| Project/Area Number |
16601001
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
細胞死(アポトーシス)
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| Research Institution | Osaka University |
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Principal Investigator |
YASUDA Osamu Osaka University, Graduate School of Medicine, Assistant, 医学系研究科, 助手 (00372615)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUO keisuke Mukogawa Woman's University, School of Human Environmental Sciences, Professor, 生活環境学部, 教授 (40156758)
SHIMIZU Shigeomi Osaka University, Graduate School of Medicine, Associate Professor, 医学系研究科, 助教授 (70271020)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Apoptosis / Atherosclerosis / Smooth muscle cell / Bcl-2ファミリー / PTP |
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| Research Abstract |
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.
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