Erythrocytes do not activate purified and platelet soluble guanylate cyclases even in conditions favourable for NO synthesis

Cell Communication and Signaling

Research

Cora Reiss¹  Ulrich Walter²  Iraida Sharina³  Emil Martin³  Stepan Gambaryan⁴  Hariharan Subramanian⁵  Andreas Friebe⁶  Linda Kehrer⁶  Igor Mindukshev⁷  Julia Sudnitsyna⁷  Natalia Rukoyatkina⁷ 

[1]Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany

[2]Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany

[3]German Centre for Cardiovascular Research (DZHK) RheinMain, Mainz, Germany

[4]Department of Internal Medicine, Division of Cardiology, University of Texas Houston Medical School, Houston, USA

[5]Institute of Clinical Biochemistry and Pathobiochemistry, University of Wuerzburg, Grombuehlstraße 12, D-97080, Wuerzburg, Germany

[6]Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Thorez pr. 44, St, 194223, Petersburg, Russia

[7]Center for Thrombosis and Hemostasis (CTH), University Medical Center Mainz, Mainz, Germany

[8]Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

[9]Institute of Physiology, University of Wuerzburg, Wuerzburg, Germany

[10]Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Thorez pr. 44, St, 194223, Petersburg, Russia

关键词: Erythrocytes;    Nitric oxide;    Soluble guanylate cyclase;    Platelets;    Hemoglobin;   

DOI  :  10.1186/s12964-016-0139-9

 received in 2016-06-05, accepted in 2016-08-01,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundDirect interaction between Red blood cells (RBCs) and platelets is known for a long time. The bleeding time is prolonged in anemic patients independent of their platelet count and could be corrected by transfusion of RBCs, which indicates that RBCs play an important role in hemostasis and platelet activation. However, in the last few years, opposing mechanisms of platelet inhibition by RBCs derived nitric oxide (NO) were proposed. The aim of our study was to identify whether RBCs could produce NO and activate soluble guanylate cyclase (sGC) in platelets.MethodsTo test whether RBCs could activate sGC under different conditions (whole blood, under hypoxia, or even loaded with NO), we used our well-established and highly sensitive models of NO-dependent sGC activation in platelets and activation of purified sGC. The activation of sGC was monitored by detecting the phosphorylation of Vasodilator Stimulated Phosphoprotein (VASPS239) by flow cytometry and Western blot. ANOVA followed by Bonferroni’s test and Student’s t-test were used as appropriate.ResultsWe show that in the whole blood, RBCs prevent NO-mediated inhibition of ADP and TRAP6-induced platelet activation. Likewise, coincubation of RBCs with platelets results in strong inhibition of NO-induced sGC activation. Under hypoxic conditions, incubation of RBCs with NO donor leads to Hb-NO formation which inhibits sGC activation in platelets. Similarly, RBCs inhibit activation of purified sGC, even under conditions optimal for RBC-mediated generation of NO from nitrite.ConclusionsAll our experiments demonstrate that RBCs act as strong NO scavengers and prevent NO-mediated inhibition of activated platelets. In all tested conditions, RBCs were not able to activate platelet or purified sGC.

【 授权许可】

CC BY   
© The Author(s). 2016