Platelet Aggregation Increased by Advanced Glycated Hemoglobin

Abstract

Introduction: Many studies have provided strong evidence for an association between diabetes complications and an increase in platelet (PLT) reactivity. Though some metabolic abnormalities have been reported as the major causes of this reactivity and malfunction, the precise mechanism has not been fully elucidated. Aim: The aim of this study is to investigate the effect of advanced glycation end product of hemoglubin (AGE-Hb) on human PLT reactivity and malfunction. Materials and Methods: A solution of bovine hemoglobin was prepared with fructose in phosphate buffer. The solution was then sterilized and incubated under sterile conditions at 37°C in the dark. The control solution was prepared in the same way, but without fructose. Human PLTs were isolated and prepared from 15 healthy volunteers, men and women. Blood was collected in the morning from fasting healthy subjects using a 21-gauge needle with 117 mM sodium citrate (1:9 v/v), as an anticoagulant. Fluorescence measurements were performed using a Hitachi F-4500 spectrofluorometer. PLT aggregation was measured using the photometric system Packs-4 aggregometer. Results: The relative fluorescence intensity increased in Hb samples but not in controls that incubated with fructose. PLTS aggregation did not change in controls while after incubation with 10, 22, 30-days fructose-glycated Hb decreased by 10, 12, 30%, respectively. Taken together our data show that PLT secondary phase of ADP-induced aggregation gradually affected with advanced increase in Hb glycation.