Effects of montelukast on M2-related cytokine and chemokine in M2 macrophages.
OBJECTIVE
Asthma is a chronic airway inflammatory disease mediated by T-helper (Th)2 cells. Montelukast (trade name Singulair) is a cysteinyl leukotriene receptor antagonist used for asthma treatment. Mirroring Th1-Th2 polarization, two distinct states of macrophages have been recognized: the classically activated (M1) macrophages and the alternatively activated (M2) macrophages. M2 polarization is known to be a response to the Th2 cytokines; however, the effects of montelukast on M2 macrophages have not been well characterized. The aim of the present study was to investigate the effects of montelukast on the expression of cytokines and chemokines in M2-like macrophages, and to explore possible intracellular signaling pathways.


METHODS
The human monocytic leukemia cell line THP-1 and human monocytes from healthy donors were cultured with interleukin-4 for M2 polarization, and then the cells were pretreated with or without montelukast before lipopolysaccharide (LPS) stimulation. Supernatants were collected to determine interleukin-10, I-309/CCL1, and MDC/CCL22 levels by enzyme-linked immunosorbent assay. Intracellular signaling was investigated using nuclear factor (NF)-κB inhibitors, mitogen-activated protein kinase (MAPK) inhibitors, and western blot analysis.


RESULTS
LPS-induced interleukin-10 and I-309/CCL1 expression was significantly suppressed by montelukast in THP-1-derived and human monocyte-derived M2 macrophages after LPS stimulation. MDC/CCL22 expression was only significantly suppressed by montelukast in THP-1-derived M2 macrophages after 48 hours of incubation. In western blot analysis, montelukast was able to suppress LPS-induced MAPK-phospho-p38 and NF-κB-phospho-p65 expression.


CONCLUSIONS
Montelukast suppressed LPS-induced M2-related cytokines and chemokines in alternatively activated macrophages, and the effects might be mediated through the MAPK-p38 and NF-κB-p65 pathways.
Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Laboratory Medicine, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Yi-Ching
YC
Lin
Department of Radiation Oncology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Ming-Yii
MY
Huang
Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Pediatrics, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung, Taiwan.
Min-Sheng
MS
Lee
Division of Cardiac Surgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.
Chong-Chao
CC
Hsieh
Division of Cardiology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.
Hsuan-Fu
HF
Kuo
Department of Pediatrics, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung, Taiwan; Ta-Kuo Clinic, Kaohsiung, Taiwan. Electronic address: kuochanghung.dr@gmail.com.
Chang-Hung
CH
Kuo
Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Pediatrics, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung, Taiwan; Department of Pediatrics, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Kaohsiung Medical University Aim for Top Universities Grant, Taiwan. Electronic address: pedhung@gmail.com.
Chih-Hsing
CH
Hung
D000085:Acetates / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D054402:Chemokine CCL1 / Q000378:metabolism; D054422:Chemokine CCL22 / Q000378:metabolism; D018925:Chemokines / Q000378:metabolism*; D003521:Cyclopropanes; D016207:Cytokines / Q000378:metabolism*; D006801:Humans; D016753:Interleukin-10 / Q000378:metabolism; D015847:Interleukin-4; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D009000:Monocytes / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism; D011804:Quinolines / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D013440:Sulfides; D000074084:THP-1 Cells / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
D000085:Acetates; C065971:CCL1 protein, human; C515482:CCL22 protein, human; D054402:Chemokine CCL1; D054422:Chemokine CCL22; D018925:Chemokines; D003521:Cyclopropanes; D016207:Cytokines; C508609:IL10 protein, human; D008070:Lipopolysaccharides; D016328:NF-kappa B; D011804:Quinolines; D013440:Sulfides; D016753:Interleukin-10; D015847:Interleukin-4; D048051:p38 Mitogen-Activated Protein Kinases; C093875:montelukast
Chemokine; M2 macrophage; Mitogen-activated protein kinase; Montelukast; Nuclear factor-κB