Unveiling the mechanism of platelet dysfunction in Parkinson's disease: The effect of 6-hydroxydopamine on human blood platelets

Abstract

Introduction: Parkinson's disease (PD) is a progressive neuronal illness often linked to increased cardiovascular complications, such as myocardial infarction, cardiomyopathy, congestive heart failure, and coronary heart disease. Platelets, which are the essential components of circulating blood, are considered potential players in regulating these complications, as platelet dysfunction is evident in PD. These tiny blood cell fragments are supposed to play a crucial role in these complications, but the underlying molecular processes are still obscure.

Methods: To gain a better understanding of platelet dysfunction in PD, we investigated the impact of 6-hydroxydopamine (6-OHDA), an analog of dopamine that simulates PD by destroying dopaminergic neurons, on human blood platelets. The levels of intraplatelet reactive oxygen species (ROS) were assessed using H₂DCF-DA (20 μM), while mitochondrial ROS was evaluated using MitoSOX™ Red (5 μM), and intracellular Ca²⁺ was measured with Fluo-4-AM (5 μM). The data were acquired through the use of both a multimode plate reader and a laser-scanning confocal microscope.

Results: Our findings showed that 6-OHDA treatment increased the production of ROS in human blood platelets. The increase in ROS was confirmed by the ROS scavenger, NAC, and was also reduced by inhibiting the NOX enzyme with apocynin. Additionally, 6-OHDA potentiated mitochondrial ROS production in platelets. Furthermore, 6-OHDA triggered the intraplatelet Ca²⁺ elevation. This effect was mitigated by the Ca²⁺ chelator BAPTA, which decreased the ROS production triggered by 6-OHDA in human blood platelets, while the IP₃ receptor blocker, 2-APB, reduced the formation of ROS induced by 6-OHDA.

Conclusion: Our findings suggest that the 6-OHDA-induced ROS production is regulated by the IP₃ receptor-Ca²⁺-NOX signaling axis in human blood platelets, where the platelet mitochondria also play a significant role. This observation provides a crucial mechanistic understanding of the altered platelet activities that are commonly observed in PD patients.

Keywords: 6-Hydroxydopamine; Blood platelet; IP(3) receptor; Intracellular Ca(2+); Mitochondrial ROS; NADPH oxidase; Parkinson's disease.