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Thromb Haemost 2021; 121(07): 931-943
DOI: 10.1055/s-0040-1722621
Cellular Haemostasis and Platelets

Protocatechuic Acid Protects Platelets from Apoptosis via Inhibiting Oxidative Stress-Mediated PI3K/Akt/GSK3β Signaling

Fuli Ya‡   *
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
,
Kongyao Li‡   *
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
,
Hong Chen
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
,
Zezhong Tian
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
,
Die Fan
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
,
Yilin Shi
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
4   Department of Nutrition, School of Public Health (Northern Campus), Sun Yat-sen University, Guangzhou, Guangdong Province, China
,
Fenglin Song
5   Department of Food Safety, School of Food Science, Guangdong Pharmaceutical University, Guangzhou, Guangdong Province, China
,
Xiping Xu
6   Renal Division, National Clinical Research Center for Kidney Disease, Southern Medical University, Nanfang Hospital, Guangzhou, Guangdong Province, China
,
Wenhua Ling
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
4   Department of Nutrition, School of Public Health (Northern Campus), Sun Yat-sen University, Guangzhou, Guangdong Province, China
,
Reheman Adili
7   Department of Pharmacology, University of Michigan, Ann Arbor, Michigan, United States
,
Yan Yang
1   Department of Nutrition and Food Safety, School of Public Health (Shenzhen), Sun Yat-sen University, Guangzhou, Guangdong Province, China
2   Guangdong Provincial Key Laboratory for Food, Nutrition and Health, Guangzhou, Guangdong Province, China
3   Guangdong Engineering Technology Research Center of Nutrition Translation, Guangzhou, Guangdong Province, China
› Author Affiliations Funding This work was supported by the Guangzhou Science, Technology, and Innovation Commission (No. 201804020045), the Key Project of National Natural Science Foundation of China (No. 81730090 and No. 82030098), the National Natural Science Foundation of China (No. 81872617), and the Shenzhen Science and Technology Innovation Commission (No. 201803073000433).
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Abstract

Oxidative stress plays crucial roles in initiating platelet apoptosis that facilitates the progression of cardiovascular diseases (CVDs). Protocatechuic acid (PCA), a major metabolite of anthocyanin cyanidin-3-O-β-glucoside (Cy-3-g), exerts cardioprotective effects. However, underlying mechanisms responsible for such effects remain unclear. Here, we investigate the effect of PCA on platelet apoptosis and the underlying mechanisms in vitro. Isolated human platelets were treated with hydrogen peroxide (H2O2) to induce apoptosis with or without pretreatment with PCA. We found that PCA dose-dependently inhibited H2O2-induced platelet apoptosis by decreasing the dissipation of mitochondrial membrane potential, activation of caspase-9 and caspase-3, and decreasing phosphatidylserine exposure. Additionally, the distributions of Bax, Bcl-xL, and cytochrome c mediated by H2O2 in the mitochondria and the cytosol were also modulated by PCA treatment. Moreover, the inhibitory effects of PCA on platelet caspase-3 cleavage and phosphatidylserine exposure were mainly mediated by downregulating PI3K/Akt/GSK3β signaling. Furthermore, PCA dose-dependently decreased reactive oxygen species (ROS) generation and the intracellular Ca2+ concentration in platelets in response to H2O2. N-Acetyl cysteine (NAC), a ROS scavenger, markedly abolished H2O2-stimulated PI3K/Akt/GSK3β signaling, caspase-3 activation, and phosphatidylserine exposure. The combination of NAC and PCA did not show significant additive inhibitory effects on PI3K/Akt/GSK3β signaling and platelet apoptosis. Thus, our results suggest that PCA protects platelets from oxidative stress-induced apoptosis through downregulating ROS-mediated PI3K/Akt/GSK3β signaling, which may be responsible for cardioprotective roles of PCA in CVDs.

Keywords

anthocyanin - protocatechuic acid - platelet apoptosis - reactive oxygen species - oxidative stress

* These authors contributed equally to this work.


Supplementary Material



Publication History

Received: 21 December 2019

Accepted: 30 November 2020

Article published online:
05 February 2021

© 2021. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Xu XR, Zhang D, Oswald BE. et al. Platelets are versatile cells: new discoveries in hemostasis, thrombosis, immune responses, tumor metastasis and beyond. Crit Rev Clin Lab Sci 2016; 53 (06) 409-430
    CrossrefPubMedGoogle Scholar
  • 2 Thushara RM, Hemshekhar M, , Basappa, Kemparaju K, Rangappa KS, Girish KS. Biologicals, platelet apoptosis and human diseases: An outlook. Crit Rev Oncol Hematol 2015; 93 (03) 149-158
    CrossrefPubMedGoogle Scholar
  • 3 Pietraforte D, Vona R, Marchesi A. et al. Redox control of platelet functions in physiology and pathophysiology. Antioxid Redox Signal 2014; 21 (01) 177-193
    CrossrefPubMedGoogle Scholar
  • 4 Leytin V. Apoptosis in the anucleate platelet. Blood Rev 2012; 26 (02) 51-63
    CrossrefPubMedGoogle Scholar
  • 5 Kile BT. The role of the intrinsic apoptosis pathway in platelet life and death. J Thromb Haemost 2009; 7 (Suppl. 01) 214-217
    CrossrefPubMedGoogle Scholar
  • 6 Wojta J. Platelet-derived microparticles in patients with high cardiovascular risk and subclinical atherosclerosis. Thromb Haemost 2015; 114 (06) 1099
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Wang D, Xia M, Yan X. et al. Gut microbiota metabolism of anthocyanin promotes reverse cholesterol transport in mice via repressing miRNA-10b. Circ Res 2012; 111 (08) 967-981
    CrossrefPubMedGoogle Scholar
  • 8 Ya F, Xu XR, Tian Z. et al. Coenzyme Q10 attenuates platelet integrin αIIbβ3 signaling and platelet hyper-reactivity in ApoE-deficient mice. Food Funct 2020; 11 (01) 139-152
    CrossrefPubMedGoogle Scholar
  • 9 He J, Giusti MM. Anthocyanins: natural colorants with health-promoting properties. Annu Rev Food Sci Technol 2010; 1: 163-187
    CrossrefPubMedGoogle Scholar
  • 10 Song F, Zhu Y, Shi Z. et al. Plant food anthocyanins inhibit platelet granule secretion in hypercholesterolaemia: involving the signalling pathway of PI3K-Akt. Thromb Haemost 2014; 112 (05) 981-991
    Article in Thieme ConnectPubMedGoogle Scholar
  • 11 Zhang X, Zhu Y, Song F. et al. Effects of purified anthocyanin supplementation on platelet chemokines in hypocholesterolemic individuals: a randomized controlled trial. Nutr Metab (Lond) 2016; 13: 86
    CrossrefPubMedGoogle Scholar
  • 12 Huang W, Zhu Y, Li C, Sui Z, Min W. Effect of blueberry anthocyanins malvidin and glycosides on the antioxidant properties in endothelial cells. Oxid Med Cell Longev 2016; 2016: 1591803
    CrossrefPubMedGoogle Scholar
  • 13 Huang WY, Liu YM, Wang J, Wang XN, Li CY. Anti-inflammatory effect of the blueberry anthocyanins malvidin-3-glucoside and malvidin-3-galactoside in endothelial cells. Molecules 2014; 19 (08) 12827-12841
    CrossrefPubMedGoogle Scholar
  • 14 Yao Y, Chen Y, Adili R. et al. Plant-based food cyanidin-3-glucoside modulates human platelet glycoprotein VI signaling and inhibits platelet activation and thrombus formation. J Nutr 2017; 147 (10) 1917-1925
    CrossrefPubMedGoogle Scholar
  • 15 Zhou FH, Deng XJ, Chen YQ. et al. Anthocyanin cyanidin-3-glucoside attenuates platelet granule release in mice fed high-fat diets. J Nutr Sci Vitaminol (Tokyo) 2017; 63 (04) 237-243
    CrossrefPubMedGoogle Scholar
  • 16 Ya F, Li Q, Wang D. et al. Cyanidin-3-o-β-glucoside induces megakaryocyte apoptosis via PI3K/Akt- and MAPKs-mediated inhibition of NF-κB signalling. Thromb Haemost 2018; 118 (07) 1215-1229
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Ya F, Tian J, Li Q. et al. Cyanidin-3-O-β-glucoside, a natural polyphenol, exerts proapoptotic effects on activated platelets and enhances megakaryocytic proplatelet formation. J Agric Food Chem 2018; 66 (41) 10712-10720
    CrossrefPubMedGoogle Scholar
  • 18 Czank C, Cassidy A, Zhang Q. et al. Human metabolism and elimination of the anthocyanin, cyanidin-3-glucoside: a (13)C-tracer study. Am J Clin Nutr 2013; 97 (05) 995-1003
    CrossrefPubMedGoogle Scholar
  • 19 Kim K, Bae ON, Lim KM. et al. Novel antiplatelet activity of protocatechuic acid through the inhibition of high shear stress-induced platelet aggregation. J Pharmacol Exp Ther 2012; 343 (03) 704-711
    CrossrefPubMedGoogle Scholar
  • 20 Krga I, Vidovic N, Milenkovic D. et al. Effects of anthocyanins and their gut metabolites on adenosine diphosphate-induced platelet activation and their aggregation with monocytes and neutrophils. Arch Biochem Biophys 2018; 645: 34-41
    CrossrefPubMedGoogle Scholar
  • 21 Ya F, Xu XR, Shi Y. et al. Coenzyme Q10 upregulates platelet cAMP/PKA pathway and attenuates integrin αIIbβ3 signaling and thrombus growth. Mol Nutr Food Res 2019; 63 (23) e1900662
    CrossrefPubMedGoogle Scholar
  • 22 Xu XR, Wang Y, Adili R. et al. Apolipoprotein A-IV binds αIIbβ3 integrin and inhibits thrombosis. Nat Commun 2018; 9 (01) 3608
    CrossrefPubMedGoogle Scholar
  • 23 Tang WH, Stitham J, Jin Y. et al. Aldose reductase-mediated phosphorylation of p53 leads to mitochondrial dysfunction and damage in diabetic platelets. Circulation 2014; 129 (15) 1598-1609
    CrossrefPubMedGoogle Scholar
  • 24 Wei H, Harper MT. ABT-737 triggers caspase-dependent inhibition of platelet procoagulant extracellular vesicle release during apoptosis and secondary necrosis in vitro. Thromb Haemost 2019; 119 (10) 1665-1674
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Suades R, Padró T, Vilahur G, Badimon L. Circulating and platelet-derived microparticles in human blood enhance thrombosis on atherosclerotic plaques. Thromb Haemost 2012; 108 (06) 1208-1219
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Ridger VC, Boulanger CM, Angelillo-Scherrer A. et al; Position Paper of the European Society of Cardiology (ESC) Working Group on Atherosclerosis and Vascular Biology. Microvesicles in vascular homeostasis and diseases. Thromb Haemost 2017; 117 (07) 1296-1316
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Krga I, Milenkovic D. Anthocyanins: from sources and bioavailability to cardiovascular-health benefits and molecular mechanisms of action. J Agric Food Chem 2019; 67 (07) 1771-1783
    CrossrefPubMedGoogle Scholar
  • 28 Pang A, Cui Y, Chen Y. et al. Shear-induced integrin signaling in platelet phosphatidylserine exposure, microvesicle release, and coagulation. Blood 2018; 132 (05) 533-543
    CrossrefPubMedGoogle Scholar
  • 29 Gyulkhandanyan AV, Mutlu A, Freedman J, Leytin V. Markers of platelet apoptosis: methodology and applications. J Thromb Thrombolysis 2012; 33 (04) 397-411
    CrossrefPubMedGoogle Scholar
  • 30 Sener A, Ozsavci D, Oba R, Demirel GY, Uras F, Yardimci KT. Do platelet apoptosis, activation, aggregation, lipid peroxidation and platelet-leukocyte aggregate formation occur simultaneously in hyperlipidemia?. Clin Biochem 2005; 38 (12) 1081-1087
    CrossrefPubMedGoogle Scholar
  • 31 Owens III AP, Mackman N. Microparticles in hemostasis and thrombosis. Circ Res 2011; 108 (10) 1284-1297
    CrossrefPubMedGoogle Scholar
  • 32 Durante A, Zalewski J. Microparticles: a novel player in cardiovascular diseases. Cardiology 2015; 132 (04) 249-251
    CrossrefPubMedGoogle Scholar
  • 33 Burnouf T, Goubran HA, Chou ML, Devos D, Radosevic M. Platelet microparticles: detection and assessment of their paradoxical functional roles in disease and regenerative medicine. Blood Rev 2014; 28 (04) 155-166
    CrossrefPubMedGoogle Scholar
  • 34 Podrez EA, Byzova TV, Febbraio M. et al. Platelet CD36 links hyperlipidemia, oxidant stress and a prothrombotic phenotype. Nat Med 2007; 13 (09) 1086-1095
    CrossrefPubMedGoogle Scholar
  • 35 Lee SH, Du J, Stitham J. et al. Inducing mitophagy in diabetic platelets protects against severe oxidative stress. EMBO Mol Med 2016; 8 (07) 779-795
    CrossrefPubMedGoogle Scholar
  • 36 Freedman JE. Oxidative stress and platelets. Arterioscler Thromb Vasc Biol 2008; 28 (03) s11-s16
    CrossrefPubMedGoogle Scholar
  • 37 Pignatelli P, Pulcinelli FM, Lenti L, Gazzaniga PP, Violi F. Hydrogen peroxide is involved in collagen-induced platelet activation. Blood 1998; 91 (02) 484-490
    CrossrefPubMedGoogle Scholar
  • 38 Han L, Yang Q, Ma W, Li J, Qu L, Wang M. Protocatechuic acid ameliorated palmitic-acid-induced oxidative damage in endothelial cells through activating endogenous antioxidant enzymes via an adenosine-monophosphate-activated-protein-kinase-dependent pathway. J Agric Food Chem 2018; 66 (40) 10400-10409
    CrossrefPubMedGoogle Scholar
  • 39 Bhattacharjee N, Dua TK, Khanra R. et al. Protocatechuic acid, a phenolic from Sansevieria roxburghiana leaves, suppresses diabetic cardiomyopathy via stimulating glucose metabolism, ameliorating oxidative stress, and inhibiting inflammation. Front Pharmacol 2017; 8: 251
    CrossrefPubMedGoogle Scholar
  • 40 Adedara IA, Omole O, Okpara ES. et al. Impact of prepubertal exposure to dietary protocatechuic acid on the hypothalamic-pituitary-testicular axis in rats. Chem Biol Interact 2018; 290: 99-109
    CrossrefPubMedGoogle Scholar
  • 41 O'Brien KA, Gartner TK, Hay N, Du X. ADP-stimulated activation of Akt during integrin outside-in signaling promotes platelet spreading by inhibiting glycogen synthase kinase-3β. Arterioscler Thromb Vasc Biol 2012; 32 (09) 2232-2240
    CrossrefPubMedGoogle Scholar
  • 42 Laurent PA, Severin S, Gratacap MP, Payrastre B. Class I PI 3-kinases signaling in platelet activation and thrombosis: PDK1/Akt/GSK3 axis and impact of PTEN and SHIP1. Adv Biol Regul 2014; 54: 162-174
    CrossrefPubMedGoogle Scholar
  • 43 Chen M, Yan R, Zhou K. et al. Akt-mediated platelet apoptosis and its therapeutic implications in immune thrombocytopenia. Proc Natl Acad Sci U S A 2018; 115 (45) E10682-E10691
    CrossrefPubMedGoogle Scholar
  • 44 Rosado JA, Redondo PC, Salido GM, Gómez-Arteta E, Sage SO, Pariente JA. Hydrogen peroxide generation induces pp60src activation in human platelets: evidence for the involvement of this pathway in store-mediated calcium entry. J Biol Chem 2004; 279 (03) 1665-1675
    CrossrefPubMedGoogle Scholar
  • 45 Li Z, Delaney MK, O'Brien KA, Du X. Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol 2010; 30 (12) 2341-2349
    CrossrefPubMedGoogle Scholar
  • 46 Praticò D, Iuliano L, Ghiselli A, Alessandri C, Violi F. Hydrogen peroxide as trigger of platelet aggregation. Haemostasis 1991; 21 (03) 169-174
    PubMedGoogle Scholar
  • 47 Ma Y, Chen F, Yang S, Chen B, Shi J. Protocatechuic acid ameliorates high glucose-induced extracellular matrix accumulation in diabetic nephropathy. Biomed Pharmacother 2018; 98: 18-22
    CrossrefPubMedGoogle Scholar
  • 48 Song H, Ren J. Protocatechuic acid attenuates angiotensin II-induced cardiac fibrosis in cardiac fibroblasts through inhibiting the NOX4/ROS/p38 signaling pathway. Phytother Res 2019; 33 (09) 2440-2447
    CrossrefPubMedGoogle Scholar
  • 49 Yang M, Cooley BC, Li W. et al. Platelet CD36 promotes thrombosis by activating redox sensor ERK5 in hyperlipidemic conditions. Blood 2017; 129 (21) 2917-2927
    CrossrefPubMedGoogle Scholar
  • 50 Magwenzi S, Woodward C, Wraith KS. et al. Oxidized LDL activates blood platelets through CD36/NOX2-mediated inhibition of the cGMP/protein kinase G signaling cascade. Blood 2015; 125 (17) 2693-2703
    CrossrefPubMedGoogle Scholar