Characterization of a novel and potent collagen antagonist, caffeic acid phenethyl ester, in human platelets

In vitro and in vivo studies

George Hsiao, Jie J. Lee, Kuang H. Lin, Chia H. Shen, Tsorng H. Fong, Duen S. Chou, Joen R. Sheu

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Objective: Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of honeybee hives, has been demonstrated to possess multiple pharmacological activities. In the present study, CAPE (6-25 μM) specifically inhibited collagen-induced platelet aggregation and the ATP release reaction in platelet suspensions. Methods: Platelet aggregation, flow cytometric analysis, immunoblotting, and electron spin resonance (ESR) were used to assess the anti-platelet activity of CAPE. Fluorescein sodium-induced platelet thrombi in mesenteric microvessels of mice were used for an in vivo study. Results: CAPE (15-100 μM) produced a concentration-related rightward displacement of the collagen concentration-response curve, and the Schild plot gave pA2 and pA10 values of 4.28 ± 0.07 and 3.14 ± 0.73, respectively, with a slope of - 0.83 ± 0.16, indicating specific antagonism. CAPE (25 μM) also inhibited platelet aggregation stimulated by the glycoprotein VI agonist, convulxin, and the α2β1  integrin agonist, aggretin. CAPE (25 μM) also markedly interfered with FITC-collagen binding to platelet membranes. CAPE (15 and 25 μM) concentration-dependently  inhibited collagen-induced platelet activation accompanied by [Ca+2]i mobilization, phosphoinositide breakdown, activation of protein kinase C and mitogen-activated protein kinases (i.e., ERK2, JNK, and p38 MAPK), Akt phosphorylation, and thromboxane A2 formation. In the ESR study, CAPE (15 and 25 μM) markedly reduced hydroxyl radical (OH{radical dot}) formation in collagen-activated platelets. In an in vivo study, CAPE (5 mg/kg) significantly prolonged the latency in inducing platelet plug formation in mesenteric venules of mice. Conclusions: The most important findings of this study suggest that CAPE specifically inhibits collagen-induced platelet activation. Thus, CAPE treatment may represent a novel approach to lowering the risk of or improving function in thromboembolism-related disorders.

Original languageEnglish
Pages (from-to)782-792
Number of pages11
JournalCardiovascular Research
Volume75
Issue number4
DOIs
Publication statusPublished - Sep 1 2007

Fingerprint

Collagen
Blood Platelets
Platelet Aggregation
Platelet Activation
Electron Spin Resonance Spectroscopy
In Vitro Techniques
caffeic acid phenethyl ester
Propolis
Thromboxane A2
Venules
Fluorescein-5-isothiocyanate
Thromboembolism
Urticaria
p38 Mitogen-Activated Protein Kinases
Phosphatidylinositols
Microvessels
Mitogen-Activated Protein Kinases
Fluorescein
Immunoblotting
Integrins

Keywords

  • CAPE
  • Collagen antagonist
  • Hydroxyl radical
  • MAPKs
  • Platelet activation

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

@article{a831bb2f18974a3e978e47f78d934864,
title = "Characterization of a novel and potent collagen antagonist, caffeic acid phenethyl ester, in human platelets: In vitro and in vivo studies",
abstract = "Objective: Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of honeybee hives, has been demonstrated to possess multiple pharmacological activities. In the present study, CAPE (6-25 μM) specifically inhibited collagen-induced platelet aggregation and the ATP release reaction in platelet suspensions. Methods: Platelet aggregation, flow cytometric analysis, immunoblotting, and electron spin resonance (ESR) were used to assess the anti-platelet activity of CAPE. Fluorescein sodium-induced platelet thrombi in mesenteric microvessels of mice were used for an in vivo study. Results: CAPE (15-100 μM) produced a concentration-related rightward displacement of the collagen concentration-response curve, and the Schild plot gave pA2 and pA10 values of 4.28 ± 0.07 and 3.14 ± 0.73, respectively, with a slope of - 0.83 ± 0.16, indicating specific antagonism. CAPE (25 μM) also inhibited platelet aggregation stimulated by the glycoprotein VI agonist, convulxin, and the α2β1  integrin agonist, aggretin. CAPE (25 μM) also markedly interfered with FITC-collagen binding to platelet membranes. CAPE (15 and 25 μM) concentration-dependently  inhibited collagen-induced platelet activation accompanied by [Ca+2]i mobilization, phosphoinositide breakdown, activation of protein kinase C and mitogen-activated protein kinases (i.e., ERK2, JNK, and p38 MAPK), Akt phosphorylation, and thromboxane A2 formation. In the ESR study, CAPE (15 and 25 μM) markedly reduced hydroxyl radical (OH{radical dot}) formation in collagen-activated platelets. In an in vivo study, CAPE (5 mg/kg) significantly prolonged the latency in inducing platelet plug formation in mesenteric venules of mice. Conclusions: The most important findings of this study suggest that CAPE specifically inhibits collagen-induced platelet activation. Thus, CAPE treatment may represent a novel approach to lowering the risk of or improving function in thromboembolism-related disorders.",
keywords = "CAPE, Collagen antagonist, Hydroxyl radical, MAPKs, Platelet activation",
author = "George Hsiao and Lee, {Jie J.} and Lin, {Kuang H.} and Shen, {Chia H.} and Fong, {Tsorng H.} and Chou, {Duen S.} and Sheu, {Joen R.}",
year = "2007",
month = "9",
day = "1",
doi = "10.1016/j.cardiores.2007.05.005",
language = "English",
volume = "75",
pages = "782--792",
journal = "Cardiovascular Research",
issn = "0008-6363",
publisher = "Oxford University Press",
number = "4",

}

TY - JOUR

T1 - Characterization of a novel and potent collagen antagonist, caffeic acid phenethyl ester, in human platelets

T2 - In vitro and in vivo studies

AU - Hsiao, George

AU - Lee, Jie J.

AU - Lin, Kuang H.

AU - Shen, Chia H.

AU - Fong, Tsorng H.

AU - Chou, Duen S.

AU - Sheu, Joen R.

PY - 2007/9/1

Y1 - 2007/9/1

N2 - Objective: Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of honeybee hives, has been demonstrated to possess multiple pharmacological activities. In the present study, CAPE (6-25 μM) specifically inhibited collagen-induced platelet aggregation and the ATP release reaction in platelet suspensions. Methods: Platelet aggregation, flow cytometric analysis, immunoblotting, and electron spin resonance (ESR) were used to assess the anti-platelet activity of CAPE. Fluorescein sodium-induced platelet thrombi in mesenteric microvessels of mice were used for an in vivo study. Results: CAPE (15-100 μM) produced a concentration-related rightward displacement of the collagen concentration-response curve, and the Schild plot gave pA2 and pA10 values of 4.28 ± 0.07 and 3.14 ± 0.73, respectively, with a slope of - 0.83 ± 0.16, indicating specific antagonism. CAPE (25 μM) also inhibited platelet aggregation stimulated by the glycoprotein VI agonist, convulxin, and the α2β1  integrin agonist, aggretin. CAPE (25 μM) also markedly interfered with FITC-collagen binding to platelet membranes. CAPE (15 and 25 μM) concentration-dependently  inhibited collagen-induced platelet activation accompanied by [Ca+2]i mobilization, phosphoinositide breakdown, activation of protein kinase C and mitogen-activated protein kinases (i.e., ERK2, JNK, and p38 MAPK), Akt phosphorylation, and thromboxane A2 formation. In the ESR study, CAPE (15 and 25 μM) markedly reduced hydroxyl radical (OH{radical dot}) formation in collagen-activated platelets. In an in vivo study, CAPE (5 mg/kg) significantly prolonged the latency in inducing platelet plug formation in mesenteric venules of mice. Conclusions: The most important findings of this study suggest that CAPE specifically inhibits collagen-induced platelet activation. Thus, CAPE treatment may represent a novel approach to lowering the risk of or improving function in thromboembolism-related disorders.

AB - Objective: Caffeic acid phenethyl ester (CAPE), which is derived from the propolis of honeybee hives, has been demonstrated to possess multiple pharmacological activities. In the present study, CAPE (6-25 μM) specifically inhibited collagen-induced platelet aggregation and the ATP release reaction in platelet suspensions. Methods: Platelet aggregation, flow cytometric analysis, immunoblotting, and electron spin resonance (ESR) were used to assess the anti-platelet activity of CAPE. Fluorescein sodium-induced platelet thrombi in mesenteric microvessels of mice were used for an in vivo study. Results: CAPE (15-100 μM) produced a concentration-related rightward displacement of the collagen concentration-response curve, and the Schild plot gave pA2 and pA10 values of 4.28 ± 0.07 and 3.14 ± 0.73, respectively, with a slope of - 0.83 ± 0.16, indicating specific antagonism. CAPE (25 μM) also inhibited platelet aggregation stimulated by the glycoprotein VI agonist, convulxin, and the α2β1  integrin agonist, aggretin. CAPE (25 μM) also markedly interfered with FITC-collagen binding to platelet membranes. CAPE (15 and 25 μM) concentration-dependently  inhibited collagen-induced platelet activation accompanied by [Ca+2]i mobilization, phosphoinositide breakdown, activation of protein kinase C and mitogen-activated protein kinases (i.e., ERK2, JNK, and p38 MAPK), Akt phosphorylation, and thromboxane A2 formation. In the ESR study, CAPE (15 and 25 μM) markedly reduced hydroxyl radical (OH{radical dot}) formation in collagen-activated platelets. In an in vivo study, CAPE (5 mg/kg) significantly prolonged the latency in inducing platelet plug formation in mesenteric venules of mice. Conclusions: The most important findings of this study suggest that CAPE specifically inhibits collagen-induced platelet activation. Thus, CAPE treatment may represent a novel approach to lowering the risk of or improving function in thromboembolism-related disorders.

KW - CAPE

KW - Collagen antagonist

KW - Hydroxyl radical

KW - MAPKs

KW - Platelet activation

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U2 - 10.1016/j.cardiores.2007.05.005

DO - 10.1016/j.cardiores.2007.05.005

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VL - 75

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EP - 792

JO - Cardiovascular Research

JF - Cardiovascular Research

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