Nitrite dependent protection against Cl2 gas toxicity_role of chlorinated lipids

描述(申请人提供)：氯(Cl2)气体介导的损伤是复杂的，涉及暴露期间发生的直接毒性和暴露后数小时对呼吸道、肺和全身血管系统的强烈毒性。初步数据和我们最近发表的研究表明，Cl2暴露后的损伤包括NO生物利用度的显著下降，其指标是肺和肺外血管中eNOS依赖的血管扩张功能减弱，并证明Cl2暴露后通过单一IM注射亚硝酸盐(一种无复制策略)，可以预防急性肺损伤、呼吸道高反应性和重要的死亡率。在这项提案中，我们将：i)建立一个治疗框架，用于开发亚硝酸盐作为暴露后治疗的药物，可在大规模伤亡情况下使用。Cl2后气体损伤是如何发生的尚不清楚。我们发现新的数据表明，Cl2气体增加了肺和循环中的氯化脂肪(CL-LIP)，这是源于血浆原的�-氯脂肪酸。CL-LIP可作为Cl2暴露的新生物标志物，但我们也提供证据表明，这些物种本身可导致急性肺损伤和呼吸道上皮细胞功能障碍。重要的是，氯-LIP依赖的毒性也可以通过暴露后亚硝酸盐治疗来预防。综上所述，我们假设氯-LIP是Cl2后气体毒性的新的生物标志物和媒介，暴露后亚硝酸盐治疗将减轻Cl2气体和氯-LIP依赖的毒性，并将通过以下特定目的进行验证：1.确定氯-LIP在介导Cl2气体诱导的毒性中的作用；2.确定减轻致死性和亚致死性亚硝酸盐的最佳治疗条件 Cl2气体毒性，3.确定GLP下IM亚硝酸盐给药的安全性/毒理学特征 条件。建议将体内Cl2气体暴露与体外研究相结合。我们认为这项研究既及时又重要，预计将对Cl2气体毒性的分子机制产生新的见解，并确定和开发治疗(对策)策略。

DESCRIPTION (provided by applicant): Chlorine (Cl2) gas mediated injury is complex involving a direct toxicity that occurs during the exposure and a robust post exposure toxicity that occurs over hrs-days to the airways, pulmonary and systemic vasculature. Preliminary data and our recently published studies show that injury post Cl2 exposure includes significant decreases in NO-bioavailability as indexed by diminution of eNOS-dependent vasodilation in the pulmonary and extrapulmonary vasculature and demonstration that post Cl2 exposure administration of nitrite (an NO- repleting strategy) by a single IM injection, protects against acute lung injury, airway hyper-reactivity and importantly, mortality. In this proposal we will i) establish a therapeutic framework for the development for the use of nitrite as a post exposure therapeutic that can be administered in mass casualty scenarios. How post- Cl2 gas injury occurs is unclear. We show novel data that Cl2 gas increases lung and circulating chlorinated lipids (Cl-lip), which are �-chloro fatty acids derived from plasmalogens. Cl-lip may serve as novel biomarkers for Cl2 exposure, but we also provide evidence that these species can themselves cause acute lung injury and airway epithelial cell dysfunction. Importantly, Cl-lip dependent toxicity can also be prevented by post-exposure nitrite therapy. Taken together, we hypothesize that Cl-lip are novel biomarkers and mediators of post Cl2 gas toxicity and that post-exposure nitrite therapy will attenuate Cl2 gas and Cl-lip dependent toxicity and will test this via the following Specific Aims: 1. Determine the role of Cl-lip in mediating Cl2 gas induced toxicity, 2. Determine optimal nitrite therapeutic conditions for attenuating lethal and sub-lethal Cl2 gas toxicity, 3. Determine safety / toxicology profiles for IM nitrite administration under GLP conditions. A combination of in vivo Cl2 gas exposure with ex vivo studies is proposed. We feel this research is both timely and important and anticipate will yield novel insights into molecular mechanisms of Cl2 gas toxicity and identify and develop therapeutic (countermeasure) strategies.