Mono(2-ethylhexyl) phthalate induces both pro- and anti-inflammatory responses in rat alveolar macrophages through crosstalk between p38, the lipoxygenase pathway and PPARalpha.
Airway inflammation is important in asthma pathogenesis. Recent epidemiological data have indicated an association between asthma symptoms in children and exposure to di(2-ethylhexyl) phthalate (DEHP). Thus, we have studied inflammatory responses in primary rat alveolar macrophages (AMs) after exposure to mono(2-ethylhexyl) phthalate (MEHP), the major primary metabolite of DEHP. First, we show that MEHP induces a dose-dependent release of the pro-inflammatory tumour necrosis factor-alpha (TNF-alpha) in AMs, giving a maximal (5-fold) increase at 0.7 mM. This concentration also induced some cell death. MEHP also induced phosphorylation of MAPK p38, while the p38 inhibitor SB 202190 reduced MEHP-induced TNF-alpha, suggesting a p38-dependent cytokine production. Next, we elucidated possible effects of MEHP on the 5-lipoxygenase (5-LO) pathway and found that MEHP caused increased leukotriene (LTB(4)) release. Further, we found that the 5-LO inhibitor nordihydrogualaretic acid (NDGA) significantly reduced both MEHP-induced TNF-alpha release and MEHP-induced formation of reactive oxygen species (ROS), supporting an involvement of the 5-LO pathway in MEHP induced inflammatory reactions. Last, we found that MK-886, a known inhibitor of peroxisome proliferator-activated receptor alpha (PPARalpha), increased the MEHP-induced TNF-alpha response. This indicates that MEPH-PPARalpha binding mediates an anti-inflammatory signal.
Rakkestad
Kirsten E
KE
Department of Air Pollution and Noise, Division of Environmental Medicine, Norwegian Institute of Public Health, Oslo, Norway.
Holme
Jørn A
JA
Paulsen
Ragnhild E
RE
Schwarze
Per E
PE
Becher
Rune
R
D000818:Animals; D000893:Anti-Inflammatory Agents; D016923:Cell Death / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects; D002469:Cell Separation; D051546:Cyclooxygenase 2 / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D007249:Inflammation / Q000139:chemically induced* / Q000473:pathology; D015289:Leukotrienes / Q000096:biosynthesis; D008084:Lipoxygenase / Q000502:physiology*; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000473:pathology*; D008297:Male; D047493:PPAR alpha / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D011921:Rats, Inbred WKY; D017208:Rats, Wistar; D017382:Reactive Oxygen Species; D020239:Receptor Cross-Talk / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000502:physiology*
D000893:Anti-Inflammatory Agents; D016207:Cytokines; D015289:Leukotrienes; D047493:PPAR alpha; D017382:Reactive Oxygen Species; D014409:Tumor Necrosis Factor-alpha; D004051:Diethylhexyl Phthalate; D008084:Lipoxygenase; D051546:Cyclooxygenase 2; D048051:p38 Mitogen-Activated Protein Kinases