Pseudomonas aeruginosa induces apoptosis in human endothelial cells.
Pseudomonas aeruginosa has been shown to enter into human endothelial cells in vitro. To ascertain the effects of bacterial intracellular (IC) infection, endothelial cells were exposed to PAK and PAO-1 strains for 1 h and treated with gentamicin in culture medium for different periods. P. aeruginosa induced a significant production of superoxide and hydrogen peroxide by endothelial cells. Concentrations of IC bacteria were reduced progressively with time and no viable PAO-1 was detected at 24 h after infection. However, IC infection led to killing of 32.2%+/-2.9 and 51.8%+/-3.5 of the cells infected with PAK and PAO-1, respectively, as determined by the MTT assay. By three criteria (transmission electron microscopy, DNA electrophoresis and reactivity with annexin V) infected cells exhibited features of apoptosis. Treatment of infected cells with anti-oxidants (catalase, tocopherol and N -acetyl-L-cysteine) significantly decreased the percentage of cell death. In contrast, treatment with aminoguanidine, an inhibitor of inducible NO synthase, increased significantly the killing of PAO-1 infected cells. Based on these results we speculate that in response to P. aeruginosa infection, endothelial cells increase the production of reactive oxygen intermediates to eliminate IC pathogens, but cells do not resist the oxidative stress and die by apoptosis.
Valente
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Department of Microbiology and Immunology, State University of Rio de Janeiro, Rio de Janeiro, Brazil.
Assis
M C
MC
Alvim
I M
IM
Pereira
G M
GM
Plotkowski
M C
MC
D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000502:physiology*; D002374:Catalase / Q000494:pharmacology; D002460:Cell Line; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D004727:Endothelium / Q000166:cytology / Q000382:microbiology*; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D005562:Formazans / Q000737:chemistry; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D008070:Lipopolysaccharides; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010617:Phenanthridines / Q000737:chemistry; D011552:Pseudomonas Infections / Q000473:pathology*; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000235:genetics / Q000472:pathogenicity*; D017382:Reactive Oxygen Species / Q000502:physiology; D013778:Tetrazolium Salts / Q000737:chemistry; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D005562:Formazans; D006146:Guanidines; D008070:Lipopolysaccharides; D010617:Phenanthridines; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; D014810:Vitamin E; C079782:MTT formazan; C058428:hydroethidine; D002374:Catalase; C004479:pimagedine; D000111:Acetylcysteine