title: Suppression of thymus- and activation-regulated chemokine (TARC/CCL17) production by 3-O-β-D-glucopyanosylspinasterol via blocking NF-κB and STAT1 signaling pathways in TNF-α and IFN-γ-induced HaCaT keratinocytes.
abstract: A phytosterol derivative, 3-O-β-D-glucopyanosylspinasterol (spinasterol-Glc) isolated from leaves of Stewartia koreana was reported to inhibit LPS-induced cytokine production in macrophage cells. Thymus and activation regulated chemokine (TARC/CCL17) is produced in response to pro-inflammatory cytokines in keratinocytes, which is implicated in the development of inflammatory skin diseases. In present study, we investigated the effect of spinasterol-Glc on production of TARC/CCL17 induced by TNF-α and IFN-γ in human HaCaT keratinocytes. Spinasterol-Glc inhibited the mRNA and protein expression of TARC/CCL17 induced by TNF-α/IFN-γ in a dose-dependent manner. Inhibitors of c-Raf-1, p38 MAPK, and JAK2, suppressed the TNF-α/IFN-γ-induced production of TARC/CCL17, and phosphorylation of these signaling molecules were attenuated by spinasterol-Glc. The compound also inhibited phosphorylation of IKKα/β and IκB-α, and reduced translocation of NF-κB to the nucleus. We demonstrated that spinasterol-Glc suppressed the NF-κB-driven and the GAS-driven expression of luciferase reporter gene induced by TNF-α and IFN-γ. In addition, spinasterol-Glc inhibited the DNA binding of NF-κB and STAT1 to its cognate binding site. These results suggest that spinasterol-Glc has effective inhibitory effects on production of TARC/CCL17 in keratinocytes via inhibition of NF-κB as well as STAT activation, and could be utilized for development of a potential therapeutic agent against skin inflammatory diseases.
authors.affiliation: Graduate School of Biotechnology and College of Life Sciences, Kyung Hee University, Yongin 446-701, Republic of Korea.
authors.forename: Mi-ra
authors.initials: MR
authors.lastname: Jung
authors.forename: Tae Hoon
authors.initials: TH
authors.lastname: Lee
authors.forename: Myun-Ho
authors.initials: MH
authors.lastname: Bang
authors.forename: Hakwon
authors.initials: H
authors.lastname: Kim
authors.forename: Youngsook
authors.initials: Y
authors.lastname: Son
authors.forename: Dae Kyun
authors.initials: DK
authors.lastname: Chung
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Kim
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002460:Cell Line; D054414:Chemokine CCL17 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D003876:Dermatitis, Atopic / Q000188:drug therapy / Q000378:metabolism; D003879:Dermatologic Agents / Q000494:pharmacology*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D053614:Janus Kinase 2 / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D010766:Phosphorylation / Q000187:drug effects; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000037:antagonists & inhibitors*; D015398:Signal Transduction / Q000187:drug effects; D013265:Stigmasterol / Q000031:analogs & derivatives* / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: C504840:3-O-beta-D-glucopyranosyl-spinasterol; D000894:Anti-Inflammatory Agents, Non-Steroidal; C106230:CCL17 protein, human; D054414:Chemokine CCL17; D003879:Dermatologic Agents; D016328:NF-kappa B; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D013265:Stigmasterol; C507924:JAK2 protein, human; D053614:Janus Kinase 2; D019908:Proto-Oncogene Proteins c-raf; D048051:p38 Mitogen-Activated Protein Kinases