Development of novel agents using the method of modulating the transcriptional activity of apo (a) gene

利用调节apo（a）基因转录活性的方法开发新药物

• 批准号: 10557105
• 负责人: MATSUMOTO Toshio
• 金额: $ 8.32万
• 依托单位: University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557105/
• 关键词: lipoprotein (a); apolipoprotein (a); atherosclerosis; aspirin; transcriptional factor; hepatocyte; endothelial cell; 血管内及細胞; IL-6; 転写活性

Increased lipoprotein (Lp)(a) level is an independent risk factor for atheroscrelotic diseases and is observed in a frequency of 10〜15% in Japanese population. At present, there is few clinically effective agents with less adverse effects. We have developed a novel assay system in which a 1.4 kb in length of promoter region of apolipoprotein (apo)(a) is ligated in 5' end of luciferase plasmid. Among several agents we examined, we found that aspirin reduced luciferase activity in HepG2 cells and also suppressed mRNA expression of apo (a) in human hepatocytes. In vitro examination showed that aspirin decreased apo (a) production from hepatocytes by 30%. Based on these findings, we analyzed the effect of aspirin on serum Lp (a) levels in patients with atherosclerotic diseases and found that aspirin reduced Lp (a) levels by 20% in patients with high Lp (a) levels (more than 30 mg/dl) but not with normal Lp (a) levels (less than 30 mg/dl). Furthermore, the values of decrease in serum Lp (a) by aspirin in patients with high Lp (a) were positively correlated with the baseline Lp (a) levels. The difference of serum Lp (a) levels in patients containing same apo (a) isoform sizes is considered to be due to the difference of the transcriptional activity of apo (a) gene. Therefore, aspirin lowers serum Lp (a) levels in patients with high Lp (a) potentially via the reduction of apo (a) gene transcription.We next aimed to reveal the novel action of aspirin on the reduction of high Lp (a) levels via an independent mechanism from arachidonate pathway. Using enhanced subtractional hybridization analysis, we found a new nuclear factor with unknown function and are examining the effect of activation on the expression levels of apo (a).

脂蛋白（Lp）（a）水平升高是动脉粥样硬化性疾病的独立风险因素，在日本人群中观察到的频率为10 - 15%。目前，临床上有效的药物很少，不良反应较少。我们将载脂蛋白（apo）（a）的启动子区连接到荧光素酶质粒的5'端，建立了一种新的检测系统。在我们检测的几种药物中，我们发现阿司匹林降低了HepG 2细胞中的荧光素酶活性，也抑制了人肝细胞中apo（a）的mRNA表达。体外实验表明阿司匹林使肝细胞apo（a）的生成减少了30%。基于这些发现，我们分析了阿司匹林对动脉粥样硬化性疾病患者血清Lp（a）水平的影响，发现阿司匹林可使高Lp（a）水平（超过30 mg/dl）但Lp（a）水平正常（低于30 mg/dl）的患者的Lp（a）水平降低20%。此外，高脂蛋白（a）患者阿司匹林降低血清脂蛋白（a）的值与基线脂蛋白（a）水平正相关。载脂蛋白（a）亚型大小相同的患者血清Lp（a）水平的差异可能是由于载脂蛋白（a）基因转录活性的差异。因此，阿司匹林可能通过降低载脂蛋白（a）基因的转录水平来降低高脂蛋白（a）患者的血清Lp（a）水平。利用增强的消减杂交分析，我们发现了一种新的功能未知的核因子，并正在研究激活对载脂蛋白（a）表达水平的影响。

项目成果

Kagawa A. et al.: "Aspirin reduces apolipoprotein(a)(apo(a)) production in human hepatocytes by suppression of apo(a)gene transcription"J.Biol.Chem.. 274. 34111-34115 (1999)

Kakawa A.等人：“阿司匹林通过抑制apo(a)基因转录来减少人肝细胞中载脂蛋白(a)(apo(a))的产生”J.Biol.Chem.. 274. 34111-34115 (1999)

Watanabe A.et al.: "Analysis of the effect of aspirin on the transcriptional activity of apo (a) gene"Proceedings of The Japan Society of Clinical Biochemistry and Metabolism. 35. 37 (1998)

Watanabe A.等人：“阿司匹林对apo（a）基因转录活性的影响分析”日本临床生物化学和代谢学会会刊。

Azuma H.et al: "An in vitro system for identifying agents capable of changing serum lipoprotein(a) concentration by regulating the transcriptional activity of the apolipoprotein(n) gene promoter" Biochem.Biophys.Res.Commun.227. 570-575 (1996)

Azuma H.等人：“一种体外系统，用于鉴定能够通过调节载脂蛋白（n）基因启动子的转录活性来改变血清脂蛋白（a）浓度的试剂”Biochem.Biophys.Res.Commun.227。

赤池雅史 他: "高リポ蛋白(a)血症に及ぼすアスピリンの影響"四国医学雑誌. 55. 109-114 (1999)

Masashi Akaike 等：“阿司匹林对高脂蛋白（a）血症的影响”四国医学杂志 55. 109-114（1999）。

Kagawa A.et al: "Aspirin reduces apolipoprotein(a) (apo(a)) production in human hepato cytos by suppression of apo(a) gene transcription"J.Biol.Chem.. 274・48. 34111-34115 (1999)

Kakawa A.等人：“阿司匹林通过抑制apo(a)转录基因来减少人肝细胞中载脂蛋白(a) (apo(a))的产生” J.Biol.Chem.. 274・48 (1999) ）