MECHANISMS OF CELL INJURY IN EXTRACORPOREAL SHOCK WAVE LITHOTRIPSY

体外冲击波碎石术（SWL）产生肾脏副作用 包括微血管和肾小管上皮的结构损伤 以及肾功能的改变 拟议研究的目标 目的是确定休克波（SW）细胞损伤的机制， 肾 声空化在SWL产生液体微射流能够 破坏细胞，并产生活性氧分子（ROM）， 有助于细胞损伤。 因此，SW产生的空化产生两个 可能导致细胞损伤的现象。 因此，我们提出两个假设， 解决细胞对这两种潜在细胞破坏特征的反应 SW处理的结果：1）冲击波引起的空化损伤细胞， 细胞器膜，和2）SWL细胞损伤发生的ROM介导的 机制 我们将描述即刻损伤与延迟损伤的特征， 如果膜损伤取决于空穴，由电子自旋确定 如果SW改变膜渗透性，则共振（ESR）将评估细胞 取决于膜完整性的功能参数将 确定线粒体损伤的程度和相关的细胞功能 细胞超微结构的改变。 同时，我们将确定 如果膜“稳定剂”降低细胞损伤的严重性。 我们 将通过ESR鉴定SWL产生的自由基种类并定位 它们的产生部位，表征抗氧化防御时的损伤 机制被改变，确定ROM清除程序是否减少或防止 损伤，并将评估线粒体损伤导致的改变， 细胞呼吸 此外，我们假设3）机械应力其他 比空化导致细胞损伤更严重 我们将测试这个想法， 当空化和应力被调节时评估损伤。 额外 研究将检查SW的物理参数的影响 细胞损伤的严重程度。 实验将在 肾小管（LLC-PK 1）和血管内皮细胞（VSMC）的培养细胞系模型 内皮细胞（HUVEC），以及分离的肾小管，纯化的肾 线粒体和质膜制备物。 我们的目标是为 建立改进的SWL所需的信息基础 这样可以减少或消除肾损伤。

Extracorporeal shock wave lithotripsy (SWL) produces renal side effects including structural damage to the microvasculature and tubular epithelium and alterations in kidney function. The objective of the proposed research is to determine the mechanisms of shock w ave (SW) cell injury in the kidney. Acoustic cavitation during SWL produces liquid microjets capable of disrupting cells, and generates reactive oxygen molecules (ROM) that may contribute to cell injury. Thus, SW-generated cavitation produces two phenomena that may cause cell injury. Therefore, we pose two hypotheses to address cellular response to these two potentially cell-disruptive features of SW treatment: 1) that shock wave-induced cavitation damages cell and organellar membranes, and 2) that SWL cell injury occurs by an ROM-mediated mechanism. We will characterize immediate versus delayed injury, determine if membrane damage is dependent upon cavitation, determine by electron spin resonance (ESR) if SWs alter membrane permeability, will assess cell functional parameters that are dependent on membrane integrity, will determine the extent of mitochondrial injury and correlate cell functional changes with alterations in cell ultrastructure. Also, we will determine if membrane"stabilizing agents" reduce the severity of cell injury. We will identify by ESR the free radical species produced by SWL and localize their site of production, characterize injury when anti-oxidant defense mechanisms are altered, determine if ROM scavengers reduce or prevent injury and will assess for mitochondrial injury resulting in alterations in cell respiration. Also, we hypothesize 3) that mechanical stress other than cavitation contributes to cell injury. We will test this idea by assessing injury when cavitation and stress are regulated. Additional studies will examine the influence of the physical parameters of SW delivery on the severity of cell injury. Experiments will be performed on cultured cell line models of the renal tubule (LLC-PK1) and the vascular endothelium (HUVEC), and on isolated kidney tubules, purified kidney mitochondria and plasma membrane preparations. Our goal is to contribute to the foundation of information necessary to establish improved SWL protocols so that renal injury is reduced or eliminated.