Accumulation of extracellular ATP protects against acute reperfusion injury in rat heart endothelial cells

Dursun Gündüz, Justus-Liebig-Universität Giessen, German
Sascha A. Kasseckert, Justus-Liebig-Universität Giessen, German
Frauke V. Härtel, Justus-Liebig-Universität Giessen, German
Muhammad Aslam, Justus-Liebig-Universität Giessen, German
Yaser Abdallah, Aga Khan University
Matthias Schäfer, Justus-Liebig-Universität Giessen, German
Hans Michael Piper, Justus-Liebig-Universität Giessen, German
Thomas Noll, Justus-Liebig-Universität Giessen, German
Claudia Schäfer, Justus-Liebig-Universität Giessen, German

This work was published before the author joined Aga Khan University.



Ischemia–reperfusion provokes barrier failure of the coronary microvasculature, leading to myocardial edema development that jeopardizes functional recovery of the heart during reperfusion. Here, we tested whether adenosine 5′-triphosphate (ATP), either exogenously applied or spontaneously released during reperfusion, protects the endothelial barrier against an imminent reperfusion injury and whether interventions preventing ATP breakdown augment this protective ATP effect. Methods: Cultured microvascular coronary endothelial monolayers and isolated-perfused hearts of rat were used.

Results: After ischemic conditions were induced, reperfusion of endothelial monolayers activated the endothelial contractile machinery and caused intercellular gap formation. It also led to the release of ATP. When its breakdown was inhibited by 6-N,N-diethyl-β,γdibromomethylene-D-ATP (ARL 67156; 100 μM), a selective ectonucleotidase inhibitor, contractile activation and gap formation were significantly reduced. Reperfusion in the presence of exogenously added ATP (10 μM) plus ARL caused an additional reduction of both aforementioned effects. In contrast, elevation of ATP degradation by apyrase (1 U/ml), a soluble ectonucleotidase, or addition of adenosine (10 μM) provoked an increase in gap formation during reperfusion that could be completely inhibited by 8-phenyltheophylline (8-PT; 10 μM), an adenosine receptor antagonist. In Langendorff-perfused rat hearts, the reperfusion-induced increase in water content was significantly reduced by ARL plusATP. Under conditions favouring ATP degradation, an increase in myocardial edema was observed that could be blocked by 8-PT.

Conclusion: ATP, either released from cells or exogenously applied, protects against reperfusion-induced failure of the coronary endothelial barrier. Inhibition of ATP degradation enhances the stabilizing effect of ATP on barrier function.