신이화 추출물 감초 추출물 등을 포함하는 세럼형태의 액상제품으로 구성된 두 가지 제품을 아토피 피부염 환자에게 도포하게 하여 그 제품의 안전성과 유효성을 확인하여 보았다. 그리고 그 제품이 새로운 시장인 아토피 피부염 전용 보습화장품으로서의 시장진입 가능성을 확인하여 보았다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1565-1574
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• 2018

Objective: The aim of the present study was to identify genetic variants based on RNA-seq data, obtained via transcriptome sequencing of muscle tissue of pigs differing in muscle histological structure, and to verify the variants' effect on histological microstructure and production traits in a larger pig population. Methods: RNA-seq data was used to identify the panel of single nucleotide polymorphisms (SNPs) significantly related with percentage and diameter of each fiber type (I, IIA, IIB). Detected polymorphisms were mapped to quantitative trait loci (QTLs) regions. Next, the association study was performed on 944 animals representing five breeds (Landrace, Large White, Pietrain, Duroc, and native Puławska breed) in order to evaluate the relationship of selected SNPs and histological characteristics, meat quality and carcasses traits. Results: Mapping of detected genetic variants to QTL regions showed that chromosome 14 was the most overrepresented with the identification of four QTLs related to percentage of fiber types I and IIA. The association study performed on a 293 longissimus muscle samples confirmed a significant positive effect of transforming acidic coiled-coil-containing protein 2 (TACC2) polymorphisms on fiber diameter, while SNP within forkhead box O1 (FOXO1) locus was associated with decrease of diameter of fiber types IIA and IIB. Moreover, subsequent general linear model analysis showed significant relationship of FOXO1, delta 4-desaturase, sphingolipid 1 (DEGS1), and troponin T2 (TNNT2) genes with loin 'eye' area, FOXO1 with loin weight, as well as FOXO1 and TACC2 with lean meat percentage. Furthermore, the intramuscular fat content was positively associated (p<0.01) with occurrence of polymorphisms within DEGS1, TNNT2 genes and negatively with occurrence of TACC2 polymorphism. Conclusion: This study's results indicate that the SNP calling analysis based on RNA-seq data can be used to search candidate genes and establish the genetic basis of phenotypic traits. The presented results can be used for future studies evaluating the use of selected SNPs as genetic markers related to muscle histological profile and production traits in pig breeding.

• 한국식품과학회지
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• 제44권4호
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• pp.434-440
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• 2012

쌀 유래 세라마이드를 함유한 미용보조제의 섭취에 관한 위약 대조, 이중 맹검 연구가 건조함으로 인해 항상 피부가 거친 경향이 있었던 33명의 피험자들에게 실시되어 다음과 같은 결과가 얻어졌다. 섭취 6주 후 피부과적 진단에 따르면 건조함, 홍조, 화장 유지력, 가려움 등의 관찰항목에서 플라시보 군 및 세라마이드군 모두에서 개선효과가 있었으나 플라시보 군 에서는 개선율이 43.8%인 반면, 세라마이드 군의 개선율은 64.7%로 높았다. 수분량, 유분량, pH 측정결과를 보면, 플라시보 군과 세라마이드 군 모두에서 유분량 및 pH 관련하여 유의한 변화는 없었다. 그러나 수분량에서는 플라시보 군에서 유의한 변화가 없었던 반면, 세라마이드 군에서는 6주 후 유의한 수분량 증가가 있었다. 3차원 현미경 경피분석시스템에 의한 분석결과에 따르면, 세라마이드 군에서는 Kurtosis, 유연성지수(SEsm), 거�s지수(SEr), 각질 및 인설 지수(SEsc), 주름지수(SEw)가 유의하게 개선되었으나 플라시보 군에서는 어떠한 유의한 변화도 관찰되지 않았다. 이는 이중맹검법으로 실시된 이번 연구에서 피부과적 진단이 명백한 플라시보 효과를 보인 점과 대비된다. 이로써 의사들에 의한 관찰과 더불어 과학적이고 객관적인 측정에 의해 세라마이드는 피부건조함과 거�s을 개선하는 데 효과가 있음이 입증되었고 경구섭취로 그 효과가 나타났다. 상기 결과는 쌀 유래 세라마이드 보조제의 장기간에 걸친 섭취가 보습, 피부의 유연성 유지에 효과적이고 따라서 효과적인 피부미용식품임을 보여준다.