INHIBITION OF VASCULAR AND RENAL CELL PROLIFERATION

这是在抑制黄曲霉毒素诱导的细胞增殖方面的应用 心房肽(AP)和一氧化氮(NO)产生的药物。我们的目标是 根据我们的观察，AP和NO生成的血管扩张剂抑制 血清诱导培养的大鼠主动脉血管有丝分裂和增殖 肌肉和肾小球系膜细胞以及8-溴-cGMP模拟这些细胞 效果。这些结果支持了一个新的函数的存在性 血管活性激素：心钠素和血管紧张素转换酶的局部调节 血管紧张性：内皮源性一氧化氮(EDNO)。以下是 具体目标构成拟议实验的基础：目标1： 比较AP、NO-生成血管扩张剂和8-氨基丁酸的抗分裂作用 在原代和继代培养的主动脉平滑肌和 系膜细胞，以及除主动脉平滑肌和血管外的细胞类型 系膜细胞；我们建议使用培养的来自 冠状动脉、主动脉内皮细胞、肾上皮细胞NAD 3T3 成纤维细胞。目标2：确定受影响的细胞周期时相 APS、NO生成剂和8-bromo-cGMP的作用及其影响 这些抗分裂原对胸腺嘧啶核苷摄取的一个可能机制 行动。目的3：研究AP、NO生成的影响 血管扩张剂和8-溴-cGMP对促有丝分裂NAD细胞增殖的影响 通过确定的影响G1期早期或晚期事件的生长因素 将被测试的早期作用的有丝分裂原包括， 血小板衍生生长因子、碱性成纤维细胞生长因子和 而表皮生长因子是一种后期作用的有丝分裂原，将进行测试 是胰岛素样生长因子I(生长抑素C)。目标4：调查 AP、NO扩张剂和8-bromo-cGMP对大鼠血管紧张素转换酶活性的影响 由生长因子诱导的生化事件；其中包括调节 增加细胞内游离钙离子浓度、细胞pH、肌醇蓄积 磷酸盐和甘油三酯，以及原癌基因c-fos的表达。目标 5.评估上述措施的短期和长期效果 抗肿瘤药物对受体结合力、亲和力和受体数量的影响 增长因素。目的6：研究抗核素和抗核素 AP和NO生成的血管扩张剂的抗增殖作用是 仅由cGMP介导，还是cGMP不依赖的机制也 贡献力量。目的7：确定EDNO/内皮来源 松弛因子(EDRF)抑制血管平滑肌/系膜细胞 内皮细胞与血管共培养中的有丝分裂与增殖 平滑肌/系膜细胞。结果将提供有关 EDRF/NO、心钠素和心钠素的作用及机制 环鸟苷酸对血管平滑肌和系膜细胞的调节作用 体外有丝分裂和增殖。这些实验可能会促进 EDRF/EDNO和心房利钠激素发挥重要作用的概念 在维持血管平滑肌细胞有丝分裂静止中的作用 肾小球系膜细胞。

This is an application on the inhibition of cell proliferation induced by atriopeptins (APs) and nitric oxide (NO)-generating drugs. The aims are based on our observations that APs and NO-generating vasodilators inhibit serum-induced mitogenesis and proliferation of cultured rat aortic smooth muscle and renal mesangial cells and that 8-bromo-cGMP mimics these effects. These results support the existence of a novel function for a vasoactive hormone: atrial natriuretic hormone and a local regulator of vascular tone: endothelium derived nitric oxide (EDNO). The following specific aims form the basis of the proposed experiments: Aim 1: To compare the antimitogenic effects of APs, NO-generating vasodilators and 8- bromo-cGMP, in primary versus subcultured aortic smooth muscle and mesangial cells, and in cell types other than aortic smooth muscle and mesangial cells; we propose to use cultured smooth muscle cells from coronary artery, aortic endothelial cells, renal epithelial cells nad 3T3 fibroblasts. Aim 2: To identify the cell cycle phase that is affected by APs, NO-generating agents and 8-bromo-cGMP and to investigate the effects of these antimitogens on thymidine uptake a sa possible mechanism of action. Aim 3: To investigate the effects of APs, NO-generating vasodilators and 8-bromo-cGMP on mitogenesis nad cell proliferation induced by defined growth factors that affect early or late events in the G1 period of the cell cycle; early-acting mitogens that will be tested include, platelet-derived growth factor, basic fibroblast growth factor and epidermal growth factor whereas a later-acting mitogen that will be tested is insulin-like growth factor I (somatomedin C). Aim 4: To investigate the effects of APs, NO-generating vasodilators and 8-bromo-cGMP on various biochemical events induced by growth factors; these include the modulation of the increase of cytosolic free Ca2+, cell pH, accumulation of inositol phosphates and diglycerides, and expression of proto-oncogene, c-fos. Aim 5: To evaluate the short-term and long-term effects of the aforementioned antimitogens on receptor binding, affinity and number of receptor for growth factors. Aim 6: To investigate whether the antimitogenic and antiproliferative effects of APs nad NO-generating vasodilators are mediated by cGMP only, or whether cGMP-independent mechanisms also contribute. Aim 7: To establish whether EDNO/ endothelium-derived relaxing factor (EDRF) inhibits vascular smooth muscle/mesangial cell mitogenesis and proliferation, in cocultures of endothelial and vascular smooth muscle/mesangial cells. The results will provide information on the role and mechanism of action of EDRF/NO, atrial natriuretic peptides and cyclic GMP in the regulation of vascular smooth muscle and mesangial cell mitogenesis and proliferation in vitro. These experiments may promote the concept that EDRF/EDNO and atrial natriuretic hormone play an important role in maintaining the mitogenic quiescence of vascular smooth muscle and renal mesangial cells.