Gauer Julia S, Ajanel Abigail, Kaselampao Lutale M, Candir Isabel, MacCannell Amanda D V, Roberts Lee D, Campbell Robert A, Ariëns Robert A S
Discovery and Translational Science Department, Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom.
Molecular Medicine Program, University of Utah, Salt Lake City, UT 84112, USA.
Res Pract Thromb Haemost. 2024 Aug 14;8(6):102548. doi: 10.1016/j.rpth.2024.102548. eCollection 2024 Aug.
Polyphenols have been shown to decrease oxidative stress and modulate glycemic response. Nevertheless, their effect on platelet bioenergetics and clot structure in diabetes and hyperglycemia is unknown.
To investigate the effect of polyphenols on human platelet bioenergetics and its subsequent effect on clot structure in normoglycemia vs acute hyperglycemia .
Four polyphenols (resveratrol, hesperetin, epigallocatechin gallate [EGCG], and quercetin) were selected for initial analysis. Healthy volunteers' isolated platelets/platelet-rich plasma were treated with 5 or 25 mM glucose to represent normoglycemia and acute hyperglycemia, respectively. Platelet-derived reactive oxygen species (ROS), citrate synthase activity (mitochondrial density), mitochondrial calcium flux, and mitochondrial respiration were performed following exposure to polyphenols (20 µM, 1 hour) to determine their effects on platelet bioenergetics. Procoagulant platelets (annexin V) and fibrin fiber density (Alexa Fluor-488 fibrinogen; Invitrogen) were analyzed by laser scanning confocal microscopy, while clot porosity was determined using platelet-rich plasma following exposure to polyphenols (20 µM, 20 minutes).
Acute hyperglycemia increased ROS, mitochondrial calcium flux, maximal respiration, and procoagulant platelet number. Resveratrol, quercetin, and EGCG reduced platelet ROS in normoglycemic and acute hyperglycemic conditions. Mitochondrial density was decreased by quercetin and EGCG in normoglycemia. Resveratrol and EGCG reduced mitochondrial calcium flux in acute hyperglycemia. Resveratrol also decreased procoagulant platelet number and attenuated oxygen consumption rate in normoglycemia and acute hyperglycemia. No effect of hyperglycemia or polyphenols was observed on fibrin fiber density or clot pore size.
Our results suggest polyphenols attenuate increased platelet activity stemming from hyperglycemia and may benefit thrombosis-preventative strategies in patients with diabetes.
多酚已被证明可降低氧化应激并调节血糖反应。然而，它们对糖尿病和高血糖状态下血小板生物能量学及血凝块结构的影响尚不清楚。
研究多酚对正常血糖与急性高血糖状态下人体血小板生物能量学的影响及其对血凝块结构的后续作用。
选择四种多酚（白藜芦醇、橙皮素、表没食子儿茶素没食子酸酯[EGCG]和槲皮素）进行初步分析。将健康志愿者分离出的血小板/富含血小板血浆分别用5 mM或25 mM葡萄糖处理，以分别代表正常血糖和急性高血糖状态。在暴露于多酚（20 μM，1小时）后，检测血小板衍生的活性氧（ROS）、柠檬酸合酶活性（线粒体密度）、线粒体钙通量和线粒体呼吸，以确定它们对血小板生物能量学的影响。通过激光扫描共聚焦显微镜分析促凝血血小板（膜联蛋白V）和纤维蛋白纤维密度（Alexa Fluor - 488纤维蛋白原；Invitrogen公司），同时在暴露于多酚（20 μM，20分钟）后，使用富含血小板血浆测定血凝块孔隙率。
急性高血糖增加了ROS、线粒体钙通量、最大呼吸和促凝血血小板数量。白藜芦醇、槲皮素和EGCG在正常血糖和急性高血糖条件下均降低了血小板ROS。在正常血糖状态下，槲皮素和EGCG降低了线粒体密度。白藜芦醇和EGCG在急性高血糖状态下降低了线粒体钙通量。白藜芦醇在正常血糖和急性高血糖状态下还降低了促凝血血小板数量并减弱了氧消耗率。未观察到高血糖或多酚对纤维蛋白纤维密度或血凝块孔径有影响。
我们的结果表明，多酚可减轻高血糖引起的血小板活性增加，可能对糖尿病患者的血栓预防策略有益。
