Relationshio pregenitor cells derived from bone marrow and graft intimal hyperplasia

骨髓来源的祖细胞与移植物内膜增生的关系

• 批准号: 15591336
• 负责人: KOBAYASHI Masayoshi
• 金额: $ 2.24万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591336/
• 关键词: 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

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 … More 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

基于LacZ小鼠颈动脉间置下腔静脉存在内膜增生的特点，我们制作了移植模型，但由于技术上的困难，不得不放弃。除此之外，我们继续用家兔制作移植物模型。（方法）家兔随机分为对照组和普伐他汀或匹伐他汀组，对照组喂饲市售兔饲料，普伐他汀组和匹伐他汀组分别喂饲市售兔饲料中普伐他汀或匹伐他汀钠10 mg/kg。在适当的麻醉下进行颈动脉间置-颈静脉移植。在植入自体静脉移植物后的每个时间点，在全身麻醉下收获静脉移植物。移植物的中间部分用于组织学和免疫组织化学研究。术后4周取静脉移植物，肉眼观察内膜增生情况。植入2周后，gr ...更多信息 用PCNA抗体染色。随机取8个视野，计数PCNA阳性细胞数和总细胞数，计算PCNA指数。为了评价细胞凋亡活性，我们采用TUNEL法检测细胞凋亡。将阳性细胞数除以计数的细胞总数定义为移植物的TUNEL指数。另外，购买人脐静脉内皮细胞（HUVEC）和血管平滑肌细胞（VSMC）。用1 μ M和30 μ M普伐他汀处理细胞24小时，用10%三氯乙酸收获细胞。将所得沉淀物用抗磷酸化MYPT-1、抗MBS抗体、抗RhoA和抗eNOS/NOS III型进行免疫印迹。（结果）实验结果表明，在正常胆固醇血症的家兔中，口服亲水性他汀类药物普伐他汀和匹伐他汀可抑制移植静脉移植后4周的增生，抑制移植静脉移植后2周的新生内膜细胞增殖和凋亡。此外，我们发现普伐他汀抑制Rho激酶活性，促进内皮细胞，而不抑制血管平滑肌细胞Rho激酶活性。（结论）亲水性他汀类药物通过内皮细胞选择性抑制Rho激酶，从而抑制移植静脉的晚期衰竭。此外，这些结果强烈支持临床使用亲水性他汀类药物预防旁路移植术后晚期移植物衰竭。少

项目成果

Ischemic intestinal involvement in a patient with Buerger's disease

布尔格病患者的缺血性肠道受累

• 发表时间: 2004
• 期刊: review series rheumatology 2
• 作者: Kanaoka Y;et al.;Kanaoka Y et al.;Kanaoka Y et al.;Masayoshi Kobayashi;Masayoshi Kobayashi
• 通讯作者: Masayoshi Kobayashi

Thrombosis and pulmonary embolism-prophylaxis-DVT treatment

血栓形成和肺栓塞-预防-DVT治疗

• 发表时间: 2004
• 期刊: Rinshosanhujinka 58
• 作者: Kanaoka Y;et al.;Kanaoka Y et al.;Kanaoka Y et al.;Masayoshi Kobayashi;Masayoshi Kobayashi;Masayoshi Kobayashi;小林昌義;Koji Yamamoto;Akihiko Kuzuya;Masayoshi Kobayashi;Masayoshi Kobayashi
• 通讯作者: Masayoshi Kobayashi

Gene therapy for vascular proliferative disease

血管增生性疾病的基因治疗

• 期刊: In Gene Therapy Frontier, Medical Review
• 作者: Kanaoka Y;et al.;Kanaoka Y et al.;Kanaoka Y et al.;Masayoshi Kobayashi;Masayoshi Kobayashi;Masayoshi Kobayashi;小林昌義;Koji Yamamoto;Akihiko Kuzuya;Masayoshi Kobayashi;Masayoshi Kobayashi;Koji Yamamoto;Akihiko Kuzuya;Hiroshi Banno
• 通讯作者: Hiroshi Banno

Long Term Outcome of Femoropopliteal Bypass for Claudication and Critical Ischemia

股腘绕道术治疗跛行和严重缺血的长期结果

• 发表时间: 2004
• 期刊: Asian Cardiovascular & Thoracic Annals 12
• 作者: Kanaoka Y;et al.;Kanaoka Y et al.;Kanaoka Y et al.;Masayoshi Kobayashi
• 通讯作者: Masayoshi Kobayashi

Healing of implanted expanded polytetrafluoroethyrene vascular access grafts different in ternodel distances : a histologic study in dogs

不同节距的植入扩张聚四氟乙烯血管通路移植物的愈合：狗的组织学研究

• 发表时间: 2004
• 期刊: European Journal of Vascular and Endovascular Surgery 28
• 作者: Kanaoka Y;et al.;Kanaoka Y et al.;Kanaoka Y et al.;Masayoshi Kobayashi;Masayoshi Kobayashi;Masayoshi Kobayashi;小林昌義;Koji Yamamoto;Akihiko Kuzuya
• 通讯作者: Akihiko Kuzuya