Author + information
To culture circulating endothelial progenitor cells at coronary atherosclerosis, evaluate eNOS levels in this culture.
The 5 ml blood was drawn from the peripheral vein of 12 man patients (stable angina pectoris 4, acute myocardial infarction 4, healthy people 4). Mononuclear cells were isolated with the use of a Ficoll density gradient SepMate15 (Stem Cell Technologies Inc, Canada) according to standard protocols. In an endothelial basal medium “Using CFU-Hill Liquid Medium Kit” (Stem Cell Technologies Inc, Canada) with supplements, mononuclear cells were seeded on human fibronectin-coated plates (Stem Cell Technologies Inc, Canada). After 6 days in vitro, two independent investigators counted endothelial colony-forming units after giemsa staining. eNOS enzyme titers were determined by ELISA according to the manufacturer's recommended protocol (Human eNOS, R and D Systems Inc. USA) in the cells culture.
The people were 52±7.3 years (range, 41 to 62). The number of EPC colonies had direct significantly correlation (r=0.621, p=0.000) with the eNOS enzyme levels of this culture. Furthermore, The number of EPC colonies increases with accordance of patients with stable angina pectoris, acute myocardial infarction, healthy people with the statistical significance (F=17.3, p=0.000): stable angina (2.6±0.47 colony/well), acute myocardial infarction (6.7±0.81 colony/well), healthy people (10.5±1.34 colony/well).
One way ANOVA test of eNOS enzyme levels in patients with stable angina pectoris (5.2±0.61pg/ml), acute myocardial infarction (8.7±1.49pg/ml) and healthy people (13.7±2.48pg/ml). the significant difference (F=6.2, p=0.003) was observed among the three groups.
Number of EPC colonies (F=17.3, p=0.000) and eNOS enzyme levels (F=6.2, p=0.003) decrease at stable angina pectoris, indicate more than endothelial dysfunction.