Plant-based Food Cyanidin-3-Glucoside Modulates Human Platelet Glycoprotein VI Signaling and Inhibits Platelet Activation and Thrombus Formation

J Nutr. 2017 Oct;147(10):1917-1925. doi: 10.3945/jn.116.245944. Epub 2017 Aug 30.

Abstract

Background: Platelets play an important role in hemostasis, thrombosis, and atherosclerosis. Glycoprotein VI (GPVI) is a major platelet receptor that interacts with exposed collagen on injured vessel walls. Our previous studies have shown that anthocyanins (a type of natural plant pigment) attenuate platelet function; however, whether anthocyanins affect collagen-induced GPVI signaling remains unknown.Objective: The objective of this study was to explore the effects of cyanidin-3-glucoside (Cy-3-g, one of the major bioactive compounds in anthocyanins) on platelet activation and thrombosis and the GPVI signaling pathway.Methods: Platelets from healthy men and women were isolated and incubated with different concentrations (0, 0.5, 5, and 50 μM) of Cy-3-g. The expression of activated integrin αIIbβ3, P-selectin, CD63, and CD40L, fibrinogen binding to platelets, and platelet aggregation were evaluated in vitro. Platelet adhesion and aggregation in whole blood under flow conditions were assessed in collagen-coated perfusion chambers. Thrombosis and hemostasis were assessed in 3-4-wk-old male C57BL/6J mice through FeCl3-induced intravital microscopy and tail bleeding time. The effect of Cy-3-g on collagen-induced human platelet GPVI signaling was explored with Western blot.Results: Cy-3-g attenuated platelet function in a dose-dependent manner. The 0.5-μM dose of Cy-3-g inhibited (P < 0.05) human platelet adhesion and aggregation to collagen at both venous (-54.02%) and arterial (-22.90%) shear stresses. The 5-μM dose inhibited (P < 0.05) collagen-induced human platelet activation (PAC-1: -48.21%, P-selectin: -50.63%), secretion (CD63: -73.89%, CD40L: -43.70%), fibrinogen binding (-56.79%), and aggregation (-17.81%). The 5-μM dose attenuated (P < 0.01) thrombus growth (-66.67%) without prolonging bleeding time in mice. The 50-μM dose downregulated (P < 0.05) collagen-induced GPVI signaling in human platelets and significantly decreased phosphorylation of Syk-linker for activation of T cells (LAT)-SLP76 (Syk: -39.08%, LAT: -32.25%, SLP76: -40.00%) and the expression of Lyn (-31.89%), Fyn (-36.27%), and phospholipase C-γ2 (-39.08%).Conclusions: Cy-3-g inhibits human platelet activation, aggregation, secretion, and thrombus formation, and downregulates the collagen-GPVI signaling pathway. Supplementation of Cy-3-g may have protective effects against atherothrombosis.

Keywords: CD40L; CD63; Cy-3-g; GPVI; P-selectin; anthocyanins; collagen; platelets; thrombosis and hemostasis; αIIbβ3 integrin.

MeSH terms

  • Adaptor Proteins, Signal Transducing / blood
  • Adult
  • Aged
  • Animals
  • Anthocyanins / pharmacology
  • Anthocyanins / therapeutic use
  • Antigens, CD / blood
  • Atherosclerosis / blood
  • Atherosclerosis / diet therapy
  • Atherosclerosis / etiology
  • Blood Platelets / metabolism*
  • Collagen / blood
  • Female
  • Glucosides / pharmacology
  • Glucosides / therapeutic use
  • Hemostasis / drug effects
  • Humans
  • Male
  • Mice, Inbred C57BL
  • Middle Aged
  • P-Selectin / blood
  • Phosphoproteins / blood
  • Phytotherapy*
  • Plant Extracts / pharmacology*
  • Plant Extracts / therapeutic use
  • Plants, Edible / chemistry*
  • Platelet Activation / drug effects
  • Platelet Aggregation / drug effects*
  • Platelet Membrane Glycoproteins / metabolism*
  • Signal Transduction
  • Thrombosis / blood
  • Thrombosis / etiology
  • Thrombosis / prevention & control*

Substances

  • Adaptor Proteins, Signal Transducing
  • Anthocyanins
  • Antigens, CD
  • Glucosides
  • P-Selectin
  • Phosphoproteins
  • Plant Extracts
  • Platelet Membrane Glycoproteins
  • SELP protein, human
  • SLP-76 signal Transducing adaptor proteins
  • cyanidin-3-O-beta-glucopyranoside
  • platelet membrane glycoprotein VI
  • Collagen