The establishment of atherosclerotic cell model and the preliminary study of intracellular iron metabolism disorder / 介入放射学杂志

ABSTRACT

Objective To establish the atherosclerosis (AS) cell model,to study the relationship between the iron metabolism and AS,and to explore the molecular mechanisms of iron metabolism disorders within the macrophages in AS plaques so as to provide theoretical basis for clinical intervention of iron metabolism and for the prevention and treatment of AS.Methods RAW264.3 cells were used to prepare foam cells through oxidized low density lipoprotein (ox-LDL) induced oxidation.In order to verify the foam cells,oil red staining and enzyme-linked immunosorbent were employed to assay the total lipids and free lipids (including total cholesterol and free cholesterol) in the foam cells.Western blotting was adopted to test the levels of iron metabolism-associated ferritin (FT) and ferroportin1 (FPN1);immunohistochemistry and immunofluorescence methods were used to determine and locate the expressions of FPN1 in foam cells.Results Large amounts of red lipid granules could be seen within the cytoplasm of ox-LDL-induced foam cells,and a great quantity of lipid fused into the form of large oil droplets,which was consistent with the morphological characteristics of foam cells.Massive lipid was accumulated in the foam cells,and the proportion of cholesterol ester (CE) in the foam cells was significantly higher than that in the normal cells (P＜ 0.05).Compared with the normal control group,FT content in the foam cells was obviously higher than that in the normal cells;the content of FPN1 was increased,and the increased content of FPN1 mainly existed in cytoplasm.Conclusion In the foam cells the iron metabolism is disordered and a great quantity of FPN1 is accumulated in cytoplasm.The non-cell-membrane localization of FPN1 can prevent iron from being effectively discharged from macrophages and can increase the accumulation of iron in foam cells,which may aggravate the formation and development of AS plaques.