title: In vitro effects of erythromycin on RANKL and nuclear factor-kappa B by human TNF-alpha stimulated Jurkat cells.
abstract: The receptor activator of NF-kappaB ligand (RANKL) and its signal downstream nuclear factor-kappaB (NF-kappaB) are critical regulators for immune responses as well as bone remodeling. The present study aimed to examine the effects of erythromycin (EM) on the activation of RANKL, correlation with NF-kappaB expression, proliferation and apoptosis of human Jurkat T cells. Jurkat T cells were pretreated with 100 pmol/l tumor necrosis factor-alpha (TNF-alpha) for 1 h followed by various concentrations of EM for 24 h. The mRNA expressions of RANKL and NF-kappaB were examined by RT-PCR. The protein expression of NF-kappaB was analyzed by Western blot. The protein level of RANKL was examined by flow cytometry, immunofluorescence microscopy and Western blot analyses. We also examined proliferation of Jurkat T cells by MTT assay, apoptosis by flow cytometry analysis after staining with PI and morphological observation after AO/EB staining. The results showed that EM inhibited TNF-alpha-induced expressions of RANKL and NF-kappaB at both mRNA and protein levels in a concentration-dependent manner. The expression of RANKL was correlated with the expression of NF-kappaB. Moreover, EM influenced the proliferation and apoptosis of human Jurkat T cells. These data suggest that EM acts as an anti-inflammatory agent not only to interact with the expression of NF-kappaB and the proliferation of human Jurkat T cells, but also to reduce the level of RANKL.
authors.affiliation: Department of Geratology, The First Affiliated Hospital of Chinese Medical University, Shenyang, China. wulan_2000@sina.com
authors.forename: Lan
authors.initials: L
authors.lastname: Wu
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Lin
authors.forename: Kai
authors.initials: K
authors.lastname: Bao
authors.forename: Peng-Fei
authors.initials: PF
authors.lastname: Li
authors.forename: Wei-Ge
authors.initials: WG
authors.lastname: Zhang
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004917:Erythromycin / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D053245:RANK Ligand / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D015533:Transcriptional Activation / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D016328:NF-kappa B; D053245:RANK Ligand; C505688:TNFSF11 protein, human; D014409:Tumor Necrosis Factor-alpha; D004917:Erythromycin