Arterial hypertension modeled by in vitro treatment of red blood cells with trypsin

Rheological studies of erythrocyte membrane (RBC) are useful to analyze and model the microcirculatory disturbances observed in arterial hypertension (HT). Previous studies have shown that erythrocyte anionic charge (EAC) decreases affecting intercellular interactions. To understand RBC disturbances in HT, modeling in vitro by the trypsin action in normotensive donor samples and comparative analysis were performed. Measurements of EAC by spectrophotometric analysis with the Alcian Blue dye were carried out to quantify the effect of trypsin on RBC. Erythrocyte aggregation kinetics was analyzed with an Erythroaggregameter determining the time required to reach 50% of maximum RBC aggregation (t50%). Results show a decrease in the EAC in erythrocytes of normotensive donors enzymatically treated (p<0.05) and hypertensive patients (p<0.005) compared to control. Similar EAC values of erythrocytes from HT and RBC treated with trypsin solution 2mg/mL were found. There was a significant decrease (p<0.05) in t50% with increasing trypsin concentration, yielding similar t50% values for RBCs of HT with healthy RBCs treated with trypsin solution 2mg/mL. In vitro modeling proved to reproduce the RBC alterations in HT, establishing that 2mg/mL trypsin treatment produces similar changes to those found in this pathology.