Melatonin inhibits lipopolysaccharide‐induced CC chemokine subfamily gene expression in human peripheral blood mononuclear cells in a microarray analysis

Abstract

Melatonin possesses a number of important biologic activities including oncostatic, anti-oxidant, and immunostimulatory actions. This study was designed to assess the effects of melatonin on inflammation-related gene expression in lipopolysaccharide (LPS)-stimulated human peripheral blood mononuclear cells (PBMCs), using CombiMatrix 2K Human Inflammation chip. After pretreatment with melatonin (100 microm) for 4 hr, cells were incubated with LPS (1 microg/mL) for 24 hr. We compared gene expression profiles between LPS-treated, melatonin-treated, LSP/melatonin-treated, and control groups. LPS induced the upregulation of 95 genes, compared with controls. Melatonin pretreatment in LPS-stimulated PBMCs suppressed the expression of 23 genes more than twofold. Interestingly, melatonin showed a suppressive effect on the expression of CC chemokine subfamily genes, including CCL2/MCP1, CCL3/MIP1 alpha, CCL4/MIP1 beta, CCL5/RANTES, CCL8/MCP2, CCL20/MDC, and CCL22/MIP3 alpha, in LPS-stimulated PBMCs. This result was confirmed by reverse transcriptase polymerase chain reaction. Among the CC chemokine subfamily genes, particularly, the expression of CCL2 and CCL5 was markedly downregulated by melatonin in LPS-stimulated PBMCs. The secretion levels of CCL2 and CCL5 were measured using enzyme-linked immunosorbent assay. Stimulation of PBMCs by LPS induced the secretion of CCL2 (2334.3 +/- 161.4 pg/mL, mean +/- S.E.M.), whereas melatonin pretreatment (153.0 +/- 3.8 pg/mL) inhibited the LPS-induced secretion of CCL2. Melatonin pretreatment (2696.2 +/- 385.3 pg/mL) also inhibited the LPS-induced secretion of CCL5 (4679.6 +/- 107.5 pg/mL). Taken together, these results suggest that melatonin may have a suppressive effect on LPS-induced expression of CC chemokine genes, especially CCL2 and CCL5, which may explain its beneficial effects in the treatment of various inflammatory conditions.

Transcriptome analysis of porcine PBMCs after in vitro stimulation by LPS or PMA/ionomycin using an expression array targeting the pig immune response

“…Up-regulation of IL8 has already been reported in pig PBMCs [39,40] and amnion [41] after bacterial infection. In human, stimulation of PBMCs with LPS induces the secretion of CCL2 [42], CXCL3 and CXCL2 [43]. CXCL5 is up-regulated in LPS-challenged bovine mammary epithelial cells [44].…”

Microarray Analysis of Gene Expression Profiles in Response to Treatment with Melatonin in Lipopolysaccharide Activated RAW 264.7 Cells

“…In a previous study, we demonstrated that melatonin inhibited LPS-induced CC chemokine subfamily gene expression in human peripheral blood mononuclear cells using a CombiMatrix 2k Human Inflammation chip [12]. In the present study, we investigated the effect of melatonin on global gene expression in LPS-stimulated RAW 264.7 cells, using a 24K cDNA microarray.…”

A nested case–control study of the association between melatonin secretion and incident myocardial infarction

“…Third, melatonin has an important role in regulating the immune system and therefore inflammation. Via the melatonin receptor 1A (MTNR1A) melatonin reduces the release of interleukin-8 and Tumor necrosis factor (TNF-α) from neutrophils and also markedly reduces the production of CC chemokines by monocytes 21. Melatonin also inhibits activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) in endothelial cells and reduces leucocyte adhesion to the endothelium, presumably through inhibited expression of Intercellular Adhesion Molecule 1 (ICAM-1) and P-selectin on endothelial cells 22 23.…”