title: The periodontal pathogen Porphyromonas gingivalis sensitises human blood platelets to epinephrine.
abstract: Recent studies indicate connections between periodontitis and atherothrombosis, and the periodontal pathogen Porphyromonas gingivalis has been found within atherosclerotic lesions. P. gingivalis-derived proteases, designated gingipains activate human platelets, probably through a "thrombin-like" activity on protease-activated receptors (PARs). However, the potential interplay between P. gingivalis and other physiological platelet activators has not been investigated. The aim of this study was to elucidate consequences and mechanisms in the interaction between P. gingivalis and the stress hormone epinephrine. By measuring changes in light transmission through platelet suspensions, we found that P. gingivalis provoked aggregation, whereas epinephrine alone never had any effect. Intriguingly, pre-treatment of platelets with a low, sub-threshold number of P. gingivalis (i.e. a density that did not directly provoke platelet aggregation) resulted in a marked aggregation response when epinephrine was added. This synergistic action was not inhibited by the cyclooxygenas inhibitor aspirin. Furthermore, fura-2-measurements revealed that epinephrine caused an intracellular Ca(2+) mobilization in P. gingivalis pre-treated platelets, whereas epinephrine alone had no effect. Inhibition of the arg-specific gingipains, but not the lys-specific gingipains, abolished the aggregation and the Ca(2+) response provoked by epinephrine. Similar results were achieved by separate blockage of platelet alpha(2)-adrenergic receptors and PARs. In conclusion, the present study shows that a sub-threshold number of P. gingivalis sensitizes platelets to epinephrine. We suggest that P. gingivalis-derived arg-specific gingipains activates a small number of PARs on the surface of the platelets. This leads to an unexpected Ca(2+) mobilization and a marked aggregation response when epinephrine subsequently binds to the alpha(2)-adrenergic receptor. The present results are consistent with a direct connection between periodontitis and stress, and describe a novel mechanism that may contribute to pathological platelet activation.
authors.affiliation: Department of Clinical and Experimental Medicine, Division of Clinical Chemistry, Cardiovascular Inflammation Research Centre, Sweden. marny@ibk.liu.se
authors.forename: M
authors.initials: M
authors.lastname: Nylander
authors.forename: T L
authors.initials: TL
authors.lastname: Lindahl
authors.forename: T
authors.initials: T
authors.lastname: Bengtsson
authors.forename: M
authors.initials: M
authors.lastname: Grenegård
mesh_terms: D018829:Adhesins, Bacterial / Q000502:physiology*; D001081:Apyrase / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000382:microbiology*; D020013:Calcium Signaling / Q000187:drug effects; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000502:physiology*; D004837:Epinephrine / Q000494:pharmacology*; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D066298:In Vitro Techniques; D007976:Leupeptins / Q000494:pharmacology; D009391:Nephelometry and Turbidimetry; D009842:Oligopeptides / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000472:pathogenicity / Q000502:physiology*; D011480:Protease Inhibitors / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D011799:Quinazolines / Q000494:pharmacology; D044462:Receptors, Proteinase-Activated / Q000187:drug effects / Q000502:physiology; D014774:Virulence; D015016:Yohimbine / Q000494:pharmacology
chemical_list: D018829:Adhesins, Bacterial; D016861:Cyclooxygenase Inhibitors; D000080867:Gingipain Cysteine Endopeptidases; D007976:Leupeptins; C415424:N3-cyclopropyl-7-((4-(1-methylethyl)phenyl)methyl)-7H-pyrrolo(3, 2-f)quinazoline-1,3-diamine; D009842:Oligopeptides; D010446:Peptide Fragments; D010975:Platelet Aggregation Inhibitors; D011480:Protease Inhibitors; D011758:Pyrroles; D011799:Quinazolines; D044462:Receptors, Proteinase-Activated; C455901:alanyl-tyrosyl-prolyl-glycyl-lysyl-phenylalanine; C082835:thrombin receptor peptide (42-47); D015016:Yohimbine; D003546:Cysteine Endopeptidases; D001081:Apyrase; C032854:leupeptin; D001241:Aspirin; D004837:Epinephrine