Study of the molecular mechanism and clinical implication of a novel apoptosis gene

新型凋亡基因的分子机制及临床意义研究

基本信息

批准号：
16601001
负责人：
YASUDA Osamu
金额：
$ 2.5万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16601001/
关键词：
Apoptosis Atherosclerosis Smooth muscle cell Bcl-2ファミリー PTP

项目摘要

Apoptosis (programmed cell death) of vascular smooth muscle cells (SMC) has been recognised recently in the vessel wall in disease states such as atherosclerosis and restenosis after angioplasty, and also in physiological arterial remodelling. The decrease of cells in atherosclerotic plaques by apoptosis contributes to the formation of unstable plaques, which are prone to rupture and trigger cardiovascular events. We have identified a novel gene, PL-3,in atherosclerotic smooth muscle cells of mice. Transient transfection of PL-3 expression vector induced apoptosis of cultured cells. To clarify the mechanism of PL-3-induced apoptosis, we performed Western blotting analysis, and found that cytochrome c is released from mitochondria into cytosolic space, followed by the activating caspase-9 and caspase-3. The cytochrome c release from mitochondria was inhibited by permeability transition pore (PTP) inhibitors, 1-carnitine or cyclosporin A, but not by bcl-2 or bcl-xL, anti-apoptotic Bcl-2 family proteins. Cyclophilin D (a component of PTP)-deficiency also prevented the apoptosis induced by PL-3. These data indicate that cytochrome c release from mitochondria induced by PL-3 expression is dependent on PTP rather than bcl-2 family-related channels. In order to clarify the physiological function of PL-3,we established experimental system of PL-3 inhibition using antisense plasmid or RNAi. Cell death induced by apoptosis-inducing reagents including lysophosphatidyl chorine, sodium nitroprusside or by hypoxia (1% oxygen), was prevented by the inhibition of PL-3 expression. This result indicates that PL-3 expression is implicated in cell death under physiological conditions. We have generated a PL-3-targeted ES cells to and introduced into the blastcysts to make a knockout mice of PL-3 gene. Furthermore, we have established an adenoviral expression system of PL-3 for the study gene therapy using rat models of carotid artery hypertrophy after balloon injury.

最近在动脉粥样硬化和血管成形术后再狭窄等疾病状态以及生理性动脉重塑中的血管壁中发现了血管平滑肌细胞（SMC）的凋亡（程序性细胞死亡）。动脉粥样硬化斑块中的细胞因凋亡而减少，导致不稳定斑块的形成，这些斑块容易破裂并引发心血管事件。我们在小鼠动脉粥样硬化平滑肌细胞中发现了一种新基因，PL-3。 PL-3表达载体瞬时转染诱导培养细胞凋亡。为了阐明PL-3诱导细胞凋亡的机制，我们进行了Western blotting分析，发现细胞色素c从线粒体释放到胞质空间，随后激活caspase-9和caspase-3。线粒体的细胞色素 c 释放受到通透性转换孔 (PTP) 抑制剂、1-肉碱或环孢菌素 A 的抑制，但不受抗凋亡 Bcl-2 家族蛋白 bcl-2 或 bcl-xL 的抑制。亲环蛋白 D（PTP 的一个组成部分）缺陷也阻止了 PL-3 诱导的细胞凋亡。这些数据表明 PL-3 表达诱导的线粒体细胞色素 c 释放依赖于 PTP 而不是 bcl-2 家族相关通道。为了阐明PL-3的生理功能，我们建立了利用反义质粒或RNAi抑制PL-3的实验体系。抑制 PL-3 表达可防止细胞凋亡诱导剂（包括溶血磷脂酰氯、硝普钠）或缺氧（1% 氧气）诱导的细胞死亡。该结果表明PL-3表达与生理条件下的细胞死亡有关。我们制备了靶向PL-3的ES细胞，并将其导入囊胚中，制成PL-3基因敲除小鼠。此外，我们还建立了PL-3的腺病毒表达系统，用于使用球囊损伤后颈动脉肥大的大鼠模型来研究基因治疗。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Fas Signaling Induces Akt Activation and Upregulation of Endothelial Nitric Oxide Synthase Expression

DOI：
10.1161/01.hyp.0000120124.27641.03
发表时间：
2004-04
期刊：
Hypertension: Journal of the American Heart Association
影响因子：
0
作者：
Y. Takemura;K. Fukuo;O. Yasuda;Takahito Inoue;N. Inomata;T. Yokoi;H. Kawamoto;T. Suhara;T. Ogihara
通讯作者：
Y. Takemura;K. Fukuo;O. Yasuda;Takahito Inoue;N. Inomata;T. Yokoi;H. Kawamoto;T. Suhara;T. Ogihara

Fas ligand mRNA 1 evels of circulating leukocytes reflect endothelial dysfunction in hyperlipidemic but not in non-hyperlipidemic patients

循环白细胞的 Fas 配体 mRNA 1 水平反映高脂血症患者的内皮功能障碍，但不反映非高脂血症患者的内皮功能障碍

DOI：
发表时间：
期刊：
Hypertension Research (in press)
影响因子：
0
作者：
YOSHIDA;TADAHIKO;(ed.);吉田忠彦(編著);Kawamoto H;Kotani N;Monta M;Toyohiko Yokoi;Kawamoto H;Kotani N;Tsubakimoto M;Suhara T;Takemura Y;Suhara T;Kawamoto H;Kotani N
通讯作者：
Kotani N

Timp-3 plays important roles in kidney following unilateral ureteral obstruction.

Timp-3 在单侧输尿管梗阻后的肾脏中发挥重要作用。