INHIBITION OF VASCULAR AND RENAL CELL PROLIFERATION

这是一个应用于抑制细胞增殖诱导的 心房肽（AP）和一氧化氮（NO）生成药物。 其宗旨是 基于我们的观察，AP和NO生成血管扩张剂抑制 血清诱导培养大鼠主动脉平滑肌细胞有丝分裂和增殖 肌肉和肾系膜细胞，8-溴-cGMP模拟这些 方面的影响. 这些结果支持存在一个新的功能， 血管活性激素：心房利钠激素和局部调节剂， 血管张力：内皮源性一氧化氮（EDNO）。 以下 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- 溴-cGMP，在原代与传代培养的主动脉平滑肌中， 系膜细胞，以及除主动脉平滑肌和 系膜细胞;我们建议使用培养的平滑肌细胞， 冠状动脉，主动脉内皮细胞，肾上皮细胞和3 T3 成纤维细胞 目的2：确定受以下因素影响的细胞周期阶段： AP、NO生成剂和8-溴环鸟苷（8-bromo-cGMP），并观察其作用 这些抗有丝分裂剂对胸苷摄取的一种可能的机制， 行动上 目的3：探讨AP、NO生成、 血管扩张剂和8-溴-cGMP对诱导的有丝分裂和细胞增殖的影响 通过影响G1期早期或晚期事件的特定生长因子 细胞周期;将被测试的早期作用有丝分裂原包括， 血小板衍生生长因子、碱性成纤维细胞生长因子和 表皮生长因子，而一种迟效有丝分裂原， 是胰岛素样生长因子I（生长调节素C）。 目标4：调查 AP、NO产生的血管扩张剂和8-溴-cGMP对各种 由生长因子诱导的生化事件;这些包括调节 细胞内游离Ca ~（2+）、细胞pH值、肌醇积累量的增加 磷酸盐和甘油二酯以及原癌基因c-fos的表达。 目的 5：评估上述措施的短期和长期影响 抗有丝分裂原对受体结合、亲和力和受体数目的影响 生长因子 目的6：研究抗有丝分裂和 AP和NO生成血管扩张剂的抗增殖作用是 是否仅由cGMP介导，或者是否cGMP非依赖性机制也 贡献. 目的7：确定EDNO/内皮源性 舒张因子抑制血管平滑肌/系膜细胞 有丝分裂和增殖，在内皮和血管的共培养物中 平滑肌/系膜细胞。 结果将提供有关 EDRF/NO、心房钠尿肽和 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 认为EDRF/EDNO和心钠素在心血管疾病中起重要作用， 维持血管平滑肌的促有丝分裂静止的作用， 肾系膜细胞

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.