【摘要】：目的建立動脈粥樣硬化(AS)細(xì)胞模型,研究鐵代謝與AS關(guān)系及AS斑塊中巨噬細(xì)胞鐵代謝紊亂的分子機(jī)制,為臨床干預(yù)鐵代謝及防治AS提供理論依據(jù)。方法采用RAW264.7細(xì)胞,以氧化型低密度脂蛋白(ox-LDL)誘導(dǎo)制備泡沫細(xì)胞,油紅O染色并通過酶聯(lián)免疫法檢測胞內(nèi)脂質(zhì)(總膽固醇、游離膽固醇)驗證AS泡沫細(xì)胞模型建立。免疫印跡法檢測鐵代謝相關(guān)鐵蛋白(FT)、膜鐵轉(zhuǎn)運(yùn)蛋白(FPN)1,免疫組化、免疫熒光法檢測和定位巨噬細(xì)胞中FPN1表達(dá)。結(jié)果 ox-LDL誘導(dǎo)制備的細(xì)胞質(zhì)內(nèi)有大量紅色脂質(zhì)顆粒,較多脂質(zhì)融合成大油滴狀,符合泡沫細(xì)胞形態(tài)特征;泡沫細(xì)胞內(nèi)有大量脂質(zhì)堆積,膽固醇酯(CE)占比明顯高于正常細(xì)胞(P0.05);與正常對照組相比,泡沫細(xì)胞內(nèi)FT含量明顯高于正常細(xì)胞,FPN1含量增加且主要存在于細(xì)胞質(zhì)內(nèi)。結(jié)論泡沫細(xì)胞中鐵代謝紊亂,細(xì)胞內(nèi)大量聚集。FPN1非細(xì)胞膜定位阻止鐵從巨噬細(xì)胞中有效排出并加重泡沫細(xì)胞中鐵累積,可能加劇AS斑塊形成和發(fā)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.
【作者單位】： 江蘇大學(xué)附屬武進(jìn)醫(yī)院介入血管科;南京大學(xué)醫(yī)學(xué)院附屬鼓樓醫(yī)院血管外科;
【基金】：國家自然科學(xué)基金(81370387)
【分類號】：R543.5

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