在研新药(HA1600)治疗急性肺损伤的机理

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are among the most devastating causes of fatality in critically ill patients. Smoke-induced severe ALI/ARDS is often fatal. During the last cycle of our grant, we discovered that: 1) HA1600, a new ALI drug candidate, can significantly increase lung SOCS1 expression. Smoke-induced apoptosis and inflammatory responses were reduced in the cells transfected with SOCS-1 expression plasmids.2) MiR-155 deficiency protects against smoke-induced lung injury. HA1600 can significantly decrease smoke-induced lung MiR-155 expression. 3) Smoke induces increased lung SOCS-1 expression in MiR-155-/- mice. 5) We hypothesize that HA1600 decreases smoke-induced apoptosis and inflammation via SOCS-1 by inhibiting MiR-155. We propose the following specific aims to investigate our hypothesis: 1) To elucidate that HA1600 suppresses DISC formation and inflammasome assembly in smoke-induced lung injury via SOCS-1; 2) To elucidate whether MiR-155 targets the SOCS-1 in smog-induced lung injury; 3) To determine that MiR-155 is involved in smoke-induced epithelial apoptosis through DISC formation; 4) To determine that MiR-155 is involved in smoke-induced inflammasome assembly; 5) To determine that HA1600 inhibits DISC formation and inflammasome assembly via SOCS1 by inhibiting MiR-155 in smoke-induced lung injury.

经上轮面上基金资助，我们发现：1）我们在研治疗ALI新药(HA1600)能明显增加肺SOCS1表达。细胞转染SOCS1表达质粒后，毒烟诱导的细胞凋亡和炎症反应减少；2）毒烟诱导的肺损伤在MiR155敲除鼠明显减少。HA1600能明显抑制毒烟诱导的肺MiR155表达；3）毒烟使MiR155敲除鼠肺SOCS1增加。我们推测：HA1600通过抑制MiR155促进SOCS1, 从而抑制毒烟诱导肺损伤。我们提出五个目标去检验我们的推测:1）确定HA1600通过SOCS1抑制毒烟诱导的肺DISC形成以及炎性小体组装；2）确定MiR155靶向作用SOCS1参与毒烟诱导的肺损伤；3）确定MiR155经DISC形成参与毒烟诱导的上皮细胞凋亡；4）确定MiR155参与毒烟诱导的炎性体形成；5）确定HA1600通过抑制MiR155促进SOCS1，进而抑制毒烟诱导的肺DISC形成以及炎性小体组装。

项目背景：吸入火灾产生的毒烟/日常生活中的雾霾能引起急性肺损伤（ALI）。ALI和急性呼吸道窘迫综合征（ARDS）是临床常见的呼吸系统急危重症。.主要研究内容：本项目确定microRNA-155靶向作用于SOCS-1参与毒烟诱导的肺损伤，确定长非编码RNA肺癌转移相关转录本1（lncRNA MALAT1）通过上调细胞因子信号抑制物-1（SOCS-1）参与在研新药（HA1600）对毒烟诱导的肺损伤。确定性别差异在低分子量透明质酸（LMMHA）诱导的急性肺损伤中的作用。.重要结果及关键数据：.（1）：毒烟处理后MiR-155表达升高：MiR-155的表达在WT小鼠吸入毒烟15 min后逐渐升高，于12小时达到最高值，随后出现下降趋势；.（2）：敲除MiR-155减轻小鼠毒烟诱导的ALI症状；.（3）：敲除MiR-155减轻小鼠肺部中性粒细胞聚集与炎性细胞因子（MPO、MIP-2及KC）的释放；.（4）：毒烟处理后，敲除MiR-155上调小鼠肺组织及原代中性粒细胞中SOCS-1的表达；.（5）：沉默SOCS-1增强敲除MiR-155小鼠毒烟处理后的原代中性粒细胞中的炎症反应；.（6）：气管内雾化HA1600减轻毒烟诱导的急性肺损伤及肺水肿；.（7）：HA1600抑制毒烟诱导的急性肺部炎症反应：毒烟诱导后，HA1600减轻小鼠肺泡灌洗液（BALF）中性粒细胞及总细胞的数量、总蛋白及炎症因子（IL-1β、IL-6与TNF-α）的水平；.（8）HA1600抑制毒烟诱导的LncRNA MALAT1表达的下降；.（9）肺组织过表达MALAT1减轻毒烟引起的肺损伤并可延长暴露于毒烟后小鼠的存活时间；.（10）过表达MALAT1上调毒烟处理后SOCS-1的表达水平；.（11）性别差异在LMWHA诱导的ALI中的影响：与雄性小鼠相比，雌性小鼠的肺损伤情况减轻、BALF总细胞及中性粒细胞数量及MPO表达水平下降、细胞因子（IL-6、CXCL-1、CXCL-2与TNF-α）表达水平下降；.（12）雌激素减轻LMWHA诱导的肺损伤；.科学意义：我们的研究表明。鼻喷HA1600能显著减少毒烟引起的死亡、肺细胞凋亡和炎症反应，这为治疗这些重大疾病提供了新的希望。