P311在高氧肺损伤修复与重建中的作用及机制研究

Hyperoxia induced lung injury is a complex process that involving many cell activity. the final pathological changes of hyperoxia induced lung injury is alveolar development block and pulmonary interstitial fibrosis. How to improve the repair of lung injury are still poorly understood. Through extensive literature review and the results of our preliminary experiments that we find an important protein the P311. it plays an important role in the repair of lung injury. But its mechanism of action remains to be further explored. we propose the hypothesis: P311 may regulate lung cell proliferation and transdifferentiation regulated TGF-β1 / smad signal transduction pathways, reduce pulmonary fibrosis. To test this hypothesis, we will through the alveolar epithelial cells, pulmonary fibroblasts and high concentrations of oxygen induced bronchopulmonary dysplasia model of mouse , From the molecular, cellular, tissue and whole animal level to explore other aspects of the important role of P311 in hyperoxia-induced lung injury and repair and reconstruction. using real time PCR, Western blot, transfection of adenovirus vector, RNA interference and other means. In this study, taking P311 as the breakthrough point that from two cross disciplinaryperspective of the development and regeneration of science to reveal the mechanism of repair in lung injury, and to provide new ideas for the prevention and treatment of hyperoxia-induced lung injury.

高氧肺损伤是一个涉及许多细胞活动的复杂过程，其最终病理改变为肺泡发育阻滞和肺间质纤维化，迄今对如何改善肺损伤修复仍知之甚少。通过查阅大量文献及我们的预实验结果提示P311作为肺泡形成及对抗纤维增生的重要蛋白，在肺损伤修复中起重要作用。但其作用机制还有待进一步探讨。为此，我们提出假说：P311可能通过调节肺细胞增生及转分化，调控TGF-β1/smad信号转导途径，减轻肺纤维化。为验证这一假说，我们将通过肺泡上皮细胞、肺成纤维细胞和小鼠高浓度氧所致支气管肺发育不良模型，采用real time PCR、Western blot、腺病毒载体转染、RNA干扰等手段，从分子、细胞、组织以及动物整体水平等多方面探讨P311在高氧肺损伤修复与重建中的重要作用。本研究将从发育学和再生修复学两个交叉学科的角度，以P311为突破口，揭示肺损伤修复的发生机制，为高氧肺损伤的防治提供新思路。

高氧肺损伤是一个涉及许多细胞活动的复杂过程，其最终病理改变为肺泡发育阻滞和肺间质纤维化，迄今对如何改善肺损伤修复仍知之甚少。通过查阅大量文献我们发现P311作为肺泡形成及对抗纤维增生的重要蛋白，在肺损伤修复中起重要作用。本研究首先通过构建小鼠高浓度氧所致支气管肺发育不良动物模型，利用免疫组化、real time PCR、ELISA等实验方法，证明长时间吸入高浓度氧可成功构建支气管肺发育不良的动物模型，高氧肺损伤后肺组织P311 基因表达增加，P311 基因可能是影响肺损伤修复的调控因子；然后通过体外分离小鼠原代肺泡II型细胞及肺成纤维细胞，构建高氧肺细胞损伤模型，采用real time PCR、Western blot、RNA干扰等手段，从分子、细胞水平证实高氧可引起肺细胞生长不良，使AECⅡ迁移水平降低而肺成纤维细胞迁移增加，肺成纤维细胞向肌成纤维细胞转分化，肺细胞内ROS大量生成，最终引起肺细胞大量凋亡；肺细胞中P311的表达，随肺细胞凋亡水平升高而上调，推测该蛋白可能参与肺细胞凋亡的调控。.本项目从寻找促进肺组织细胞修复再生的调控因子P311基因为突破口，证实P311基因在肺损伤修复中起关键作用,其是肺组织细胞新的调控因子, 为P311基因促进肺损伤向无瘢痕愈合假设提供理论依据。