INVESTIGADORES
SABBIONE Ana Clara
congresos y reuniones científicas
Título:
Amaranth peptides decreased the activity and expression of cellular tissue factor on LPS activated THP-1 human monocytes
Autor/es:
ANA CLARA SABBIONE; DIEGO ARMANDO LUNA-VITAL; MARÍA CRISTINA AÑÓN; MARÍA ELVIRA GONZÁLEZ DE MEJÍA
Lugar:
Boston
Reunión:
Congreso; American Society for Nutrition, Nutrition 2018; 2018
Resumen:
Objectives: the objective of the present research was to evaluate the antithrombotic and anti-inflammatory effect of amaranth peptides over tissue factor (TF) activity and expression on THP-1 activated cells.Methods: Amaranthus mantegazzianus protein isolates were obtained through alkaline precipitation and simulated gastrointestinal digestion of amaranth protein isolates(SGD) was performed using pepsin and pancreatin pool of enzymes. An active anticoagulant peptide fraction (AF) was acquired using FPLC chromatography. Human THP-1 monocytes were treated for 24 h with different concentrations of sterile-filtered SGD and AF, then cells were activated with 10 μg/mL LPS dissolved in growth media for 4 h. Cells lysates were obtained, TF expression and activity were measured with western blot analysis and tissue factor human chromogenic activity assay kit respectively. Immunocytochemical fluorescence confocal microscopy analysis after amaranth peptides treatment was performed. The expression profile of thrombosis-inflammation related proteins was performed with a Human XL Cytokine Array.Results: The AF was found to inhibit TF expression (IC50= 0.39mg/mL) and activity (IC50 = 0.58 mg/mL). Localization of TF showed that treated monocytes decreased 49.01 % in membrane and increased two fold in nuclei compared to positive control, indicating that the treatment reduces active surface TF and decreases the initiation of the coagulation cascade. Moreover, cytokines array indicated that AF peptides produced suppression in the expression of MIP-3α, Interleukin-1β, Interleukin-1 α, TARC, PDGF-AA and Pentaxin 3 compared to Ctrl+, with reductions of 78.8%, 61.8%, 54.1%, 42.6%, 37.8% and 37.9% respectively. According to these results the cytokines with the highest inhibition of expression are involved in the induction of NF-κB pathway and platelet adhesion, aggregation, activation process.Conclusions: Results provide potential mechanistic information on the antithrombotic and anti-inflammatory effect of AF, indicating that amaranth peptides could inhibit TF expression by producing a negative feedback regulator role over NF-κB pathway. This effect could result in the prevention of hypercoagulability states and subsequent thrombosis and inflammation pathologies.