EFFECT OF CRANBERRY CONSTITUENTS ON UTI PATHOGENESIS

描述(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、腰部疼痛和发烧。临床诊断是根据症状和实验室发现的菌尿。所有年龄段的妇女都特别容易受到急性和复发性感染。由于抗药性细菌的增加和长期抗生素预防的有害影响，尿路感染的替代疗法是必要的。治疗急性和复发性尿路感染的替代疗法包括摄取小红莓或小红莓汁。小红莓的作用包括酸化尿液，抑制酶，以及抗炎、抗粘连和抗氧化作用。沃萨实验室已经将蔓越莓果实分离成主要的类黄酮类，如原花青素、黄酮醇和花青素。通过生物测定定向分级，红莓原花青素部分含有能够抑制表达PapG的细菌对人红细胞聚集的化合物，PapG是一种与肾盂肾炎相关的细菌粘附素。此外，黄酮醇提取物在TPA诱导的小鼠耳肿胀模型中显示出较高的抗炎活性。Hultgren实验室利用膀胱炎的小鼠模型来阐明与尿路感染的建立和持续有关的细菌和宿主因素的结构、功能和作用机制。尿路致病性大肠埃希氏菌(UPEC)进入尿路时发生的致病性级联反应的步骤已被描述。这一级联反应包括细菌与膀胱上皮细胞的结合和侵入、细胞内生长、粘连、储存库的形成和导致复发的复发。宿主的反应是启动一种免疫反应，包括细胞因子的释放和中性粒细胞的渗透。在合作中，Vorsa和Hultgren实验室将调查特定蔓越莓成分的生物利用度及其对UPEC发病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高效液相色谱和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、质谱学、X射线结晶学和视频显微镜。这项工作将有助于从分子水平了解小红莓成分在UPEC发病机制中的生物活性，并可能导致更好地治疗尿路感染。

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.