花色苷对小鼠腹腔巨噬泡沫细胞胆固醇外流的影响及其分子机制探讨

The effect of anthocyanins on cholesterol efflux from mouse peritoneal macrophage-derived foam cells and its possible molecular mechanism

目的研究花色苷对小鼠腹腔巨噬泡沫细胞胆固醇外流的影响,探讨花色苷抗动脉粥样硬化的分子机制。方法以50μg/ml 氧化型低密度脂蛋白培养小鼠腹腔巨噬细胞24 h 形成巨噬泡沫细胞。用1、10、100μmoL/L 矢车菊定-3-葡萄糖苷(Cy-3-g)和芍药定-3-葡萄糖苷(Pn-3-g)分别培养巨噬泡沫细胞,酶学荧光法检测培养液中胆固醇含量,用定量 PCR 和激光共聚焦显微镜检测三磷酸腺苷结合盒转运体 A1(ABCA1)的表达。结果花色苷 Cy-3-g 和 Pn-3-g 能够促进巨噬泡沫细胞胆固醇外流,这一作用存在时间和剂量-依赖关系。花色苷还能促进巨噬泡沫细胞 ABCA1 mRNA 和蛋白的表达,ABCA1抑制剂4,4’-二异硫氰酸二丙乙烯-2,2'-二磺酸能够阻断花色苷诱导巨噬泡沫细胞胆固醇外流。结论花色苷 Cy-3-g 和 Pn-3-g 通过调控 ABCA1的表达从而促进巨噬泡沫细胞胆固醇外流。

Objective To investigate the potential role of anthocyanins on modulating cholesterol efflux in mouse peritoneal macrophage-derived foam cells and related molecular mechanisms. Methods The macrophages were isolated from pathogen-free NIH mice and were loaded with 50 μg/ml oxLDL for 24 hours, newly formed foam cells were then treated with anthocyanins （cyanidin-3-glucoside, Cy-3-g ; or peonidin-3- glucoside,Pn-3-g） at the concentrations of 1 μmol/L, 10 μmol/L, 100 μmol/L for 0 to 36 hours, respectively. The enzymatic-fluorescent method was used to determine cholesterol content in culture medium. ABCA1 expressions at mRNA and protein level were detected by real-time PCR and confocal microscope. Results Cholesterol efflux of macrophage-derived foam cells increased in a time- and dose-dependent manner post anthocyanins treatment. ABCA1 expressions at mRNA and protein levels were also significantly enhanced after anthocyanins treatment in these cells and these effects could be blocked by co-treatment with DIDS, an inhibitor of the transport activities of ABCA1 and blocker of apoAI-mediated cholesterol efflux. Conclusion These data demonstrate that anthocyanins induce cholesterol efflux from mouse peritoneal macrophage-derived foam cells via regulating ABCA1 expression.