Comparison of cytotoxic and anti-platelet activities of polyphenolic extracts from Arnica montana flowers and Juglans regia husks

Abstract Polyphenolic compounds of plant origin are well known to be beneficial to human health: they exert protective effects on haemostasis and have a particular influence on blood platelets. However, the anti-platelet properties of polyphenolic compounds observed so far have not been weighed against their potential cytotoxic action against platelets. The aim of this study was to demonstrate that anti-platelet and cytotoxic effects on blood platelets may interfere and therefore, may often lead to confusion when evaluating the properties of plant extracts or other agents towards blood platelets. The anti-platelet and cytotoxic in vitro effects of plant extracts obtained from the husks of walnuts (J. regia) and flowers of arnica (A. montana) on platelet reactivity and viability were examined. Platelet function was assessed using standard methods (flow cytometry: P-selectin expression, activation of GPIIbIIIa complex, vasodilator-stimulated phosphoprotein, VASP index; turbidimetric and impedance aggregometry) and newly set assays (flow cytometric monitoring of platelet cytotoxicity). The results reveal that none of the studied plant extracts demonstrated cytotoxicity towards blood platelets. The phenolic acid-rich extract of A. montana (7.5 and 15 µg/ml) significantly reduced the ADP-induced aggregation in both whole blood and PRP, and decreased the platelet reactivity index (PRI; VASP phosphorylation) in whole blood, while showing excellent antioxidant capacity. The extract of J. regia husks significantly reduced ADP-induced platelet aggregation in whole blood when applied at 7.5 µg/ml, and only slightly decreased the PRI at 15 µg/ml. Both examined extracts suppressed platelet hyper-reactivity, and such influence did not interfere with cytotoxic effects of the extracts. Thus, its high polyphenol content, excellent antioxidant capacity and distinct anti-platelet properties, in combination with its lack of toxicity, make the extract of A. montana flowers a possible candidate as an anti-platelet agent or a compounding diet supplement.

[1]  M. Bijak,et al.  Antithrombin Effect of Polyphenol‐Rich Extracts from Black Chokeberry and Grape Seeds , 2013, Phytotherapy research : PTR.

[2]  C. Gachet.  P2Y12 receptors in platelets and other hematopoietic and non-hematopoietic cells , 2012, Purinergic Signalling.

[3]  C. Cerletti,et al.  Effects of resveratrol and other wine polyphenols on vascular function: an update. , 2011, The Journal of nutritional biochemistry.

[4]  N. Kuhnert,et al.  Identification and characterization of five new classes of chlorogenic acids in burdock (Arctium lappa L.) roots by liquid chromatography/tandem mass spectrometry. , 2011, Food & function.

[5]  M. Ponczek,et al.  The effect of polyphenolic-polysaccharide conjugates from selected medicinal plants of Asteraceae family on the peroxynitrite-induced changes in blood platelet proteins. , 2010, International journal of biological macromolecules.

[6]  Dr David Vauzour,et al.  Polyphenols and Human Health: Prevention of Disease and Mechanisms of Action , 2010, Nutrients.

[7]  J. Pereira,et al.  Human cancer cell antiproliferative and antioxidant activities of Juglans regia L. , 2010, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[8]  P. Harrison,et al.  Markers of platelet activation and apoptosis during storage of apheresis‐ and buffy coat–derived platelet concentrates for 7 days , 2009, Transfusion.

[9]  J. Beek,et al.  Fluorescent labeling of platelets with polyanionic fluorescein derivatives. , 2009, Analytical and quantitative cytology and histology.

[10]  I. Arts,et al.  Some phenolic compounds increase the nitric oxide level in endothelial cells in vitro. , 2009, Journal of agricultural and food chemistry.

[11]  Liang Zhao,et al.  Capillary zone electrophoresis for separation and analysis of four diarylheptanoids and an alpha-tetralone derivative in the green walnut husks (Juglans regia L.). , 2008, Journal of pharmaceutical and biomedical analysis.

[12]  J. Munoz-Munoz,et al.  Kinetic characterization of the enzymatic and chemical oxidation of the catechins in green tea. , 2008, Journal of agricultural and food chemistry.

[13]  F. Violi,et al.  Resveratrol, at concentrations attainable with moderate wine consumption, stimulates human platelet nitric oxide production. , 2008, The Journal of nutrition.

[14]  L. Terry,et al.  Biochemical profiling and chemometric analysis of seventeen UK-grown black currant cultivars. , 2008, Journal of agricultural and food chemistry.

[15]  L. Estevinho,et al.  Total phenols, antioxidant potential and antimicrobial activity of walnut (Juglans regia L.) green husks. , 2008, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[16]  H. Stuppner,et al.  Quantitative analysis of flavonoids and phenolic acids in Arnica montana L. by micellar electrokinetic capillary chromatography. , 2008, Analytica chimica acta.

[17]  李辰,et al.  Capillary zone electrophoresis for separation and analysis of four diarylheptanoids and an α-tetralone derivative in the green walnut husks (Juglans regia L.) , 2008 .

[18]  Chen Li,et al.  Two new diarylheptanoids from the pericarps of Juglans regia L. , 2007 .

[19]  Chen Li,et al.  Regiolone from the pericarps of Juglans regia L. , 2007 .

[20]  D. Jacobs,et al.  Flavonoid intake and cardiovascular disease mortality: a prospective study in postmenopausal women. , 2007, The American journal of clinical nutrition.

[21]  C. Kwik-Uribe,et al.  Cocoa Flavanols and Platelet and Leukocyte Function: Recent In Vitro and Ex Vivo Studies in Healthy Adults , 2006, Journal of cardiovascular pharmacology.

[22]  F. Stampar,et al.  Traditional walnut liqueur – cocktail of phenolics , 2006 .

[23]  I. Waczulíková,et al.  High glucose contributes to aspirin insensitivity in streptozotocin-diabetic rats: a multiparametric aggregation study , 2006, Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis.

[24]  Simon B. Brown,et al.  The death of human platelets during incubation in citrated plasma involves shedding of CD42b and aggregation of dead platelets , 2005, Thrombosis and Haemostasis.

[25]  C. Gachet,et al.  Flow cytometric analysis of intraplatelet VASP phosphorylation for the detection of clopidogrel resistance in patients with ischemic cardiovascular diseases , 2005, Journal of thrombosis and haemostasis : JTH.

[26]  Rebecca E. Forkner,et al.  Determination of hydrolyzable tannins (gallotannins and ellagitannins) after reaction with potassium iodate. , 2002, Journal of agricultural and food chemistry.

[27]  M. Rozalski,et al.  Effects of fibrinogen receptor antagonist GR144053F and aurintricarboxylic acid on platelet activation and degranulation. , 2001, Biochemical pharmacology.

[28]  Henrik Stapelfeldt,et al.  Light and heat sensitivity of red cabbage extract in soft drink model systems , 2001 .

[29]  B. Campbell,et al.  Regulation of aflatoxin production by naphthoquinones of walnut (Juglans regia). , 2000, Journal of agricultural and food chemistry.

[30]  C. Rice-Evans,et al.  Antioxidant activity applying an improved ABTS radical cation decolorization assay. , 1999, Free radical biology & medicine.

[31]  J J Strain,et al.  The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay. , 1996, Analytical biochemistry.

[32]  C. Cierniewski,et al.  Rapid photometric method for estimation of platelet count in blood plasma or platelet suspension. , 1989, Thrombosis research.

[33]  G. Born,et al.  Aggregation of Blood Platelets by Adenosine Diphosphate and its Reversal , 1962, Nature.

[34]  W. Hillis,et al.  The phenolic constituents of Prunus domestica. II.†—The analysis of tissues of the Victoria plum tree , 1959 .

[35]  W. E. Hillis,et al.  The phenolic constituents of Prunus domestica. I.—The quantitative analysis of phenolic constituents , 1959 .