急性肺损伤微环境调控肺泡上皮细胞修复作用与机制研究

Repair and regeneration of alveolar tissue determines the prognosis of patients with ALI/ARDS. To enhance the repair ability of alveolar tissue is the most effective approach to solve the problem of ALI/ARDS treatment. Previously, we have found that: ① Alveolar epithelium has a strong self-repair ability after certain extent of ALI; ②Both lung tissue homogenate and bronchoalveolar lavage fluid from ALI lung tissues can promote the repair of alveolar epithelial cells, with homogenates showing more significant effects; ③In early-stage ALI homogenate AEC2 cells showed a certain degree of plasticity; ④After ALI, a variety of growth factors in lung tissue showed different patterns of changes. However, it remains unclear how the change of ALI microenvironment activates and regulates repair ability of alveolar epithelial cells. The current project aims to systematically study the Initiators and regulators of repair ability of AEC1 and AEC2 after ALI, to investigate the different regulation mechanisms of chemical microenvironments (lung homogenates) and physical microenvironments (bio-electric fields) on AEC1 and AEC2 behaviors, to systematically screen key factors regulating alveolar epithelial cell behaviors using a proteomics approach, and to study the molecular mechanisms for regulation of repair ability of alveolar epithelial cells by ALI microenvironments using signal transduction of protein chip technology. In summary, the current project aims to reveal both endogenous regulation mechanisms and key regulatory factors for alveolar self repair and regeneration after ALI, and to provide theoretical basis for finding measures to repair pulmonary tissue in patients with ALI/ARDS.

ALI后肺泡组织的修复与再生决定着ALI/ARDS患者的预后。我们前期研究发现：①一定程度ALI后肺泡上皮具有较强的自我修复能力；②ALI肺组织匀浆液和肺泡灌洗液均能促进肺泡上皮细胞修复，以匀浆液作用更明显；③ALI早期组织匀浆液可刺激AEC2细胞发生可塑性变化；④ALI后肺组织内多种生长因子呈不同的变化规律。然而，关于ALI微环境变化如何激活并调控肺泡上皮的修复，目前尚不清楚。本项目以ALI微环境为调控因素，系统研究AEC2和AEC1修复作用的启动与调控，探讨ALI微环境内化学与物理因素的不同调控作用，利用蛋白质组方法，系统筛选ALI微环境中调控肺泡上皮细胞的关键因子，利用信号转导技术，系统研究ALI微环境调控肺泡上皮修复作用的分子机制。旨在揭示ALI后肺泡自我修复与再生的内源性调控机制与关键调控因素，为解决ALI/ARDS的治疗难题提供理论基础与思路

肺脏不仅是重要的生命器官，也是各种危重疾病时最易受损的靶器官。因此，如何预防或/和有效治疗ALI/ARDS，对于提高严重创伤等危重疾病的救治水平至关重要。主要研究内容包括：①ALI后局部微环境变化对AEC2 细胞修复作用的不同调控与分子机制；②ALI后AEC1细胞可塑性变化及其与局部微环境变化的关系；③ALI后局部微环境中调控肺泡上皮修复的相关关键因子筛选与作用机制；④ALI后AEC1和AEC2 细胞的协同修复作用及其与局部微环境变化的关系。取得以下重要进展：①首次揭示ALI不同时间点肺匀浆液对AEC1、AEC2细胞的调控作用，发现早期匀浆液主要促进AEC2增殖和AEC1“去分化”，Wnt信号通路在其中发挥重要调控作用；②发现ALI晚期肺匀浆液可促进AEC2分化为AEC1，形成肺泡样结构，p38 MAPK是其重要的信号通路；③发现生物电场对肺泡上皮细胞有明显的调控作用，主要是促进AEC2、AEC1定向迁移，ERK1/2和PI3K/Akt是其重要的信号转导机制；④筛选出系列与ALI后肺组织自我修复作用密切关联的蛋白质分子，包括肝细胞生长因子、结缔组织生长因子、YAP1蛋白、髓样细胞触发受体-1等，并证实其不同的修复作用，为进一步探寻增强内源性肺修复措施提供了重要依据。⑤建立高效特异性CRISPR-Cas13 RNA编辑技术。共发表论文15篇，其中SCI论文12篇，影响因子>5的6篇。培养硕士、博士和博士后共9人。团队入选陆军重点扶持创新团队，团队成员获吴阶平医学创新奖、重庆市杰出英才奖，4人次入选省部级人才工程。近年来，团队分别与美国北达科大学和匹兹堡大学开展合作研究，在国际高影响力期刊联合发表SCI论文7篇，协助培养2名优秀青年。基于近年来的合作研究基础，今年与美国北达科大学联合申报获国家自然科学基金重点国际合作研究项目1项。应邀在国际学术会议报告6次，其中2次为特邀报告。