GLOMERULAR FUNCTION/BLOOD PRESSURE--PREGNANCY

肾小球功能/血压——怀孕

基本信息

批准号：
3086886
负责人：
Kirk P Conrad
金额：
$ 7.47万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-04-01 至 1990-03-31
项目状态：
已结题

项目摘要

The purpose of seeking an Individual Physician Scientist Award is to provide the applicant with research training in Phase 1, under the sponsorship of Dr. Michael J. Dunn (Case Western), that deals with in vitro techniques pertaining to the study of glomerular function. These techniques include: (a) isolation of rat glomeruli, (b) culture of rat glomerular epithelial and mesangial cells, (c) preparation of these cells for electronmicroscopy, and their subsequent identification, (d) analysis of basal and stimulated levels of cyclooxygenase products from freshly isolated glomeruli and glomerular cells in culture by radioimmunoassay and radiometric thin layer chromatography and (e) quantification of angiotensin II (AII) receptor density and affinity in the glomerulus. Additionally, the applicant will learn the extraction and chromatographic procedures required for purification of prostaglandins (PGs) from complex biological fluids. In Phase 2, under the sponsorship of Dr. Heinz Valtin (Dartmouth), these techniques will be applied to the study of glomeruli from pregnant rats, in order to discover a potential role for glomerular cyclooxygenase products, and angiotensin II receptors in the regulation of GFR, ERPF, and renal vascular resistance (RVR). Specifically, the applicant will examine in glomeruli isolated from pregnant rats (a) basal synthetic rates of PGs, (b) Michaelis-Menten kinetics of the cyclooxygenase system, (c) stimulated synthetic rates of PGs by addition of exogenous AII, arginine vasopressin, and norepinephrine, (d) potential production of 2,3-dinor-6keto-PGF1Alpha, (e) possible effects of estradiol and progesterone on PG production by cultured epithelial and mesangial cells, and (f) AII receptor density and affinity. These in vitro approaches will be carried out in conjunction with further studies in the chronically instrumented, conscious rat -- a method in which the applicant is currently expert, and has applied successfully to the investigation of pregnancy -- that utilize cyclooxygenase inhibition and infusions of angiotensin II and norepinephrine in an effort to discover a role for PGs in the modulation of GFR, ERPF, RVR and mean arterial pressure during pregnancy. In summary, this proposal addresses the potential role of PG/AII interactions in the control of glomerular function and blood pressure during pregnancy. A better understanding of the mechanisms that regulate the physiology of pregnancy would undoubtedly facilitate investigations of the pathogenesis of certain conditions that often complicate pregnancy -- e.g., preeclampsia and bilateral renal central necrosis.

寻求个人医师科学家奖的目的是在第一阶段为申请人提供研究培训， Michael J. Dunn 博士 (Case Western) 的赞助，从事体外研究与肾小球功能研究相关的技术。这些技术包括：(a) 分离大鼠肾小球，(b) 培养大鼠肾小球上皮细胞和系膜细胞，(c) 这些细胞的制备用于电子显微镜检查及其后续鉴定，(d) 分析新鲜环氧合酶产物的基础水平和刺激水平通过放射免疫分析分离肾小球和培养的肾小球细胞辐射薄层色谱法和 (e) 血管紧张素定量肾小球中 II (AII) 受体密度和亲和力。此外，申请人将学习提取和色谱程序从复杂的生物制品中纯化前列腺素 (PG) 所需的液体。在第二阶段，在 Heinz Valtin 博士（达特茅斯）的赞助下，这些技术将应用于妊娠期肾小球的研究大鼠，以发现肾小球环氧合酶的潜在作用产品和血管紧张素 II 受体调节 GFR、ERPF 和肾血管阻力（RVR）。具体来说，申请人将审查从怀孕大鼠分离的肾小球中 (a) PG 的基础合成率， (b) 环氧合酶系统的 Michaelis-Menten 动力学，(c) 受激通过添加外源性 AII、精氨酸加压素、和去甲肾上腺素，(d) 2,3-dinor-6keto-PGF1Alpha 的潜在产量， (e) 雌二醇和黄体酮对 PG 产生的可能影响培养的上皮细胞和系膜细胞，以及 (f) AII 受体密度和亲和力。这些体外方法将结合进行随着对长期使用仪器、有意识的老鼠的进一步研究—— 申请人目前是专家并已申请的方法成功地进行妊娠调查——利用环加氧酶抑制和血管紧张素 II 的输注去甲肾上腺素致力于发现 PG 在调节中的作用怀孕期间的 GFR、ERPF、RVR 和平均动脉压。总之，该提案阐述了 PG/AII 的潜在作用控制肾小球功能和血压的相互作用怀孕期间。更好地理解调节机制怀孕的生理学无疑将有助于研究某些常使妊娠复杂化的病症的发病机制—— 例如，先兆子痫和双侧肾中央坏死。