DETERMINATION OF DNA SEQUENCE OF NEUROPEPTIDE Y Y3-RECEPTOR

神经肽 Y Y3-受体 DNA 序列的测定

• 批准号: 12670097
• 负责人: ISHIKAWA Naohisa
• 金额: $ 0.58万
• 依托单位: Aichi Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670097/
• 关键词: neuropeptide Y; neuropeptide Y Y3-receptor; cloning; lung vascular permeability; peptide YY; cDNA library; vascular endothelial cell; pulmonary circulation; neuropeptide Y Y_3受容体

In order to investigate a mechanism involved in the development of neurogenic pulmonary edema, we utilized three experiments: (1) cloning of neuropeptide Y Y3-receptor and determination of its DNA sequenc* (2) involvement of vascular endothelial cells; (3) action of central nervous system.We have reported that neuropeptide Y is a neurotransmitter, released from the sympathetic nerve terminal, which might increa* the pulmonary vascular permeability in the development of fibrin-induced pulmonary edema, a model of neurogenic pulmonary edema* So far, DNA sequence of NPY Y3-receptor has not been elucidated. Therefore it disturbs a generation of new drug. We extracted mR* and created cDNA. The cDNA was ligated to ZAP Express Vector, creating cDNA library. E.coli DNA was infected by phage which h* the cDNA. After incubation, E. coli successfully made plaques on agar plate. Blotted nitrocellulose membrane was treated with ant* NPY antibody and anti PYY antibody. Visualized with colorization, c … More loning is now under study.Secondly, we utilized rat aortic endothelial cells, making the cells alive in agar plates. Endothelial cells are now investigated f* detecting the vascular permeability with permeating molecules such as fluorescent molecule-binding albumin. At the same time, we * also studying the development of vascular endothelial cell growth factor (VEGF) which makes the vascular bed permeable to large * molecule, albumin. Venous anesthetics, ketamine, propofol, and thiopental, neurotransmitters, neuropeptide Y, norepinephrine and * acetylcholine, and other bioactive substances. Ketamine and thiopental have an activity to increase the release ofVEGF from the * vascular endothelial cells.We already reported that NO release in the medulla oblongata has enhanced the development of neurogenic pulmonary edema. * Since unilateral left-vagotomy increase the NO synthase 2 weeks later, we have examined the fibrin-induced pulmonary edema, obtain * the results demonstrating the inhibitory action of NO on the edema development. However, the increase in vascular permeability was * maximum at 2 weeks time point, but rather increased more at 4 weeks. Now we are studying the effects of 1-glutamate, an excitable * neurotransmitter, on the fibrin-induced pulmonary edema. Less

为了探讨神经源性肺水肿的发生机制，我们通过三个实验：(1)神经肽yy3受体的克隆及其DNA序列的测定；(3)中枢神经系统的作用。我们报道神经肽Y是交感神经末梢释放的一种神经递质，在纤维蛋白诱导的肺水肿（一种神经源性肺水肿模型）的发展过程中可能增加肺血管通透性*，但迄今为止，NPY y3受体的DNA序列尚未明确。因此，它扰乱了一代新药。提取mR*，构建cDNA。将cDNA连接到ZAP Express Vector上，建立cDNA文库。大肠杆菌DNA被噬菌体感染，噬菌体与cDNA结合。孵育后，大肠杆菌成功地在琼脂板上形成斑块。用蚂蚁* NPY抗体和抗PYY抗体处理印迹硝化纤维素膜。更多的孤独正在研究中。其次，我们利用大鼠主动脉内皮细胞，使细胞在琼脂板上活。内皮细胞现在被研究用渗透分子如荧光分子结合白蛋白检测血管通透性。同时，我们也在研究血管内皮细胞生长因子（VEGF）的发展，它使血管床能够渗透大分子白蛋白。静脉麻醉剂、氯胺酮、异丙酚、硫喷妥钠、神经递质、神经肽Y、去甲肾上腺素、乙酰胆碱等生物活性物质。氯胺酮和硫喷妥钠具有增加血管内皮细胞中vegf释放的活性。我们已经报道过延髓内NO的释放促进了神经源性肺水肿的发展。*由于单侧左迷走神经切断术2周后增加NO合成酶，我们检查了纤维蛋白诱导的肺水肿，得到*结果表明NO对水肿发展有抑制作用。而血管通透性的增加在2周时最大，在4周时增加更多。目前我们正在研究可兴奋性神经递质1-谷氨酸对纤维蛋白诱导肺水肿的作用。少

项目成果

Ishikawa,N. et al.: "Role of central nervous system nitric oxide in the development of nuerogenic pulmonary edema in rats"Clitical Care Medicine. (in press).

石川，N.

Ossama Hamdy, Kimitoshi Nishiwaki, Michi Yajima, Hidetsugu, 0. Murakami, Hiroshi Maekawa, Robert T. Moy, Yasuhiro Shimada, Naohisa Ishikawa: "Presence and quantification of neuropeptide Y in pulmonary edema fluids in rats"Experimental Lung Research. 26. 1

Ossama Hamdy、Kimitoshi Nishiwaki、Michi Yajima、Hidetsugu、0. Murakami、Hiroshi Maekawa、Robert T. Moy、Yasuhiro Shimada、Naohisa Ishikawa：“大鼠肺水肿液中神经肽 Y 的存在和定量”实验性肺研究。

Ossama Hamdy: "Presence and quantification of neuropeptide Y in pulmonary edema fluids in rats"Experimental Lung Research. 26. 137-147 (2000)

Ossama Hamdy：“大鼠肺水肿液中神经肽 Y 的存在和定量”实验性肺研究。

Ossama,Hamdy et al.: "Presence and quantification of neuropeptide Y in pulmonary edema fluid in rats"Experimental Lung Research. 26. 137-147 (2000)

Ossama、Hamdy 等人：“大鼠肺水肿液中神经肽 Y 的存在和定量”实验性肺研究。

Ossama Hamdy: "Role of central nervous system nitric oxide in the development of neurogenic pulmonary edema in rats"Crit Care Med. 29. 1222-1228 (2001)

Ossama Hamdy：“中枢神经系统一氧化氮在大鼠神经源性肺水肿发展中的作用”Crit Care Med。