THE PATHOGENESIS OF CHRONIC SINUSITIS

描述（申请人摘要）：本提案的长期目标 是表征生理、生化和病理变化 发生在临床诊断为 慢性鼻窦炎这些假设是基于这样的观察， 氧化物在正常人的鼻子中以高浓度存在， 而鼻窦炎患者则没有。我们假设，在对病毒和/或 细菌感染后，窦内纤毛上皮细胞产生 细胞因子接下来是激活的炎症细胞的浸润， 主要是产生超氧化物的中性粒细胞。在独特的环境中， 窦，即高一氧化氮浓度，超氧自由基 优先与一氧化氮反应产生高细胞毒性的 过氧亚硝酸盐物种，而不是与其天然清除剂，超氧化物 歧化酶我们认为，随后的自由基级联损伤 纤毛上皮这些事件，加上鼻窦中的碎屑， 导致一氧化氮的合成减少和吸收增加， 慢性鼻窦炎为了验证这些假设，将研究鼻窦粘膜 对照组和接受择期手术的慢性鼻窦炎患者。 上颌窦导管插入术和各种 在不同流速下的NO浓度将允许确定 一氧化氮的产生和吸收率在原位，此外， 在这些过程中确定纤毛搏动频率（使用 专门设计的激光多普勒探测器）将证明这种关系 纤毛功能和NO浓度之间的关系。窦的体外研究 粘膜和窦灌洗液，使用张力计，NO代谢物分析， 酶学，将进一步表征这些过程。免疫组 切除的鼻窦组织和存档的组织石蜡块的染色将 允许识别NOS亚型及其在健康人中的分布， 以及酶的数量和性质 （使用高度特异性NOS的蛋白质印迹、Elisa测定和酶促技术） 拮抗剂）。此外，先前描述的技术将用于 确定硝基酪氨酸残基，亚硝基化蛋白质和标记物 凋亡对400个细胞相关RNA的cDNA阵列分析将进一步给出 慢性鼻窦炎的发病机制和结果的见解，从这些 分析将提示哪些特定的细胞信号传导剂需要进一步 通过ELISA定量。动物研究表明， NO诱导的细胞损伤和免疫复合物/自身免疫性疾病。理解 人类鼻窦中的这些过程有望提供新的精液信息 这可能导致新的治疗方法，更好的管理，甚至预防 慢性鼻窦炎

DESCRIPTION (Applicant's Abstract): The long-term objectives of this proposal are to characterize the physiological, biochemical and pathological changes that occur in the maxillary sinuses of individuals with clinically diagnosed chronic sinus disease. The hypotheses are based on the observation that nitric oxide is present in high concentrations in the nose of normal individuals but not in sinusitis patients. We hypothesize that, in response to a viral and/or bacterial infection, ciliated epithelial cells lining the sinus generate cytokines. There follows an infiltration of activated inflammatory cells, mainly neutrophils that generate superoxide. In the unique environment of the sinus, i.e. high nitric oxide concentration, the superoxide free radical preferentially reacts with nitric oxide to generate the highly cytotoxic peroxynitrite species rather than with its natural scavenger, superoxide dismutase. We suggest that the ensuing free radical cascade injures the ciliated epithelium. These events, together with the detritus in the sinuses, result in a decreased synthesis and increased absorption of nitric oxide in chronic sinus disease. To test these hypotheses, sinus mucosa will be studied from control and chronic sinusitis patients undergoing elective surgery. Catheritization of the maxillary sinus and the introduction of various concentrations of NO at varying flow rates will allow a determination of the rates of nitric oxide production and absorption in situ, Further, the determination of ciliary beat frequency during these procedures (using a specially designed laser Doppler probe) will demonstrate the relationship between ciliary function and NO concentrations. In vitro studies of sinus mucosa and sinus lavage fluids, using tonometry, NO metabolite analysis and enzymology, will further characterize these processes. Immunohistochemical staining of resected sinus tissues and archival tissue paraffin blocks will allow an identification of the NOS isoforms and their distribution in healthy and diseased tissues as well as the amounts and properties of the enzymes (Western blots, Elisa assay and enzymatic techniques using highly specific NOS antagonists). In addition, the techniques previously described will be used to determine nitro tyrosine resides, nitrosylated proteins and markers of apoptosis. CDNA array analysis for 400 cell related RNAs will give further insights into the pathogenesis of chronic sinusitis and results from these assays will suggest which specific cell signaling agents need further quantification by ELISA. Animal studies suggest a relationship between NO-induced cell injury and immune-complex/ autoimmune diseases. Understanding these processes in the human sinus promises to provide new seminal information that could result in new therapy, better management and even prevention of chronic sinus disease.