PATHOGENIC MECHANISMS IN HUMAN URINARY TRACT CELLS

慢性泌尿道（UT）疾病的病因，间质性 膀胱炎，是模糊的。 拟议研究的目的是 通过培养正常人，建立合适的UT上皮细胞模型， 血清中的人膀胱和/或输尿管尿路上皮（UE）细胞， 自由培养基。 我们还建议采用这种新的细胞模型， 研究细菌-UE细胞相互作用对几种 影响上皮细胞能力的不同参数 配体-受体结合信号。 我们计划研究 蛋白激酶-C（PK-C）的激活/抑制，以及 鸟氨酸脱羧酶（ODC）的诱导/抑制 并且在UE小区的扰动之后。 另外的实验 设计方案来评估外源性细胞的作用， 信号调制器（例如，细菌粘附，细菌和尿 代谢物，细菌脂多糖）对 蛋白聚糖 实验方案旨在 探讨表皮生长因子和类固醇对 激素（醛固酮、17-β-雌二醇、睾酮和 孕酮）对信号转导酶ODC和PK-C的影响，以及 对UE细胞与病原菌结合的感受性的影响 （大肠杆菌、奇异变形杆菌和金黄色葡萄球菌）。 最后，我们建议研究细菌的影响， 粘附、生长因子、激素和环磷酰胺 尿代谢产物丙烯醛对尿上皮细胞离子转运的影响 文化 特别地，实验方案被设计成 研究阳离子（Na+，K+）和阴离子（Cl-， SO 42-）通过顶侧和基底侧表面的摄取/流出 的UE小区。 我们相信这些不同的实验方案 应该为致病机制提供新的见解， 导致正常人类UE细胞的功能障碍， 膀胱和UT。

The etiology of the chronic urinary tract (UT) disease, interstitial cystitis, is obscure. The objective of the proposed research is to develop a suitable UT epithelial cell model by culturing normal human bladder and/or ureter uroepithelial (UE) cells in a serum- free medium. We also propose to employ this novel cell model to investigate the effect of bacterial-UE cell interactions on several different parameters affecting the capacity of the epithelial cells to transduce ligand-receptor binding signals. We plan to study the activation/inhibition of protein kinase-C (PK-C), and the induction/suppression of ornithine decarboxylase (ODC) before and after perturbation of the UE cells. Additional experimental protocols are designed to evaluate the effect of extrinsic cell signal modulators (e.g., bacterial adherence, bacterial and urinary metabolites, bacterial lipopolysaccharides) on the biosynthesis of proteoglycans. Experimental protocols are designed to investigate the effect of epidermal growth factor and steroid hormones (aldosterone, 17-beta-estradiol, testesterone, and progesterone) on signal-transducing enzymes, ODC and PK-C, and on the receptivity of UE cell binding to pathogenic bacteria (Escherichia coli, Proteus mirabilis, and Staphylococcus aureus). Lastly, we propose to investigate the effect of bacterial adherence, growth factors, hormones and the cyclophosphamide urinary metabolite, acrolein, on ion transport of UE cells in culture. In particular, experimental protocols are designed to study the regulation of both cation (Na+, K+), and anion (Cl-, and SO42-) uptake/efflux through the apical and basolateral surfaces of UE cells. We believe that these varied experimental protocols should provide new insights into the pathogenic mechanisms contributing to dysfunction of normal human UE cells of the bladder and UT.