Key Laboratory of Atherosclerosis in Universities of Shandong, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian 271000, PR China.
Province Key Laboratory of Oral and Maxillofacial, Head and Neck Medical Biology Laboratory, Liaocheng People's Hospital, Shandong First Medical University & Shandong Academy of Medical Sciences, Liaocheng 252000, PR China.
Life Sci. 2019 Sep 15;233:116700. doi: 10.1016/j.lfs.2019.116700. Epub 2019 Jul 26.
Hydrogen (H) has antioxidant effects. The pharmacologic function of H in platelets is not yet clear. Therefore, in this study we sought to investigate the inhibitory effects of H on in vitro platelet activation and in vivo prevention of thrombus formation.
After platelets were incubated with H-rich saline (HRS), platelet adhesion in whole human blood was assessed in fibrinogen-coated perfusion chambers, while rat platelet aggregation induced by ADP, collagen and HO was detected through light transmission aggregometry. The level of P-selectin, thromboxane B, nitric oxide (NO), malondialdehyde, reactive oxygen species (ROS), cGMP, extracellular signal-regulated kinases 1 and 2 (p-ERK1/2), and fibrinogen binding to platelets were evaluated in vitro. Besides, the in vivo effects were examined in arterio-venous shunt thrombosis, FeCl-induced artery thrombus formation, and tail bleeding time in mice and rats.
HRS prolonged tail bleeding time in mice and rats, decreased thrombus weight and prolonged the time to occlusion in rat and mouse thrombosis models in vivo and inhibited platelet adhesion as well as aggregation in vitro. Additionally, HRS decreased P-selectin expression, release of thromboxane B, ROS, and fibrinogen binding, but enhanced NO levels in HO-exposed platelets. HRS also decreased malondialdehyde levels in plasma of the rat arterial thrombosis or HO-exposed platelet model. Moreover, HRS increased cGMP level, decreased p-ERK1/2 (diminished with KT5823) in the platelets stimulated by HO.
These results suggest that H has antithrombotic effects, which may be due to its antioxidant property and subsequent inhibition of platelet activation via NO/cGMP/PKG/ERK pathway.
氢气（H）具有抗氧化作用。H 在血小板中的药理作用尚不清楚。因此，在这项研究中，我们试图研究 H 对体外血小板活化的抑制作用和体内血栓形成的预防作用。
将富含 H 的生理盐水（HRS）孵育血小板后，在纤维蛋白原包被的灌注室中评估全人血中血小板的黏附，通过透光比浊法检测 ADP、胶原和 HO 诱导的大鼠血小板聚集。体外评估 P-选择素、血栓素 B、一氧化氮（NO）、丙二醛、活性氧（ROS）、环鸟苷酸（cGMP）、细胞外信号调节激酶 1 和 2（p-ERK1/2）的水平以及血小板与纤维蛋白原的结合。此外，在动静脉分流血栓形成、FeCl 诱导的动脉血栓形成和小鼠和大鼠的尾出血时间中检测体内作用。
HRS 延长了小鼠和大鼠的尾出血时间，减少了血栓重量，并延长了体内大鼠和小鼠血栓模型的闭塞时间，抑制了体外血小板黏附和聚集。此外，HRS 降低了 P-选择素表达、血栓素 B、ROS 和纤维蛋白原结合的释放，但增强了 HO 暴露血小板中的 NO 水平。HRS 还降低了血浆中丙二醛水平动脉血栓形成或 HO 暴露血小板模型。此外，HRS 增加了 cGMP 水平，降低了 HO 刺激血小板中 p-ERK1/2（用 KT5823 减弱）。
这些结果表明 H 具有抗血栓形成作用，这可能是由于其抗氧化特性以及随后通过 NO/cGMP/PKG/ERK 途径抑制血小板活化。
