CAJ | 학술논문

In order to study whether the endothelial cells (ECs) with lipid peroxidation induced by diamide can express and secrete macrophage inflammatory protein 1α (MIP-1α), the expression ofMIP-1α protein in the cells was detected by cell enzyme-linked immunosorbent assay (EL ISA) andthat of MIP-1α mRNA was determined by cell in situ hybridization and nuclease S1 protection assayafter the ECs were exposed to different concentrations of diamide for 4 h. The chemotactic activityof MIP-1α was tested by micropore filter method using modified Boyden chambers. Cell ELISAshowed that the expression of MIP-1α protein in endothelial cells exposed to 1 μmol/L, 5 μmol/Land 10 μmol/L diamide was 1.9-fold, 2.3-fold and 1.7-fold respectively as much as that in the con-trol cells, which was statistically significant by analysis of variance. In situ hybridization revealedthat the mRNA expression of ECs treated with 1 μmol/L, 5 μmol/L and 10 μmol/L diamide was1.3-fold, 3.0-fold and 1.7-fold as much as that in the control group, which had statistical signifi-cance (F=188.93, P＜0.01). The mRNA expression in 5 μmol/L dimide treated ECs, measuredby nuclease S1 protection assay, was 3.4-fold as much as that in the control group(t=8.70, P＜0.05). Chemotactic response(99.50±4.31 μm) to the culture medium conditioned by 5 μmol/L di-amide treated ECs , which was stronger than that(66.47±3.25 μm) conditioned by the ECs (F=404.31, P＜0.05), was significantly decreased (F=192.25, P＜0.05) after adding MIP-1α anti-body. It suggests that diamide, a lipid peroxidation inducer, could stimulate ECs to produce highlevel of MIP-1α, and might play an important role in atherogenesis by promoting the migration ofperipheral blood monocytes into arterial intima.