title: Embryonal cardiotoxicity of the Helicobacter pylori lipopolysaccharide.
abstract: Helicobacter (H.) pylori is the causative agent of the peptic ulcer disease and a co-factor in the development of gastric malignancies. Recently, it has been maintained that chronic H. pylori infections in adults are linked to a higher risk of coronary heart diseases. In this respect, the acute toxic effects of the H. pylori lipopolysaccharide (LPS) on embryonal cardiomyocytes at different developmental stages was evaluated. White Leghorn chick embryos and smooth (S)--form NCTC 11637 strain H. pylori organisms were used. Both whole heath-killed H. pylori suspensions (3.10(6) bacteria/egg) and isolated S-LPS (500 ng/egg) or S-Lipid A (500 ng/egg) were non-lethal to 4-day embryos, becoming moderately lethal (5% to 30%) to 6- and 8-day embryos and highly lethal (> 90%) to 10- to 17-day embryos. The contractile activity of isolated atrial fragments from 10-day embryos was completely inhibited, within 5 min, following treatments with heath-killed H. pylori (3 x 10(6)/ml), or S-LPS (500 ng/ml), or S-Lipid A (500 ng/ml); the block determined by S-LPS and S-Lipid A was irreversible, while the block by bacterial suspensions was completely reversible upon withdrawal. Following a 24-hour treatment with S-LPS or S-Lipid A of single-cell cultures of cardiomyocytes (isolated from 10-day embryos) a dose-dependent cell loss was observed, as assessed by total protein dosage and direct counting of adherent cells. Propidium Iodide/Annexin V FACS-analysis confirmed the occurrence of cellular necrosis, but did not show any evidence of apoptotic processes. The release of superoxide anion radicals by cultured cardiomyocytes was as follows: S-Lipid A (25 micrograms/ml) > S-LPS (25 micrograms/ml) > heath killed H. pylori suspensions (3 x 10(6)/ml); control cultures did not release detectable amounts of superoxide anion radicals. Furthermore, cultured cardiomyocytes produced increased amounts of NO (N-monomethylarginine-inhibitable) following stimulation with S-LPS (25 micrograms/ml) or S-Lipid A (25 micrograms/ml) (but not heath killed H. pylori 3 x 10(6)/ml suspensions). Under all the above experimental conditions S-polysaccharide proved to be non-toxic. Concluding, H. pylori LPS is relatively non-toxic to the less differentiated cardiomyocytes; cardiomyocytes which are more advanced in their biochemical differentiation become highly sensitive to LPS and produce ROS and NO. ROS are probably responsible for the early toxic actions, while both ROS and NO are likely to be involved in the later degenerative/necrotic effects.
authors.affiliation: Department of Human Anatomy and Histology, University of Bari, Policlinico, piazza, Giulio, Cesare, 1-70124, Bari, Italy. ma.panaro@anatomia.uniba.it
authors.forename: M A
authors.initials: MA
authors.lastname: Panaro
authors.forename: L
authors.initials: L
authors.lastname: Amati
authors.forename: M
authors.initials: M
authors.lastname: Sisto
authors.forename: L
authors.initials: L
authors.lastname: Caradonna
authors.forename: S
authors.initials: S
authors.lastname: Lisi
authors.forename: V
authors.initials: V
authors.lastname: Mitolo
authors.forename: D
authors.initials: D
authors.lastname: Fumarola
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002642:Chick Embryo; D006321:Heart / Q000187:drug effects / Q000196:embryology*; D006331:Heart Diseases / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D016480:Helicobacter pylori / Q000737:chemistry* / Q000235:genetics; D008070:Lipopolysaccharides / Q000633:toxicity*; D009206:Myocardium / Q000473:pathology; D009569:Nitric Oxide / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
chemical_list: D008070:Lipopolysaccharides; D013481:Superoxides; D009569:Nitric Oxide