Lopap: a non-inflammatory and cytoprotective molecule in neutrophils and endothelial cells.
Lopap (Lonomia obliqua prothrombin activator protease) is a member of the lipocalin family isolated from the extract of L obliqua bristles. Lopap displays serine protease-like activities, including coagulation disturbance, cytokine secretion and antiapoptotic activity in human cultured endothelial cells. Here, we have investigated the effects of the recombinant protein (rLopap) on the inflammatory and apoptotic processes of neutrophils and endothelial cells from male Wistar rats. We found that rLopap did not induce in vivo leukocyte-endothelial interactions in the microvasculature, initial steps of leukocyte recruitment during inflammation. Incubation of rLopap with neutrophils or endothelial cells prevented apoptosis evoked by serum deprivation and induced nitric oxide (NO) production in both cell types, and increased the expression of ICAM-1 by endothelial cells. Simultaneous incubation of endothelial cells or neutrophils with rLopap and N omega-nitro-L-arginine methyl ester (L-NAME), a non-specific inhibitor of NO synthases, inhibited NO production and impaired the protection on apoptosis. Differently, incubation of endothelial cells with monoclonal antibody anti ICAM-1 did not change the protection on apoptosis evoked by rLopap. Together, these results indicate that rLopap does not display inflammatory properties in vivo but inhibits apoptosis of neutrophils and endothelial cells depending, at least in part, on NO production.
Waismam
Kaline
K
Department of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of São Paulo. Av. Prof Lineu Prestes 580, Bloco 13 B. São Paulo, SP, 05503-900, Brazil.
Chudzinski-Tavassi
Ana Marisa
AM
Carrijo-Carvalho
Linda C
LC
Pacheco
Mario T Fernandes
MT
Farsky
Sandra H P
SH
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002450:Cell Communication / Q000187:drug effects; D019610:Cytoprotection; D042783:Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D007249:Inflammation / Q000139:chemically induced; D015850:Interleukin-6 / Q000096:biosynthesis; D008297:Male; D009504:Neutrophils / Q000187:drug effects* / Q000502:physiology; D009569:Nitric Oxide / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D015850:Interleukin-6; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; C119835:Lonomia obliqua prothrombin activator protease; D012697:Serine Endopeptidases