A Novel Role of Eruca sativa Mill. (rocket) extract: Antiplatelet (NF-κB Inhibition) and antithrombotic activities

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Abstract

Background: Epidemiological studies have shown the prevention of cardiovascular diseases through the regular consumption of vegetables. Eruca sativa Mill., commonly known as rocket, is a leafy vegetable that has anti-inflammatory activity. However, its antiplatelet and antithrombotic activities have not been described. Methods: Eruca sativa Mill. aqueous extract (0.1 to 1 mg/mL), was evaluated on human platelets: (i) P-selectin expression by flow cytometry; (ii) platelet aggregation induced by ADP, collagen and arachidonic acid; (iii) IL-1β, TGF-β1, CCL5 and thromboxane B2 release; and (iv) activation of NF-κB and PKA by western blot. Furthermore, (v) antithrombotic activity (200 mg/kg) and (vi) bleeding time in murine models were evaluated. Results: Eruca sativa Mill. aqueous extract (0.1 to 1 mg/mL) inhibited P-selectin expression and platelet aggregation induced by ADP. The release of platelet inflammatory mediators (IL-1β, TGF-β1, CCL5 and thromboxane B2) induced by ADP was inhibited by Eruca sativa Mill. aqueous extract. Furthermore, Eruca sativa Mill. aqueous extract inhibited NF-κB activation. Finally, in murine models, Eruca sativa Mill. aqueous extract showed significant antithrombotic activity and a slight effect on bleeding time. Conclusion: Eruca sativa Mill. presents antiplatelet and antithrombotic activity.

Figures

  • Figure 1. Effect of Eruca sativa Mill. extract on platelet activation. P-selectin expression was determined by flow cytometry. The basal bar represents the fluorescence in the non-stimulated sample. The graph depicts the mean ± SEM of n = 6 experiments. An asterisk (* p < 0.05) denotes a statistically significant difference when Eruca sativa Mill. extract was compared with the activated control analyzed by one-way ANOVA and Tukey’s post hoc test.
  • Figure 2. Effects of Eruca sativa Mill. extract on ADP (8 μmol/L), collagen (1.5 µg/mL) and AA (1 mmol/L) induced platelet aggregation. Results were expressed as platelet aggregation (%) (mean ± SEM, n = 6). Asterisks (* p < 0.05 and *** p < 0.001) denote a statistically significant difference when compared with the activated control analyzed by one-way ANOVA and Tukey’s post hoc test. AA = arachidonic acid.
  • Figure 3. Effects of Eruca sativa Mill. extract on the release of thromboxane B2 (A), CCL5 (B), TGF-β1 (C) and IL-1β (D) induced by ADP (8 μmol/L) from human platelets. The graph depicts the mean ± SEM of n = 6 experiments. ** p < 0.01 and *** p < 0.001 indicates differences when compared with activated control, analyzed by one-way ANOVA and Tukey’s post hoc test.
  • Figure 4. Effect of Eruca sativa Mill. extract on phospho-PKA in activated platelets by ADP (8 μmol/L). The activated group corresponds to prostaglandin E1 (PGE1) plus ADP. Data are presented as the mean ± SEM of n = 6 experiments. *** p < 0.001 indicates the difference between activated and basal groups as analyzed by one-way ANOVA and Tukey’s post hoc test.
  • Figure 5. Effect of Eruca sativa Mill. extract on phospho-NF-κB p65 in activated platelets by ADP (8 μmol/L). Washed platelets were collected, and subcellular extracts were analyzed for phospho-NF-κB p65, as described in the Experimental Section. Data are presented as the mean ± SEM of n = 6 experiments. ** p < 0.01 and *** p < 0.001 indicates differences between activated and Eruca sativa Mill. extract groups as analyzed by one-way ANOVA and Tukey’s post hoc test.
  • Figure 6. Eruca sativa Mill. extract inhibited arterial thrombosis formation. Representative images of thrombus formation after laser irradiation in the vehicle (DMSO 0.2%) control group (n = 6), ASA (acetylsalicylic acid; 200 mg/kg; n = 6) and Eruca sativa extract (200 mg/kg, n = 6) to 60 min. Each photograph represents the percentage of occlusions caused by thrombus formation. I, intima; M, media; A, adventitia.

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Fuentes, E., Alarcón, M., Fuentes, M., Carrasco, G., & Palomo, I. (2014). A Novel Role of Eruca sativa Mill. (rocket) extract: Antiplatelet (NF-κB Inhibition) and antithrombotic activities. Nutrients, 6(12), 5839–5852. https://doi.org/10.3390/nu6125839

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