| Project/Area Number |
15591336
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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 |
General surgery
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| Research Institution | Nagoya University |
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Principal Investigator |
KOBAYASHI Masayoshi Nagoya University, University Hospital, research associate, 医学部附属病院, 助手 (60329381)
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| Co-Investigator(Kenkyū-buntansha) |
KOMORI Kimihiro Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (40225587)
MUROHARA Toyoaki Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (90299503)
EMI Nobuhiko Nagoya University, Graduate School of Medicine, associate Professor, 大学院・医学系研究科, 助教授 (30185144)
錦見 尚道 名古屋大学, 医学部附属病院, 講師 (40242862)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | intimal hyperplasia / pregenitor cells / perlipheral vascular disease / statin / prevention of intimal hyperplasia / eNOS / 内膜肥圧抑制 / 頸動脈自家静脈バイパス / プラバスタチン |
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| Research Abstract |
Based on that intimal hyperplasia is recognized in the interposed inferior vena in the carotid arteries of LacZ mouse, graft model was made, However, it was very difficult to performe technically and we could not help giving it up. Otherwise we kept on making graft model with the use of rabbit. (Methods) Rabibits were randomly divided into two groups ; the control group that was fed commercial rabbit chow, and the pravastatin or pivatastatin group that received the same chow with 10mg/kg pravastatin or pivatastatin sodium. Carotid interposition-reversed juglar vein grafts were performedunder appropriate anesthesia. At each time point after implantation of the autologous vein grafts, the vein grafts were harvested under general anesthesia. The middle portion of the graft was used for histological and immunohistochemical studies. Four weeks after operation, vein grafts in both groups were harvested and intimal hyperplasia were elucidated macroscopically. After 2 weeks of implantation, gr
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aft were harvested and stained with PCNA antibodies. To calculate this PCNA index, PCNA positive cells and total cells were ciunted at random 8 fields. In order to evaluate apoptosis activity, we used the TUNEL method to detect in situ apoptosis. The number of positive cells divided by the total number of cells counted was defined as the TUNEL index of the graft. Additionally human umbilical vein endothelial cells(HUVECs) and vascular smooth muscle cells(VSMCs) was purchased. The cells were treated with 1-μ Mand 30-μM pravastatin for 24 hours and harvested with 10% trichloroacetic acid. The resulting precipitates were subjected to immunoblotting with anti-phospho-MYPT-1,anti-MBS antibody, anti-RhoA, and and anti-eNOS/NOS type III. (Results) We demonstrated that oral administration of the hydrophikic statin, pravastatin and pitavastatin, to normocholesterolemic rabbits inhibited hyperplasia of the vein grafts 4 weeks after implantation, and suppressed cell proliferation and apoptosis in the neointima 2 weeks after implantation. In addition, we found that pravastatin inhibited Rho-kinase activity and accerated endothelial cells, while it did not inhibit Rho-kinase activity in vascular smooth muscle cells. (Conclusion) Based on our findings hydrophilic statin can suppress late vein graft failure through endothelial cell-selective inhibition of Rho-kinase. Furthermore, these results strongly support the clinical use of hydrophilic statins to prevent late graft failure after bypass grafting. Less
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