Effects of itraconazole and tacrolimus on Aryl hydrocarbon receptor and NADPH quinine oxidoreductase 1

( Jee Hee Son ),( Sook Young Park ),( Yong Se Cho ),( Yun Sun Byun ),( Yoon Seok Yang ),( Bo Young Chung ),( Hee Jin Cho ),( Hye One Kim ),( Chun Wook Park ) 대한피부과학회 2015 대한피부과학회 학술발표대회집 Vol.67 No.2

Background: Aryl hydrocarbon receptor (AhR) is a ligand-inducible transcription factor responding to halogenated aromatic hydrocarbons. AhR regulates nuclear factor-erythroid 2- related factor-2 (Nrf2), a key molecule for cell protection. Itraconazole (ICZ) and tacrolimus exhibit anti-inflammatory effects, but underlying molecular mechanism remains unclear, especially on AhR-Nrf2 pathway. Objectives: To evaluate the influence of ICZ and tacrolimus on AhR pathway, especially AhR-Nrf2 pathway. Methods: Normal human epidermal keratinocytes (NHEK) were treated with 2,3,7,8-tetrachlrodibenzo-p-dioxin TCDD), 6-Formylindolo[3,2-b]carbazole (FICZ), Indirubin, ICZ and tacrolimus. Then, RT-PCR and Western blot analysis were performed. Results: The expression levels of the AhR, Arylhydrocarbon receptor nuclear translocator (ARNT), Cytochrome P450 1A1 (CYP1A1) and NADPH quinine oxidoreductase 1 (NQO1) mRNA were higher when treated with TCDD, FICZ, and Indirubin, especially in 12 hours. However, with ICZ and tacrolimus, there was no significant difference from control. When analyzing protein expressions of NHEKs with Western blot, the levels of AhR, CYP1A1, and NQO1 were higher after treatment with TCDD, FICZ, and Indirubin. With ICZ and tacrolilmus, there was no significant difference Conclusion: ICZ and tacrolimus did not effect on mRNA and protein levels of AhR-related factors. This suggests ICZ and tacrolimus exhibit different pathway from AhR.

Benzo[a]pyrene Cytotoxicity Tolerance in Testicular Sertoli Cells Involves Aryl-hydrocarbon Receptor and Cytochrome P450 1A1 Expression Deficiencies

Kim, Jin-Tac,Park, Ji-Eun,Lee, Seung-Jin,Yu, Wook-Joon,Lee, Hye-Jeong,Kim, Jong-Min The Korean Society of Developmental Biology 2021 발생과 생식 Vol.25 No.1

Benzo[a]pyrene (B[a]P) is a potent carcinogen and is classified as an endocrine-disrupting chemical. In mammalian testes, Sertoli cells support spermatogenesis. Therefore, if these cells are negatively affected by exposure to xenotoxic chemicals, spermatogenesis can be seriously disrupted. In this context, we evaluated whether mouse testicular TM4 Sertoli cells are susceptible to the induction of cytotoxicity-mediated cell death after exposure to B[a] P in vitro. In the present study, while B[a]P and B[a]P-7,8-diol were not able to induce cell death, exposure to BPDE resulted in cell death. BPDE-induced cell death is accompanied by the activation of caspase-3 and caspase-7. Depolarization of the mitochondrial membrane and cytochrome c release from mitochondria were observed in benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated cells. These results indicate that TM4 cells are susceptible to apoptosis in a caspase-dependent manner. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that aryl hydrocarbon receptor (AhR) expression was almost undetectable in TM4 cells and that its expression was not altered after B[a]P treatment. This indicates that TM4 cells are nearly AhR-deficient. In TM4 cells, the CYP1A1 protein and its activity were not present. From these results, it is clear that AhR may be a prerequisite for CYP1A1 expression in TM4 cells. Therefore, TM4 cells can be referred to as CYP1A1-deficient cells. Thus, TM4 Sertoli cells are believed to have a rigid and protective cellular machinery against genotoxic agents. In conclusion, it is suggested that tolerance to B[a]P cytotoxicity is associated with insufficient AhR and CYP1A1 expression in testicular Sertoli cells.

2,3,7,8-Tetrachlorodibenzo-p-Dioxin Regulates the Expression of Aryl Hydrocarbon Receptor-Related Factors and Cytokines in Peripheral Blood Mononuclear Cells and CD4+ T cells from Patients with Atopic Dermatitis and Psoriasis

엄지영,김한비,강석영,손지희,정보영,박천욱,김혜원 대한피부과학회 2020 Annals of Dermatology Vol.32 No.5

Background: Aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, is important for xenobiotic metabolism and binds to various endogenous and exogenous ligands in the skin. However, the functional role of AhR in patients with psoriasis (PS) and atopic dermatitis (AD) remains unclear. Objective: We aimed to determine whether AhR-regulated factors (AhR, CYP1A1, interleukin [IL]-17, IL-22) were affected by AhR ligands (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) in chronic inflammatory skin diseases such as PS and AD. Methods: The expression levels of AhR-related factors were determined by quantitative PCR, western blotting, and immunocytochemistry. Specific siRNA targeting AhR was used to inhibit gene expression in human peripheral blood mononuclear cells (PBMC). Cytokine assays were performed to determine the protein production of CD4+ T cells. Results: In comparison with healthy controls, TCDD-treated PBMCs and CD4+ T cells from patients with PS and AD showed an increase in AhR gene levels as well as significantly increased expression of AhR-related factors (such as AhR, CYP1A1, IL-17, and IL-22). In contrast, 6-formyl indolo [3,2-b] carbazole (FICZ) inversely affected the differentiation of CD4+ T cells and their cytokine expression levels as compared with TCDD. CD4+ T cells from patients with AD and PS showed higher expression levels of AhR, CYP1A1, IL-17, and IL-22. Conclusion: Our results suggest that TCDD-induced AhR-related factor upregulation in AD and PS patients may increase the expression of AhR-regulatory genes, thereby contributing to the development of AD and PS.

Phenyldiazenylaniline 유도체가 방향족탄화수소 수용체의 활성에 미치는 영향

이효성 한국융합학회 2019 한국융합학회논문지 Vol.10 No.1

AHR regulates the expression of xenobiotics metabolizing enzymes (XMEs) as a transcription fact upon binding of ligands that are mainly aryl hydrocarbons. The role of AHR in human physiology has been intensively investigated for the past decades, however our understanding on AHR yet to be elucidated largely due to the lack of proper chemical agents. It has been demonstrated that AHR correlates to pathogenesis for some diseases in recent studies suggesting that the study on the AHR may provide a valid therapeutic target. Classical antagonists in current use are reported to be partially agonistic whereas a pure antagonist is yet to be found. In this study, phenyldiazenylaniline has been designed based on the structure of two known AHR antagonist, Resveratrol and CH223191. The derivatives of phenyldiazenylaniline have been prepared and subjected to assessment as an AHR antagonist in order to optimize the AHR antagonistic activity of the designed structure by means of convergence study of organic synthesis and molecular biology. 방향성 탄화수소 수용체(Aryl Hydrocarbon Receptor, AHR)은 리간드에 의해 활성화되어 체내 외래물질의 대사를 조절하는 전사인자다. 생체 내에서 AHR의 생리학적 역할은 오랜 기간 연구되어 왔으나 길항제를 비롯한 적절한 화학적 도구의 부재로 그 역할 규명이 제한되어 있다. AHR이 다양한 질병의 발병기전에 관여되어 있다는 것이 밝혀짐에 따라 유효한 약물 표적으로 인식되고 있으나 치료나 예방을 위한 유효한 약물은 아직 개발되지 않았다. 길항제로 알려진 화합물들은 낮은 농도에서는 길항활성이 있어 연구 목적으로는 활용되고 있으나 높은 농도에서는 AHR을 활성화하는 부분적 agonist로 작용한다. 이에 AHR 활성화를 유도하지 않는 순수한 길항제의 개발이 필요하다. 본 연구에서는 이미 알려진 AHR 길항제인 Resveratrol과 CH223191의 구조를 기반으로 phenyldiazenylanline 구조를 설계하였고 이를 골격으로 다양한 유도체를 합성하고 화학적 구조와 생물학적 활성의 상관 관계에 대한 융합 연구를 통하여 신규 AHR 길항제를 도출하였다.

Alpha-naphthoflavone induces apoptosis through endoplasmic reticulum stress via c-Src-, ROS-, MAPKs-, and arylhydrocarbon receptor-dependent pathways in HT22 hippocampal neuronal cells

Yu, Ah-Ran,Jeong, Yeon Ju,Hwang, Chi Yeon,Yoon, Kyung-Sik,Choe, Wonchae,Ha, Joohun,Kim, Sung Soo,Pak, Youngmi Kim,Yeo, Eui-Ju,Kang, Insug Elsevier 2019 NeuroToxicology Vol.71 No.-

<P><B>Abstract</B></P> <P>α-Naphthoflavone (αNF) is a prototype flavone, also known as a modulator of aryl hydrocarbon receptor (AhR). In the present study, we investigated the molecular mechanisms of αNF-induced cytotoxic effects in HT22 mouse hippocampal neuronal cells. αNF induced apoptotic cell death via activation of caspase-12 and -3 and increased expression of endoplasmic reticulum (ER) stress-associated proteins, including C/EBP homologous protein (CHOP). Inhibition of ER stress by treatment with the ER stress inhibitor, salubrinal, or by CHOP siRNA transfection reduced αNF-induced cell death. αNF activated mitogen-activated protein kinases (MAPKs), such as p38, JNK, and ERK, and inhibition of MAPKs reduced αNF-induced CHOP expression and cell death. αNF also induced accumulation of reactive oxygen species (ROS) and an antioxidant, N-acetylcysteine, reduced αNF-induced MAPK phosphorylation, CHOP expression, and cell death. Furthermore, αNF activated c-Src kinase, and inhibition of c-Src by a kinase inhibitor, SU6656, or siRNA transfection reduced αNF-induced ROS accumulation, MAPK activation, CHOP expression, and cell death. Inhibition of AhR by an AhR antagonist, CH223191, and siRNA transfection of AhR and AhR nuclear translocator reduced αNF-induced AhR-responsive luciferase activity, CHOP expression, and cell death. Finally, we found that inhibition of c-Src and MAPKs reduced αNF-induced transcriptional activity of AhR. Taken together, these findings suggest that αNF induces apoptosis through ER stress via c-Src-, ROS-, MAPKs-, and AhR-dependent pathways in HT22 cells.</P> <P><B>Highlights</B></P> <P> <UL> <LI> α-Naphthoflavone (αNF), an arylhydrocarbon receptor modulator (AhR), induces apoptosis in HT22 hippocampal neuronal cells. </LI> <LI> ER stress plays a role in αNF-induced apoptosis. </LI> <LI> The c-Src-, ROS-, MAPK-, and AhR-dependent pathways are implicated in αNF-induced ER stress and apoptosis. </LI> <LI> The AhR nuclear translocator-dependent genomic pathway also mediates αNF-induced ER stress and apoptosis. </LI> </UL> </P>

Air Pollution and Skin Physiology

( Hyun Jung Kim ) 한국피부장벽학회 2014 한국피부장벽학회지 Vol.16 No.2

Increased air pollution can be the critical health problem besides social and economic problem. Skin is the outermost organ of human body, so it must protect from various pollutants to maintain its homeostasis. In this review, the skin aging due to the exogenous factors, such as UV, ozone, smoking and air pollutant will be discussed related to aryl hydrocarbon receptor. Skin inflammatory disease- atopic dermatitis and psoriasis-cannot be indispensible from pollution and AhR. AhR modulating agents can be suggested as a new cosmetic and therapeutic target for new generation.

FC 1-3 : Increased expression of the aryl hydrocarbon receptor in patients with chronic inflammatory skin disease

( Ji Hye Kim ),( Yun Sun Byun ),( Yoon Seok Yang ),( Soo Ick Cho ),( Hye One Kim ),( Cheol Heon Lee ),( Chun Wook Park ) 대한피부과학회 2013 대한피부과학회 학술발표대회집 Vol.65 No.2

Background: Polychlorinated biphenyls (PCBs) and 2,3,7,8- tetrachlorodibenzo-p-dioxin (TCDD) are major environmental pollutants. The biological effects of TCDD and PCBs on the human body critically depend on the aryl hydrocarbon receptor (AhR). Objectives: The aim of this study was to evaluate the significance of the AhR and its ligands (TCDD and PCBs) in chronic inflammatory skin diseases such as AD and psoriasis. Methods: Immunohistochemistry (IHC), Immunofluorescence staining and real-time PCR was done in the lesional skin. Cytokine assay after treatment of normal human epidermal keratinocytes (NHEKs) with AhR ligands was done. Results: The AhR expression in the epidermis was significantly stronger in the lesional skin of AD and psoriasis compared to that of the control group as assessed by IHC. The AhR and aryl hydrocarbon receptor nuclear translocator (ARNT) were co-localized in the nuclei of keratinocytes at the lower epidermis of psoriatic lesions, which suggested activation of an AhR pathway. Expression of AhR-related mRNA was increased in lesional skin from AD and psoriasis patients compared to those of normal skin. After treatment of NHEKs with TCDD or PCBs, IL-6 and IL-8 production were increased. Conclusion: AhR is highly expressed in the acute lesional skin of AD and psoriasis patients and activation of the AhR increases the expression of inflammatory cytokines in NHEK and may induce the development of chronic inflammatory skin diseases such as AD and psoriasis.

Orginal Article : Increased Expression of the Aryl Hydrocarbon Receptor in Patients with Chronic Inflammatory Skin Disease

( Hye One Kim ),( Yun Sun Byun ),( Jin Hye Kim ),( Chun Wook Park ) 한국피부장벽학회 2014 한국피부장벽학회지 Vol.16 No.2

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that binds to various endogenous and exogenous ligands that are abundant in the skin. The aim of this study was to evaluate the significance of the AhR and its ligands in chronic inflammatory skin disease such as atopic dermatitis (AD) and psoriasis. We recruited AD, psoriasis patients and healthy controls. The clinical severity was assessed and skin samples were taken. Real-time PCR, Immunohistochemical and immunofluorescence staining were performed in each skin sample. Cytokine expression profiles after treatment of normal human epidermal keratinocytes (NHEK) with TCDD or PCBs were measured. The serum levels of TCDD and PCBs were determined using high-resolution gas chromatography and mass spectrometry and both levels were higher in the AD group than in the control group. In the skin, expression of AhR and aryl hydrocarbon receptor nuclear translocator (ARNT) mRNA was increased in lesional skin from patients with AD and psoriasis compared to those of normal skin from healthy controls. The AhR and ARNT were co-localized in the nuclei of keratinocytes at the lower epidermis of psoriatic lesions, which suggested activation of the AhR pathway. After treatment of NHEKs with TCDD or PCBs, IL-6 and IL-8 production were increased. The serum levels of TCDD and PCBs were higher in the AD group than in the control group. The results of this study suggest that the AhR is highly expressed in the acute lesional skin of AD and psoriasis patients and that activation of the AhR increases the expression of inflammatory cytokines in NHEK and may induce the development of chronic inflammatory skin diseases such as AD and psoriasis.

Aryl Sulfonamides Induce Degradation of Aryl Hydrocarbon Receptor Nuclear Translocator through CRL4DCAF15 E3 Ligase

김성아,조승현,조진화,Min Yeong Yu,신호철,김정애,Sung Goo Park,박병철,김선홍,김정훈 한국분자세포생물학회 2020 Molecules and cells Vol.43 No.11

Aryl hydrocarbon receptor nuclear translocator (ARNT) plays an essential role in maintaining cellular homeostasis in response to environmental stress. Under conditions of hypoxia or xenobiotic exposure, ARNT regulates the subset of genes involved in adaptive responses, by forming heterodimers with hypoxia-inducible transcription factors (HIF1α and HIF2α) or aryl hydrocarbon receptor (AhR). Here, we have shown that ARNT interacts with DDB1 and CUL4-associated factor 15 (DCAF15), and the aryl sulfonamides, indisulam and E7820, induce its proteasomal degradation through Cullin-RING finger ligase 4 containing DCAF15 (CRL4DCAF15) E3 ligase. Moreover, the two known neo-substrates of aryl sulfonamide, RNA-binding motif protein 39 (RBM39) and RNA-binding motif protein 23 (RBM23), are not required for ARNT degradation. In line with this finding, aryl sulfonamides inhibited the transcriptional activities of HIFs and AhR associated with ARNT. Our results collectively support novel regulatory roles of aryl sulfonamides in both hypoxic and xenobiotic responses.

P170 : Increased expression of the aryl hydrocarbon receptor in patients with chronic inflammatory skin disease

( Yun Sun Byun ),( Yong Se Cho ),( Yoon Seok Yang ),( Jin Hye Kim ),( Bo Young Chung ),( Chun Wook Park ),( Hye One Kim ) 대한피부과학회 2014 대한피부과학회 학술발표대회집 Vol.66 No.2

Background: Polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are major environmental pollutants, and their effects on the human body depend critically on the aryl hydrocarbon receptor (AhR). Objectives: The aim of this study was to evaluate the significance of the AhR and its ligands (TCDD and PCBs) in chronic inflammatory skin disease such as atopic dermatitis (AD) and psoriasis. Methods: We took the skin biopsy samples for real-time PCR and immunohistochemistry staining, and the blood samples for isolation of CD4+ T cell from AD, psoriasis patients and healthy controls. Differentiation of the cells was analyzed using FACS, ELISA, and real-time PCR. Results: The expression of AhR-related mRNA was increased in lesional skin from patients with AD and psoriasis. After treatment of normal human epidermal keratinocytes with TCDD or PCBs, IL-6 and IL-8 production were increased. In the CD4+ T cells with TCDD treatment, CCR10+ cells were significantly increased in psoriasis patients and CD161+CCR10+ cells (CCR10+: expressed in Th22 cells, CD161+: expressed in Th17 cells), levels of IL-4 were significantly increased in AD patients. In the TCDD-treatedgroup, levels of IFN-γ, IL-17, and IL-22 were significantly higher in psoriasis patients and levels of IL-13, IL-17, and IL-22 were significantly higher in AD patients compared to those of the healthy controls. Conclusion: AhR might be highly expressed in patients with AD and psoriasis.