A novel gene regulator, pyrrole–imidazole polyamide targeting ABCA1 gene increases cholesterol efflux from macrophages and plasma HDL concentration
A novel gene regulator, pyrrole–imidazole polyamide targeting ABCA1 gene increases cholesterol efflux from macrophages and plasma HDL concentration
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DOI：
10.1007/s00109-013-1118-x
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发表时间：
2014-01
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通讯作者：
A. Tsunemi;T. Ueno;N. Fukuda;Takayoshi Watanabe;K. Tahira;Akira Haketa;Yoshinari Hatanaka;Sho Tanaka;Taro Matsumoto;Y. Matsumoto;H. Nagase;M. Soma
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作者：
A. Tsunemi;T. Ueno;N. Fukuda;Takayoshi Watanabe;K. Tahira;Akira Haketa;Yoshinari Hatanaka;Sho Tanaka;Taro Matsumoto;Y. Matsumoto;H. Nagase;M. Soma
吡咯-咪唑（PI）聚酰胺是一种新型的抗核酸酶化合物，通过与DNA小沟结合来抑制转录因子。设计并评价了靶向小鼠ABCA 1并增加ABCA 1基因表达的PI聚酰胺作为增加血浆HDL浓度的试剂。设计PI聚酰胺以结合小鼠ABCA 1启动子的激活蛋白-2结合位点。使用RAW 264细胞通过实时RT-PCR和蛋白质印迹法评价该PI聚酰胺对ABCA 1表达的影响。在C57 B6小鼠中检测了该聚酰胺对ABCA 1基因表达和血浆HDL水平的体内作用。每2天通过尾静脉注射1毫克/千克体重的PI聚酰胺，持续1周，并评价血浆脂质谱。凝胶迁移率变动分析显示PI聚酰胺与靶DNA具有特异性结合。用1.0 μM PI聚酰胺处理RAW 264细胞显著增加ABCA 1 mRNA表达。PI聚酰胺也显着增加载脂蛋白AI介导的HDL在RAW 264细胞的生物合成。PI聚酰胺处理显著增加了载脂蛋白AI介导的细胞胆固醇流出。PI聚酰胺显著增加C57 B6小鼠肝脏中ABCA 1 mRNA的表达。PI聚酰胺给药可增加血浆HDL浓度。所有HDL亚组分在PI聚酰胺给药后显示出增加的趋势。设计的靶向ABCA 1的PI聚酰胺成功地增加了ABCA 1表达和HDL生物合成。一种新型的吡咯-咪唑（PI）聚酰胺与ABCA 1结合，PI聚酰胺干扰AP-2 β蛋白与ABCA 1基因启动子的结合，PI聚酰胺抑制AP-2 β蛋白与ABCA 1基因启动子的结合，PI聚酰胺抑制AP-2 β蛋白与ABCA 1基因启动子的结合。PI聚酰胺可增加ABCA 1蛋白和载脂蛋白AI介导的HDL生物合成。PI聚酰胺是一种高分子材料，用于预防动脉粥样硬化疾病的新基因调节剂。
AbstractPyrrole–imidazole (PI) polyamides are nuclease-resistant novel compounds that inhibit transcription factors by binding to the minor groove of DNA. A PI polyamide that targets mouse ABCA1 and increases ABCA1 gene expression was designed and evaluated as an agent to increase plasma HDL concentration. A PI polyamide was designed to bind the activator protein-2 binding site of the mouse ABCA1 promoter. The effect of this PI polyamide on ABCA1 expression was evaluated by real-time RT-PCR and Western blotting using RAW264 cells. In vivo effects of this polyamide on ABCA1 gene expression and plasma HDL level were examined in C57B6 mice. One milligram per kilogram of body weight of PI polyamide was injected via the tail veins every 2 days for 1 week, and plasma lipid profiles were evaluated. PI polyamide showed a specific binding to the target DNA in gel mobility shift assay. Treatment of RAW264 cells with 1.0 μM PI polyamide significantly increased ABCA1 mRNA expression. PI polyamide also significantly increased apolipoprotein AI-mediated HDL biogenesis in RAW264 cells. Cellular cholesterol efflux mediated by apolipoprotein AI was significantly increased by the PI polyamide treatment. PI polyamide significantly increased expression of ABCA1 mRNA in the liver of C57B6 mice. Plasma HDL concentration was increased by PI polyamide administration. All of the HDL sub-fractions showed a tendency to increase after PI polyamide administration. The designed PI polyamide that targeted ABCA1 successfully increased ABCA1 expression and HDL biogenesis. This novel gene-regulating agent is promising as a useful compound to increase plasma HDL concentration.Key messagesA novel pyrrole–imidazole (PI) polyamide binds to ABCA1.PI polyamide interfered with binding of AP-2ɑ protein to the ABCA1 gene promoter.PI polyamide inhibited the AP-2ɑ-mediated reduction of ABCA1 gene and protein expression.PI polyamide increased ABCA1 protein and apolipoprotein AI mediated HDL biogenesis.PI polyamide is a new gene regulator for the prevention of atherosclerotic diseases.