动脉粥样硬化细胞模型建立及细胞内铁代谢紊乱初步研究

发布时间：2019-01-08 16:28

【摘要】：目的建立动脉粥样硬化(AS)细胞模型,研究铁代谢与AS关系及AS斑块中巨噬细胞铁代谢紊乱的分子机制,为临床干预铁代谢及防治AS提供理论依据。方法采用RAW264.7细胞,以氧化型低密度脂蛋白(ox-LDL)诱导制备泡沫细胞,油红O染色并通过酶联免疫法检测胞内脂质(总胆固醇、游离胆固醇)验证AS泡沫细胞模型建立。免疫印迹法检测铁代谢相关铁蛋白(FT)、膜铁转运蛋白(FPN)1,免疫组化、免疫荧光法检测和定位巨噬细胞中FPN1表达。结果 ox-LDL诱导制备的细胞质内有大量红色脂质颗粒,较多脂质融合成大油滴状,符合泡沫细胞形态特征;泡沫细胞内有大量脂质堆积,胆固醇酯(CE)占比明显高于正常细胞(P0.05);与正常对照组相比,泡沫细胞内FT含量明显高于正常细胞,FPN1含量增加且主要存在于细胞质内。结论泡沫细胞中铁代谢紊乱,细胞内大量聚集。FPN1非细胞膜定位阻止铁从巨噬细胞中有效排出并加重泡沫细胞中铁累积,可能加剧AS斑块形成和发展。
[Abstract]:Objective to establish a model of atherosclerotic (AS) cells and to study the relationship between iron metabolism and AS and the molecular mechanism of iron metabolism disorder in macrophages in AS plaques so as to provide a theoretical basis for clinical intervention of iron metabolism and prevention and treatment of AS. Methods RAW264.7 cells were induced by oxidized low density lipoprotein (ox-LDL) to prepare foam cells. Oil red O staining was used to detect intracellular lipid (total cholesterol, free cholesterol) of AS foam cells. The expression of iron metabolism-related ferritin (FT), membrane iron transporter (FPN) 1) in macrophages was detected by immunoblotting and immunohistochemical staining. The expression of FPN1 in macrophages was detected by immunofluorescence assay. Results there were a large number of red lipid particles in the cytoplasm induced by ox-LDL, and more lipids were fused into large oil droplets, which accorded with the morphological characteristics of foam cells. There was a large amount of lipid accumulation in foam cells, and the proportion of cholesterol ester (CE) was significantly higher than that in normal cells (P0.05). Compared with the control group, the content of FT in foam cells was significantly higher than that in normal cells, and the content of FPN1 was increased and mainly existed in the cytoplasm. Conclusion the iron metabolism in foam cells is disordered and a large amount of intracellular concentration is accumulated. The localization of FPN1 non-cell membrane prevents iron from effectively excreting from macrophages and exacerbates iron accumulation in foam cells, which may aggravate the formation and development of AS plaques.
【作者单位】：江苏大学附属武进医院介入血管科;南京大学医学院附属鼓楼医院血管外科;
【基金】：国家自然科学基金(81370387)
【分类号】：R543.5

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