Arachidonic acid induces DNA-fragmentation in human polymorphonuclear neutrophil granulocytes.
We analyzed DNA-fragmentation in human polymorphonuclear neutrophil granulocytes (PMNs) from healthy donors after addition of exogenous arachidonic acid (AA) and eicosapentaenoic acid (EPA) by flow cytometry (propidium iodide staining of DNA and DNA strand break detection). The PMNs were incubated from 30 min up to 48 hours in RPMI 1640 which was supplemented with different concentrations of AA or EPA (5-40 microM). In contrast to EPA the addition of AA led to a significant increase in apoptosis up to 67.8% compared to the RPMI-control whereas the addition of EPA led to an inhibition of DNA-fragmentation. When the cells were incubated with MK 886 (1 microM, inhibitor of leukotriene biosynthesis) an increase in DNA-fragmentation (up to 63.3%) was observed. Conversely, in the presence of indomethacin (1 microM, inhibitor of prostanoid synthesis) an inhibition in DNA-fragmentation (up to 60.9%) occurred. Furthermore, preincubation of PMNs with pentoxifylline (1mM, phosphodiesterase inhibitor) reduced AA-stimulated DNA-fragmentation up to 43.4%. These data provide evidence for the involvement of AA and distinct AA metabolites in the regulation of apoptosis in human PMNs.
Köller
M
M
Department of Surgery, BG Klinik Bergmannsheil, Bochum, Germany.
Wachtler
P
P
Dávid
A
A
Muhr
G
G
König
W
W
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology*; D004396:Coloring Agents; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects* / Q000502:physiology; D015118:Eicosapentaenoic Acid / Q000378:metabolism / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D007249:Inflammation / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D015289:Leukotrienes / Q000096:biosynthesis; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D010431:Pentoxifylline / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011419:Propidium; D002722:Chlorobenzenes
D016718:Arachidonic Acid; D004396:Coloring Agents; D016861:Cyclooxygenase Inhibitors; D015118:Eicosapentaenoic Acid; D007211:Indoles; D007213:Indomethacin; D015289:Leukotrienes; D016859:Lipoxygenase Inhibitors; D010431:Pentoxifylline; D010726:Phosphodiesterase Inhibitors; D011419:Propidium; C060893:MK-886; D002722:Chlorobenzenes