Mipu1 overexpression protects macrophages from oxLDL-induced foam cell formation and cell apoptosis.
Mipu1 overexpression protects macrophages from oxLDL-induced foam cell formation and cell apoptosis.
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DOI：
10.1089/dna.2014.2501
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发表时间：
2014-12
影响因子：
3.1
通讯作者：
Shunlin Qu;W. Fan;Chi Zhang;Fang Guo;Dan Han;W. Pan;Wei Li;D. Feng;Zhi-Sheng Jiang
中科院分区：
文献类型：
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作者：
Shunlin Qu;W. Fan;Chi Zhang;Fang Guo;Dan Han;W. Pan;Wei Li;D. Feng;Zhi-Sheng Jiang
心肌缺血预处理上调蛋白1（myocardial ischemic preconditioning upregulated protein 1，Mipu 1）是一个新的N-末端Kruppel相关盒（KRAB）/C2 H2锌指超家族蛋白，对氧化应激诱导的H9 c2细胞凋亡具有保护作用。本研究旨在探讨Mipu 1过表达对氧化低密度脂蛋白（oxLDL）诱导的泡沫细胞形成和细胞凋亡的影响及其可能机制。采用健康新西兰大白兔建立动脉粥样硬化模型，采用全自动生化分析仪检测血清甘油三酯、总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇水平。苏丹IV染色用于检测动脉粥样硬化病变。以RAW264.7巨噬细胞系为实验材料。油红O染色法、高效液相色谱法和Dil标记脂蛋白法分别定性和定量检测胆固醇蓄积。流式细胞术检测细胞凋亡。实时定量聚合酶链反应（PCR）检测与胆固醇转运相关的主要蛋白ABCA 1、ABCG 1、SR-BI和CD 36的mRNA表达。Western blot检测Mipu 1蛋白的表达。高脂饮食组苏丹IV染色可见动脉粥样硬化病变。高脂饮食组在第10周时Mipu 1的表达显著低于标准饮食组，而CD 36的表达显著高于标准饮食组。Mipu 1过表达降低oxLDL诱导的胆固醇蓄积、oxLDL摄取、细胞凋亡和切割的caspase-3。Mipu 1过表达抑制oxLDL诱导的CD 36 mRNA和蛋白表达，但对ABCA 1、ABCG 1和SR-BI的mRNA表达无明显抑制作用。Mipu 1过表达抑制oxLDL诱导的泡沫细胞形成和细胞凋亡Mipu 1过表达减少巨噬细胞的脂质摄入，可能与oxLDL存在下CD 36表达下调有关。
Mipu1 (myocardial ischemic preconditioning upregulated protein 1) is a novel N-terminal Kruppel-associated box (KRAB)/C2H2 zinc finger superfamily protein, that displays a powerful effect in protecting H9c2 cells from oxidative stress-induced cell apoptosis. The present study aims to investigate the effect of Mipu1 overexpression on oxidized low-density lipoprotein (oxLDL)-induced foam cell formation, cell apoptosis, and its possible mechanisms. New Zealand healthy rabbits were used to establish atherosclerosis model, and serum levels of triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were detected by an automatic biochemical analyzer. Sudan IV staining was used to detect atherosclerotic lesions. The RAW264.7 macrophage cell line was selected as the experimental material. Oil red O staining, high-performance liquid chromatography, and Dil-labeled lipoprotein were used to detect cholesterol accumulation qualitatively and quantitatively, respectively. Flow cytometry was used to determine cell apoptosis. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression of the main proteins that are associated with the transport of cholesterol, such as ABCA1, ABCG1, SR-BI, and CD36. Western blot analysis was used to detect the protein expression of Mipu1. There were atherosclerotic lesions in the high-fat diet group with Sudan IV staining. High-fat diet decreased Mipu1 expression and increased CD36 expression significantly at the 10th week compared with standard-diet rabbits. Mipu1 overexpression decreased oxLDL-induced cholesterol accumulation, oxLDL uptake, cell apoptosis, and cleaved caspase-3. Mipu1 overexpression inhibited the oxLDL-induced CD36 mRNA and protein expression, but it did not significantly inhibit the mRNA expression of ABCA1, ABCG1, and SR-BI. Mipu1 overexpression inhibits oxLDL-induced foam cell formation and cell apoptosis. Mipu1 overexpression reduces the lipid intake of macrophages and might be associated with the downregulation of CD36 expression in the presence of oxLDL.