Luteolin inhibits GPVI-mediated platelet activation, oxidative stress, and thrombosis

Yujia Ye; Lihong Yang; Min Leng; Qian Wang; Jiankui Wu; Wen Wan; Huawei Wang; Longjun Li; Yunzhu Peng; Shengjie Chai; Zhaohui Meng

doi:10.3389/fphar.2023.1255069

Luteolin inhibits GPVI-mediated platelet activation, oxidative stress, and thrombosis

Front Pharmacol. 2023 Oct 31:14:1255069. doi: 10.3389/fphar.2023.1255069. eCollection 2023.

Authors

Yujia Ye^#¹, Lihong Yang^#¹, Min Leng^#¹, Qian Wang¹, Jiankui Wu¹, Wen Wan¹, Huawei Wang¹, Longjun Li¹, Yunzhu Peng¹, Shengjie Chai¹, Zhaohui Meng¹

Affiliation

¹ Laboratory of Molecular Cardiology, Department of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, China.

^# Contributed equally.

Abstract

Introduction: Luteolin inhibits platelet activation and thrombus formation, but the mechanisms are unclear. This study investigated the effects of luteolin on GPVI-mediated platelet activation in vitro and explored the effect of luteolin on thrombosis, coagulation, and platelet production in vivo. Methods: Washed human platelets were used for aggregation, membrane protein expression, ATP, Ca²⁺, and LDH release, platelet adhesion/spreading, and clot retraction experiments. Washed human platelets were used to detect collagen and convulxin-induced reactive oxygen species production and endogenous antioxidant effects. C57BL/6 male mice were used for ferric chloride-induced mesenteric thrombosis, collagen-epinephrine induced acute pulmonary embolism, tail bleeding, coagulation function, and luteolin toxicity experiments. The interaction between luteolin and GPVI was analyzed using solid phase binding assay and surface plasmon resonance (SPR). Results: Luteolin inhibited collagen- and convulxin-mediated platelet aggregation, adhesion, and release. Luteolin inhibited collagen- and convulxin-induced platelet ROS production and increased platelet endogenous antioxidant capacity. Luteolin reduced convulxin-induced activation of ITAM and MAPK signaling molecules. Molecular docking simulation showed that luteolin forms hydrogen bonds with GPVI. The solid phase binding assay showed that luteolin inhibited the interaction between collagen and GPVI. Surface plasmon resonance showed that luteolin bonded GPVI. Luteolin inhibited integrin αIIbβ3-mediated platelet activation. Luteolin inhibited mesenteric artery thrombosis and collagen- adrenergic-induced pulmonary thrombosis in mice. Luteolin decreased oxidative stress in vivo. Luteolin did not affect coagulation, hemostasis, or platelet production in mice. Discussion: Luteolin may be an effective and safe antiplatelet agent target for GPVI. A new mechanism (decreased oxidative stress) for the anti-platelet activity of luteolin has been identified.

Keywords: GPVI; antiplatelet; collagen; luteolin; platelet production; thrombus formation.

Grants and funding

This work was supported by the Priority Union Foundation of Yunnan Provincial Science and Technology Department and Kunming Medical University under Grant (number 202101AY070001-118); the Yunnan Fundamental Research Projects under Grant (number 202001AU070146); the First Affiliated Hospital of Kunming Medical University Doctoral Research Fund Project under Grant (number 2019BS012); the National Natural Science Foundation of China (number 82360074); and the Yunnan Provincial Clinical Medical Center of Cardio-cerebral Vascular Diseases under Grant (number ZX2019-03-01).