PLA(2) stimulation of Na(+)/H(+) antiport and proliferation in rat aortic smooth muscle cells.
The proliferative properties and the ability to stimulate the Na(+)/H(+) antiport activity of a secretory phospholipase A(2) were studied in rat aortic smooth muscle cells in culture. The requirement of the enzymatic activity of phospholipase A(2) to elicit mitogenesis was assessed by the use of ammodytin L, a Ser(49) phospholipase A(2) from the venom of Vipera ammodytes, devoid of hydrolytic activity. We propose that the proliferative effect is mediated by the same transduction pathway for both proteins. In particular, 1) both secretory phospholipase A(2) and ammodytin L stimulated thymidine incorporation in a dose-dependent manner; 2) both proteins affected the cell cycle, as assessed by cell growth and fluorescence-activated cell sorting experiments; 3) both phospholipase A(2) and ammodytin L increased intracellular pH, a permissive factor for cell proliferation, through activation of the Na(+)/H(+) antiport; 4) ammodytin L was able to displace the (125)I-labeled phospholipase A(2) from specific binding sites in a concentration range consistent with that capable of eliciting a cellular response; and 5) the inhibition by heparin was similar for both proteins, taking into account the ratio of heparin to protein. In conclusion, the enzymatic activity of phospholipase A(2) is not required for the stimulation of mitogenesis. The inhibitory effect of heparin combined with its therapeutic potential could help to clarify the role of phospholipase A(2) in the pathogenesis of several preinflammatory situations.
Rufini
S
S
Department of Biology, University of Rome "Tor Vergata," 00133 Rome, Italy. rufini@uniroma2.it
de Vito
P
P
Balestro
N
N
Pescatori
M
M
Luly
P
P
Incerpi
S
S
D000818:Animals; D001011:Aorta / Q000166:cytology* / Q000378:metabolism*; D001667:Binding, Competitive; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism; D014757:Viper Venoms / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology
D004791:Enzyme Inhibitors; D017923:Sodium-Hydrogen Exchangers; D014757:Viper Venoms; C065419:ammodytin L; D006493:Heparin; D010741:Phospholipases A; D013936:Thymidine