Prevention of P2 Receptor-Dependent Thrombocyte Activation by Pore-Forming Bacterial Toxins Improves Outcome in a Murine Model of Urosepsis

• Main Authors: Mette G. Christensen, Nanna Johnsen, Marianne Skals, Aimi D. M. Hamilton, Peter Rubak, Anne-Mette Hvas, Helle Praetorius
• Format: Article
• Language: English
• Published: MDPI AG 2020-08-01
• Series: International Journal of Molecular Sciences
• Subjects: P2Y₁; P2Y₁₂; sepsis; <i>Escherichia</i> <i>coli</i>; HlyA; thrombocytes
• Online Access: https://www.mdpi.com/1422-0067/21/16/5652

Urosepsis is a potentially life-threatening, systemic reaction to uropathogenic bacteria entering the bloodstream of the host. One of the hallmarks of sepsis is early thrombocyte activation with a following fall in circulating thrombocytes as a result of intravascular aggregation and sequestering of thrombocytes in the major organs. Development of a thrombocytopenic state is associated with a poorer outcome of sepsis. Uropathogenic <i>Escherichia coli</i> frequently produce the pore-forming, virulence factor α-haemolysin (HlyA), of which the biological effects are mediated by ATP release and subsequent activation of P2 receptors. Thus, we speculated that inhibition of thrombocyte P2Y₁ and P2Y₁₂ receptors might ameliorate the septic response to HlyA-producing <i>E. coli</i>. The study combined in vitro measurements of toxin-induced thrombocyte activation assessed as increased membrane abundance of P-selectin, fibronectin and CD63 and data from in vivo murine model of sepsis-induced by HlyA-producing <i>E. coli</i> under infusion of P2Y₁ and P2Y₁₂ antagonists. Our data show that the P2Y₁ receptor antagonist almost abolishes thrombocyte activation by pore-forming bacterial toxins. Inhibition of P2Y₁, by constant infusion of MRS2500, markedly increased the survival in mice with induced sepsis. Moreover, MRS2500 partially prevented the sepsis-induced depletion of circulating thrombocytes and dampened the sepsis-associated increase in proinflammatory cytokines. In contrast, P2Y₁₂ receptor inhibition had only a marginal effect in vivo and in vitro. Taken together, inhibition of the P2Y₁ receptor gives a subtle dampening of the thrombocyte activation and the cytokine response to bacteraemia, which may explain the improved survival observed by P2Y₁ receptor antagonists.