Macrolides are known for immunomodulatory and anti-inflammatory actions. To analyze Clarithromycin (CAM) mechanism of action, we examined the effects of CAM on translatome of LPS-treated or untreated B-lymphoblastoid cells (BJAB). Translatome analyses were performed in 4 groups, i.e., LPS-treated or untreated (saline) BJAB, and cells further added with saline or CAM:LPS-saline(S), LPS-CAM, S-CAM, S-S. The cytoplasmic extracts were fractionated by sucrose gradient-centrifugation and divided into ten fractions, for RNA extraction. The RNAs extracted from polysome fractions were subjected to DNA microarray analysis.
GenMAPP Pathway-analysis (Filgen) showed significant gene expression changes in LPS-S, LPS-CAM and S-CAM. Analysis of mRNA-processing pathway showed that many gene expressions were modulated by CAM: 44 kinds of genes (factors associated with transcription factors, mRNA transcription and splicing factors, SR-proteins, and maturation and stability factors). Significant expression changes were seen in inflammation-related genes, such as IL-3, IL-5, EGF-EGFR, and EPO receptor pathway, in LPS-CAM. As mentioned above, it is likely that CAM controls inflammation pathway specifically in LPS-pretreated cells.
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