Abstract Number: PB0998
Meeting: ISTH 2021 Congress
Theme: Platelets and Megakaryocytes » Platelet Function and Interactions
Background: Cardiovascular diseases are the leading cause of death in the world. Platelets play a major role in cardiovascular events, binding to the damaged endothelium, activating and forming thrombi.
Aims: Evaluate the antiplatelet effect of hydroquinone derivatives targeting to mitochondria, in human platelets.
Methods: We carry out cytotoxicity studies and platelet aggregation screening to a series of compounds. The compound with the highest activity was compound JP-I (Figure 1). To evaluate the mechanism of action of compound JP-I on platelets, we performed techniques of: Flow cytometry for markers of platelet activation, ROS, mitochondrial membrane potential and intracellular calcium levels; platelet spreading by fluorescence microscopy; Intracellular ATP and ATP secretion by luminescence; and NADH in real time by Fluorimetry; AMPK-ACC signaling western blot, in vitro clot retraction and occlusion time with PFA-200 System. Concentrations close to the IC50 obtained in platelet aggregation were used.
Acknowledgment: This research was funded by ANID/CONICYT, FONDECYT grant N◦ 1180427 (EF), and 1180069 (RAM)Figure 1. JP-I compound chemical structure
Results: Compound JP-I does not present cytotoxicity and inhibits platelet aggregation, with IC50 values: 3.99 ± 1.43 μM (Thrombin Receptor Activator Peptide-6) and 7.16 ± 0.9 μM (Phorbol 12-myristate13-acetate). Decreases the expression of P-selectin, CD63 and PAC-1, and reduces platelet spreading. It is an inhibitor of mitochondrial NADH oxidation and electron transport in cellular respiration, it depolarizes the mitochondrial membrane, increases ROS levels, increases intracellular calcium levels and produces a decrease in ATP secretion and its intracellular levels. Western blot assays show that the antiplatelet activity induced by compound JP-I is mediated by AMPK-ACC signaling, in response to the decrease in ATP levels. Furthermore, it does not significantly alter the clot retraction (generated by Thrombin) and does not prolong the occlusion time in matrices of Collagen/Epinephrine and Collagen/ADP.
Conclusions: Our data indicate that compound JP-I decreases platelet activation by inhibiting mitochondrial bioenergetics, which activates AMPK-ACC signaling; and has a low risk of bleeding (in vitro).
To cite this abstract in AMA style:
Fuentes E, Méndez D, Palomo I, Alarcón M, Urra FA, Trostchansky A, Millas-Vargas JP, Araya-Maturana R. Antiplatelet Activity Produced by Chloroacilhidroquinones via Inhibition of the Mitochondrial Bioenergy [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/antiplatelet-activity-produced-by-chloroacilhidroquinones-via-inhibition-of-the-mitochondrial-bioenergy/. Accessed October 2, 2023.« Back to ISTH 2021 Congress
ISTH Congress Abstracts - https://abstracts.isth.org/abstract/antiplatelet-activity-produced-by-chloroacilhidroquinones-via-inhibition-of-the-mitochondrial-bioenergy/