PB0006 - Investigation of the antiplatelet, antioxidant and anti-inflammatory potency of berries extracts

Sunday, June 25, 2023

18:30 – 19:30 ET

Room: Exhibition

Co-Author(s)

IK

Ioannis K. Koutsaliaris

Chemist, PhD Candidate
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
AT

Aikaterini N. Tsouka

Chemist, PhD Candidate
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
LP

Louisa M. Pechlivani, MSc, PhD candidate

Chemist, PhD Candidate
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
PS

Panagiotis Stathopoulos

Chemist, PhD
Division of Pharmacognosy & Natural Products Chemistry, Department of Pharmacy, NKUA, Athens, Greece
Athens, Attiki, Greece
AS

Alexandra Svouraki

Chemist, PhD
Division of Pharmacognosy & Natural Products Chemistry, Department of Pharmacy, NKUA, Athens, Greece
Athens, Attiki, Greece
CT

Constantinos Tellis

Chemist, PhD
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece
DP

Despoina Pantazi

biologist
University Hospital of Ioannina
Ioannina, Attiki, Greece
AS

Alexandros-Leandros Skaltsounis

Professorof Pharmacognosy & Natural Products Chemistry
Division of Pharmacognosy & Natural Products Chemistry, Department of Pharmacy, NKUA, Athens, Greece
Athens, Attiki, Greece

Presenting Author(s)

Alexandros D. Tselepis, MD, PhD

Emeritus Professor of Biochemistry-Clinical Chemistry, MD, PhD
Atherothrombosis Research Centre/Laboratory of Biochemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece
Ioannina, Ioannina, Greece

Background: Berries present antioxidant and anti-inflammatory properties and may exert cardioprotective activities.

Aims: We studied the effect of 2 berry extracts on the Outgrowth Endothelial Cells (OECs) activation, low-density lipoprotein (LDL) oxidative modification and platelet aggregation, in vitro.

Methods: Extraction of Morus nigra (MΝ) and Morus Alba (MA) was performed using Isopropanol/H2O (60:40v/v) and Isopropanol/H2O/Acetic Acid (60:39.5:0.5v/v/v), respectively and then treated with XAD-7 adsorbent resin, while their phytochemical profile was determined by LC-HRMS. OECs in culture were incubated with the extracts (200μg/ml) and activated with 0.5ng/mL Tumor Necrosis Factor-alpha for 6h, 37℃/5% CO2. Cell activation was determined by the membrane expression of Intercellular Adhesion Molecule-1, using flow cytometry. LDL (d=1.019-1.063g/mL) was isolated by sequential ultracentrifugation of plasma and oxidized (100μg protein/mL) by 5μM CuSO4, in the presence of extract concentrations (5-200μg/mL) for 6h, 37℃. Their inhibitory activity towards platelet aggregation, in platelet rich plasma, induced by 10μΜ ADP, 10μΜ TRAP-6 or 0.3mM Arachidonic Acid (AA) was determined by Light Transmittance Aggregometry.

Results: MN’s main compounds were anthocyanins while MA was rich in alkaloids. MN had a strong preventive effect against LDL oxidation with threshold concentration (minimum concentration with maximal inhibitory effect) of 50μg/ml and IC50 42.1±15.2, whereas MA exhibited a threshold concentration of 200μg/ml and IC50 163.6±38.9. MN and MA exhibited similar inhibitory effect in OECs activation, (~30%). MN and MA had no effect on ADP- or AA-induced platelet aggregation, whereas they showed a mild, albeit noteworthy, inhibitory effect towards TRAP-6-induced platelet aggregation (~19%).

Conclusion(s): MN had a stronger protective effect against LDL oxidation, that can be attributed to its rich anthocyanin content. The extracts exhibited similar anti-inflammatory activity and specific antiplatelet activity towards TRAP-6-induced platelet aggregation. Both extracts, primarily MN, could provide supplementary cardioprotective effects and exert an additional antiplatelet activity through the protease-activated receptor-1, in patients receiving dual antiplatelet therapy (aspirin/P2Y12 receptor antagonist).