Involvement of mitogen-activated protein kinase and NF-kappaB activation in Ca2+-induced IL-8 production in human mast cells.
Interleukin-8 (IL-8) is a potent proinflammatory chemokine that plays an important role in inflammation by activating and recruiting neutrophils, lymphocytes, and eosinophils. To demonstrate the effect of intracellular Ca(2+) on IL-8 production and related signaling, we stimulated human mast cell line HMC-1 with either calcium ionophore A23187 or thapsigargin. Increase of intracellular Ca(2+) resulted in inducing IL-8 gene expression and protein secretion, and addition of EGTA or BAPTA/AM before Ca(2+) stimulation inhibited the induction of IL-8 production. Intracellular Ca(2+) triggered the activation of mitogen-activated protein kinase (MAPK) in HMC-1, especially p42 and p44 isoforms of extracellular signal-regulated kinase (ERK) and p38 MAPK, but not c-Jun N-terminal kinase (JNK). Pretreatment of MAPK inhibitors (PD98059 and SB203580) markedly blocked Ca(2+)-induced IL-8 production from cells, and anti-inflammatory drugs, such as dexamethasone and cyclosporin A, partially inhibited the activation of ERK1/2. We determined that increased Ca(2+) activates the nuclear translocation of the transcription factor NF-kappaB. NF-kappaB inhibitors blocked the ability of Ca(2+) to induce IL-8 production, and the activation of NF-kappaB was required for intracellular Ca(2+)-induced up-regulation of IL-8. These results suggest that increased intracellular Ca(2+) stimulated p38 and ERK1/2 MAPK signaling cascades result in NF-kappaB activation and IL-8 production in HMC-1 cells. This study is the first to identify the intracellular signaling pathways involved in the Ca(2+)-mediated up-regulation of IL-8 synthesis and release from HMC-1 cells.
Kim
Mi Sun
MS
Department of Pharmacology, College of Oriental Medicine, Kyung Hee University, 1 Hoegi-Dong, Dongdaemun-Gu, Seoul 130-701, Republic of Korea.
Lim
Woon Ki
WK
Park
Rae Kil
RK
Shin
Taekyun
T
Yoo
Young-Hyun
YH
Hong
Seung-Heon
SH
An
Nyeon-Hyoung
NH
Kim
Hyung Min
HM
D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics; D008407:Mast Cells / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D048308:Mitogen-Activated Protein Kinase 14 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism*; D019284:Thapsigargin / Q000494:pharmacology; D015854:Up-Regulation
D004791:Enzyme Inhibitors; D016209:Interleukin-8; D016328:NF-kappa B; D000001:Calcimycin; D019284:Thapsigargin; D048049:Extracellular Signal-Regulated MAP Kinases; D048308:Mitogen-Activated Protein Kinase 14; D002118:Calcium