Gene therapy for atherosclerosis by inhibition of nuclear transcriptional factor

通过抑制核转录因子进行动脉粥样硬化的基因治疗

基本信息

批准号：
07457174
负责人：
KITAJIMA Isao
金额：
$ 1.02万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

The proliferation of vascular smooth muscle cells (VSMCs) is the most crucial cause of artherosclerosis. High expression of thrombin receptor (TR) in athrosclerotic lesions indicates a possible role for athrogenesis. In the present study, we demonstrated that post TR signaling were mediated by several protein kinases resposible for nuclear transcriptional factor, NF-kappaB activation and thrombin-inducible genes had the kappaB sequence in the regulatory elements. Thrombin stimulation in VSMCs resulted in a biphasic activation of NF-kappaB,the early phase of which indicated nuclear translocation of NF-kappaB and the late phase of which required new synthesis, resulting in proliferation of VSMC.Furthermore, treatment of VSMCs with antisense p65 oligodeoxynucleotides (ODNs) of NF-kappaB inhibited thrombin-stimulated growth of VSMCs in vitro. Next, we examined whether the antisense ODNs for NF-kappaB could affect in vivo. We used the mouse model of lipopolysaccharide (LPS) induced septic shock, because it is associated with activation of NF-kappaB.We showed that all mice treated with LPS ultimately died within 48 hrs, while mice pretreated with the antisense NF-kappaB ODNs had a significantly better outcome and the survival rate was 60%. This suggested the greatest advantage antisense NF-kappaB provides an important new therapeutic approach for the treatment with athrerosclerosis and LPS-induced septic shock in vitro and in vivo. Moreover, we have studied a novel biomaterial vessle using aramid-silicon to intruduce the antisense ODNs.

血管平滑肌细胞（VSMC）的增殖是动脉粥样硬化的最主要原因。动脉粥样硬化病变中凝血酶受体（TR）的高表达表明其可能在动脉粥样硬化形成中发挥作用。在本研究中，我们证明TR后信号传导是由几种负责核转录因子、NF-kappaB激活的蛋白激酶介导的，并且凝血酶诱导基因在调节元件中具有kappaB序列。 VSMCs中凝血酶刺激导致NF-kappaB双相激活，早期表明NF-kappaB核转位，后期需要新的合成，导致VSMC增殖。此外，反义p65处理VSMCs NF-kappaB 的寡脱氧核苷酸 (ODN) 在体外抑制凝血酶刺激的 VSMC 生长。接下来，我们检查了 NF-kappaB 的反义 ODN 是否会在体内产生影响。我们使用脂多糖（LPS）诱导感染性休克的小鼠模型，因为它与 NF-kappaB 的激活有关。我们发现所有用 LPS 处理的小鼠最终在 48 小时内死亡，而用反义 NF-kappaB ODN 预处理的小鼠则死亡。结果明显更好，存活率为 60%。这表明反义NF-κB的最大优势为体外和体内治疗动脉粥样硬化和LPS诱导的感染性休克提供了一种重要的新治疗方法。此外，我们还研究了一种使用芳纶硅引入反义ODN的新型生物材料容器。