Azithromycin treatment alters gene expression in inflammatory, lipid metabolism, and cell cycle pathways in well-differentiated human airway epithelia.
Prolonged macrolide antibiotic therapy at low doses improves clinical outcome in patients affected with diffuse panbronchiolitis and cystic fibrosis. Consensus is building that the therapeutic effects are due to anti-inflammatory, rather than anti-microbial activities, but the mode of action is likely complex. To gain insights into how the macrolide azithromycin (AZT) modulates inflammatory responses in airways, well-differentiated primary cultures of human airway epithelia were exposed to AZT alone, an inflammatory stimulus consisting of soluble factors from cystic fibrosis airways, or AZT followed by the inflammatory stimulus. RNA microarrays were conducted to identify global and specific gene expression changes. Analysis of gene expression changes revealed that the AZT treatment alone altered the gene profile of the cells, primarily by significantly increasing the expression of lipid/cholesterol genes and decreasing the expression of cell cycle/mitosis genes. The increase in cholesterol biosynthetic genes was confirmed by increased filipin staining, an index of free cholesterol, after AZT treatment. AZT also affected genes with inflammatory annotations, but the effect was variable (both up- and down-regulation) and gene specific. AZT pretreatment prevented the up-regulation of some genes, such as MUC5AC and MMP9, triggered by the inflammatory stimulus, but the up-regulation of other inflammatory genes, e.g., cytokines and chemokines, such as interleukin-8, was not affected. On the other hand, HLA genes were increased by AZT. Notably, secreted IL-8 protein levels did not reflect mRNA levels, and were, in fact, higher after AZT pretreatment in cultures exposed to the inflammatory stimulus, suggesting that AZT can affect inflammatory pathways other than by altering gene expression. These findings suggest that the specific effects of AZT on inflamed and non-inflamed airway epithelia are likely relevant to its clinical activity, and their apparent complexity may help explain the diverse immunomodulatory roles of macrolides.
Ribeiro
Carla Maria P
CM
Cystic Fibrosis Center, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States of America. carla_ribeiro@med.unc.edu
Hurd
Harry
H
Wu
Yichao
Y
Martino
Mary E B
ME
Jones
Lisa
L
Brighton
Brian
B
Boucher
Richard C
RC
O'Neal
Wanda K
WK
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017963:Azithromycin / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects*; D004847:Epithelial Cells / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation; D016209:Interleukin-8 / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects*; D008055:Lipids / Q000737:chemistry*; D008168:Lung / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D055271:Mucin 5AC / Q000378:metabolism; D011552:Pseudomonas Infections / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism
D000900:Anti-Bacterial Agents; D016209:Interleukin-8; D008055:Lipids; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D017963:Azithromycin; D020780:Matrix Metalloproteinase 9