论文部分内容阅读
为探讨Daxx对氧化型低密度脂蛋白(oxidized low-density lipoprotein,Ox-LDL)诱导巨噬细胞胆固醇蓄积和凋亡的介导作用及其可能的分子机制,用高效液相色谱法检测细胞内胆固醇含量,油红O染色观察细胞内脂滴的形成情况,流式细胞术和吖啶橙/溴化乙锭(AO/EB)染色法研究Ox-LDL对细胞凋亡的影响,Realtime RT-PCR检测细胞内Daxx mRNA的表达水平,Western blot检测caveolin-1蛋白的表达,用特异性siRNA沉默Daxx在RAW264.7细胞中的表达.Ox-LDL上调Daxx mRNA和caveolin-1的表达、增加细胞内胆固醇含量、促使RAW264.7细胞凋亡,用特异性siRNA干扰Daxx在RAW264.7细胞中的表达能降低caveolin-1的表达、减少细胞内胆固醇含量、以及抑制细胞凋亡.上述结果表明,Daxx对Ox-LDL诱导RAW264.7巨噬细胞胆固醇蓄积和凋亡具有介导作用,这一作用可能与Daxx上调caveolin-1的表达有关。
To investigate the effect of Daxx on macrophage cholesterol accumulation and apoptosis induced by oxidized low-density lipoprotein (Ox-LDL) and its possible molecular mechanism, we used high performance liquid chromatography The effect of Ox-LDL on apoptosis was investigated by flow cytometry and AO / EB staining. Realtime RT- PCR was used to detect the expression of Daxx mRNA in the cells.Western blot was used to detect the expression of caveolin-1 protein and silencing the expression of Daxx in RAW264.7 cells with specific siRNA.Ox-LDL upregulated the expression of Daxx mRNA and caveolin-1, The content of cholesterol in RAW264.7 cells induced by apoptosis and the expression of caveolin-1 in RAW264.7 cells by Doxx with specific siRNA could reduce the intracellular cholesterol and inhibit the apoptosis of RAW264.7 cells.The above results showed that, Daxx can mediate Ox-LDL-induced cholesterol accumulation and apoptosis in RAW264.7 macrophages, which may be related to upregulation of caveolin-1 expression by Daxx.