Project 3: Anti-Inflammatory Mechanisms of Inhaled Hypertonic Saline

外伤、出血和血液复苏通过诱导趋化因子、细胞因子和粘附蛋白而引起急性肺部炎症。上调机制依赖于将基因调节转录因子（tf）（如NF-kB）转运到细胞核中的系统信号。这些基因直接表达了肺泡环境中白细胞的粘附、迁移和表型发展。高渗透压（HOsm）似乎可以阻止炎症基因表达，但其机制和范围尚不清楚。创伤复苏期间的治疗性高渗生理盐水输注，或吸入治疗囊性纤维化，耐受性良好。我们假设高渗透压通过促进新的复合物改变炎症刺激诱导的转录因子的核易位。在本提案期内（2005-2010），我们将首先研究控制HOsm修饰的原型肺泡细胞因子和粘附蛋白的tf的流量，并研究这些tf的复合物，包括与进口蛋白形成的复合物。

Trauma, hemorrhage and blood resuscitation produce acute lung inflammation by inducing chemokine, cytokine and, and adhesive proteins. Up-regulating mechanisms depend on systemic signals that translocate gene-regulating, transcription factors (TFs), such as NF-kB into the nucleus. The genes expressed direct the adhesion, transmigration of leukocytes and phenotypic developments in the alveolar milieu. Hyperosmolarity (HOsm), appears to prevent inflammatory gene expression but the mechanisms and scope are unclear. Therapeutic hypertonic saline infusions during trauma-resuscitation, or inhaled as for cystic fibrosis, are well tolerated. We hypothesize that hyperosmolarity alters the nuclear translocation of selected transcription factors induced by inflammatory stimuli by promoting novel complexes. In this proposal period (2005-2010) we will first examine the traffic of TFs controlling prototypical alveolar cytokines and adhesive proteins that are modified by HOsm, and investigate complexes of these TFs, including those formed with importins. 1. Investigate HOsm altered translocation of the NF-KB and IKK family members, their HOsm altered cytoplasmic complexes and examine alternate complexes promoted by HOsm with IP-MS proteomics. 2. Since HOsm does not prohibit p65 Rel A translocation induced by IL-1 yet suppresses certain cytokines, we will examine translocation of other essential promoters of the IRF, C/EBP. and R/FLAT families and identify their HOsm altered partners. Next, to expand the inquiry beyond the known cytokines we will execute: 3. Gene array analyses, to guide bioinformatics queries for potential TFs. The goal is to gradually build up a library of TFs that do not translocate successfully during HOSm and identify their sequestering partners, (starting with NF-kB and IRF family members). Lastly, we will translate these bench experiments to asses the tolerability or benefit of nebulized HOsm in shocked animals and trauma patients. 4. Evaluate lung inflammation and TF translocation in traumatized Animal or Patients treated with inhaled HOsm. The information from Aims 1-3 will be used to interpret the molecular phenotypes and TF redistributions after treatments.