EFFECT OF CRANBERRY CONSTITUENTS ON UTI PATHOGENESIS

描述（由申请人提供）：尿路感染（uti）是最常见的细菌感染之一，其症状包括排尿困难、尿急和尿频、侧腹疼痛和发烧。临床诊断是基于症状和实验室发现的细菌尿。所有年龄段的妇女都特别容易受到急性和复发性感染。由于耐抗生素细菌的增加和长期抗生素预防的有害影响，需要对尿路感染进行替代治疗。治疗急性和复发性尿路感染的替代疗法包括摄入蔓越莓或蔓越莓汁。蔓越莓的作用包括酸化尿液、抑制激酶、抗炎、抗粘连和抗氧化作用。Vorsa实验室将蔓越莓果实分为主要的类黄酮类：原花青素、黄酮醇和花青素。通过生物测定定向分离，蔓越莓原花青素部分被证明含有可以抑制人类红细胞聚集的化合物，细菌表达PapG，与肾盂肾炎相关的细菌粘附素。此外，黄酮醇提取物在tpa诱导的小鼠耳部水肿模型中表现出较高的抗炎活性。Hultgren实验室利用小鼠膀胱炎模型来阐明参与尿路感染建立和持续的细菌和宿主因子的结构、功能和作用机制。在引入尿路致病性大肠杆菌（UPEC）时发生的致病级联反应的步骤已被描述。这个级联包括细菌结合和侵入膀胱上皮细胞，细胞内生长，流动，储层形成和再出现导致复发。宿主通过免疫应答应答，包括细胞因子的释放和pmn的浸润。在合作中，Vorsa和Hultgren实验室将研究特定蔓越莓成分的生物利用度及其对UPEC发病机制和宿主反应的每个步骤的影响。我们将利用化合物和蛋白质HPLC和FPLC纯化，亲和层析，小鼠模型，广泛的体外和组织培养模型，高分辨率EM， DNA微阵列，质谱，x射线晶体学和视频显微镜。这项工作将有助于了解蔓越莓中特定成分在UPEC发病机制中的生物活性，并可能为UPEC的治疗提供更好的方法。

DESCRIPTION (provided by applicant): Urinary tract infections (UTIs) are among the most common bacterial infections, with symptoms that can include dysuria, urgency and frequency of urination, flank pain and fever. Clinical diagnosis is based on symptoms and the laboratory finding of bacteriuria. Women of all ages are especially susceptible to acute and recurrent infections. Because of the rise in antibiotic-resistant bacteria and the deleterious effects of long-term antibiotic prophylaxis, alternative therapies for UTIs are needed. Alternative therapies proposed for treatment of acute and recurrent UTIs have included the ingestion of cranberries or cranberry juice. The actions ascribed to cranberries include acidification of the urine, inhibition of kinases, and anti-inflammatory, anti-adhesive and anti-oxidant effects. The Vorsa laboratory has fractionated cranberry fruit into the major flavonoid classes of proanthocyanidins, flavonols and anthocyanins. Through bioassay directed fractionation, the cranberry proanthocyanidin fraction was shown to contain compounds that can inhibit the aggregation of human erythrocytes by bacteria expressing PapG, the bacterial adhesin associated with pyelonephritis. In addition, the flavonol extract exhibited high anti-inflammatory activity in a TPA-induced mouse ear edema model. The Hultgren laboratory has utilized a mouse model of cystitis to elucidate the structure, function and mechanism of action of bacterial and host factors involved in the establishment and persistence of UTIs. The steps of the pathogenic cascade that occurs upon introduction of uropathogenic E. coli (UPEC) into the urinary tract has been delineated. This cascade includes bacterial binding and invasion into bladder epithelial cells, intracellular growth, fluxing, reservoir formation and reemergence leading to recurrence. The host responds by mounting an immune response that includes the release of cytokines and infiltration of PMNs. In collaboration, the Vorsa and Hultgren laboratories will investigate the bioavailability of specific cranberry constituents and their effects on each of the steps of UPEC pathogenesis and the host response. We will utilize compound and protein HPLC and FPLC purification, affinity chromatography, mouse models, extensive in vitro and tissue culture models, high resolution EM, DNA microarrays, mass spectroscopy, X-ray crystallography and videomicroscopy. This work will lead to a molecular under standing of the bioactivities of defined cranberry constituents on UPEC pathogenesis and may lead to better treatments for UTIs.