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Cytosolic phospholipase A2 and activity upon stimulation with A23187, trypsin and TNF-2 (cPLA2) and calcium-independent2 (iPLA2) in the human mast cell line HMC-1
University of Kalmar, School of Pure and Applied Natural Sciences. (Christina Gustafson-Svärd)
Klinisk och experimentell kirurgi, avd för kirurgi Hälsouniversitetet Linköping. (Johan D Söderholm)
University of Kalmar, School of Pure and Applied Natural Sciences. (Christina Gustafson-Svärd)
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Background

 

 
: Mast cells (MC) are considered to be involved in the pathogenesis of Crohn´s disease (CD). A key regulatory event in the production of eicosanoids in MC is the mobilization of arachidonic acid (AA) by the action of phospholipases A2 (PLA2s). The expression of protease activated receptor-2 (PAR-2) is increased on intestinal mast cells in CD, but it is not known if this receptor may influence the PLA2 activity in MC. Although extensively studied in rodent mast cells, not much is known about the PLA2s in human mast cells. The principle aims of the present study were to investigate if the cell line HMC-1 could serve as a model for studies on cytosolic PLA2 (cPLA2) and calcium-independent PLA2 (iPLA2) in human mast cells, and if stimulation of PAR-2 may influence the activity of the two enzymes. Methods: A human mast cell line, HMC-1, was used for the studies. PLA2 activity was analyzed by quantification of radiolabeled fatty acids. PLA2 expression was investigated by RT-PCR and immunocytochemistry. Results: A23187 and the PAR-2-activator trypsin induced a fatty acid release that was not restricted to AA. Only the A23187-induced AArelease was reduced by a combined cPLA2 and iPLA2 inhibitor and by a specific iPLA2 inhibitor. The protein expression of cPLA2, as determined by immunocytochemistry, was very low compared to the expression of iPLA2. TNF-α specifically increased the cPLA2 mRNA expression but had no effect on the protein expression of the PLA2s, as determined by immunocytochemistry. Conclusions: Stimulation with a calcium ionophore may activate iPLA2-dependent AA release, but neither cPLA2 nor iPLA2 seems to be involved in PAR-2-stimulated AA release. TNF-α stimulates cPLA2 mRNA expression, but does not influence the immunocytochemically detected protein level. The expression of cPLA2 and iPLA2 in HMC-1 cells, together with its ability to release radiolabeled fatty acids upon stimulation, makes this cell line a promising model for further studies on cPLA2 and iPLA2 in human MCs. 

National Category
Natural Sciences
Research subject
Natural Science, Biomedical Sciences
Identifiers
URN: urn:nbn:se:lnu:diva-6509OAI: oai:DiVA.org:lnu-6509DiVA, id: diva2:326844
Available from: 2010-06-24 Created: 2010-06-24 Last updated: 2010-10-14Bibliographically approved

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Christerson, Ulrika

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