目的 观察氧化低密度脂蛋白（ox-LDL）诱导巨噬细胞泡沫化后血管紧张素转化酶2（ACE2）的表达变化与氧化应激损伤的关系，探讨ACE2的抗氧化作用。 方法 用不同浓度ox-LDL诱导小鼠巨噬细胞RAW264.7泡沫化，油红O染色观察巨噬细胞内脂质堆积变化，并定量分析细胞内脂滴含量，通过检测细胞上清中丙二醛（MDA）含量鉴定细胞氧化损伤程度，实时荧光定量PCR检测ACE2及MAS mRNA表达，化学比色法检测超氧化物歧化酶（SOD）活性与还原型谷胱甘肽（GSH）的含量。 结果 不同浓度(20、40、60、80 μg·ml-1) ox-LDL处理巨噬细胞24 h，油红O染色可见巨噬细胞变大、变圆，胞浆内脂滴含量呈剂量依赖性增加；用60 μg·ml -1ox-LDL诱导巨噬细胞泡沫化模型组中MDA含量较空白组明显升高（P<0.01)；Real-Time PCR结果显示，模型组ACE2和MAS mRNA 表达水平较空白组显著减少(P<0.05)；与空白组比较，模型组中SOD活性与GSH 含量显著降低(P<0.05或P<0.01)。 结论 巨噬细胞泡沫化氧化损伤进程与ACE2及其下游受体MAS表达下调密切相关，提示ACE2具有一定的抗氧化作用，其机制可能与SOD及GSH有关。
Abstract
Objective To investigate the change of ACE2 expression in the macrophage foam cell induced by ox-LDL. Methods Different concentrations of ox-LDL were used to induce macrophage cell RAW264.7 into foam cells, the intracellular lipid droplets amounts were observed by oil red O staining, the content of MDA was measured to identify oxidative damage, Real-Time PCR was used to detect ACE2 and MAS mRNA expression, the activity of SOD and the content of GSH were measured by chemocolorimetry. Results The macrophages became larger and rounder after treated by ox-LDL (20、40、60、80μg·ml-1) for 24h by oil red O staining, the amount of lipid droplets in the cytoplasm increased in a dose-dependent manner. The MDA content was increased significantly compare with the control group using 60μg·ml-1 ox-LDL treatment (P<0.01), Real-time PCR results showed that ACE2 and MAS mRNA expression in model group were decreased (P<0.05), and the SOD activity and the GSH content were also decreased (P<0.01 or P<0.05) significantly in model group. Conclusion Macrophage foam cell formation is closely related with the down-regulation of ACE2 and MAS expression, and that means ACE2 has anti-oxidative effect which may be related to SOD and GSH.
关键词
氧化低密度脂蛋白 /
巨噬细胞 /
泡沫细胞 /
ACE2 /
抗氧化作用
Key words
ox-LDL /
Macrophage /
Foam cell /
ACE2 /
Anti-oxidative effect
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