1.Taizhou municipal hospital;2.Department of Critical Care，Taizhou municipal hospital;3.Department of Blood Transfusion，Taizhou municipal hospital;4.Taizhou Municipal Hospital
目的 研究PI3K/Akt/mTOR信号通路在TRALI发病机制中的作用。方法 利用创伤后失血，再大量输血的方法构建输血相关急性肺损伤（TRALI）动物模型，通过HE染色检测大鼠肺组织形态学变化判断大鼠是否发生肺水肿。ELISA和qRT-PCR检测TRALI大鼠动物模型外周血/肺组织中TNF-α、IL-6和IL-1β蛋白和mRNA表达水平；Western blotting检测PI3K/Akt/mTOR信号通路相关蛋白及凋亡相关Bax、Bcl-2和Caspase3蛋白的表达水平。结果 TRALI大鼠肺泡组织结构严重受损，肺泡壁增厚，肺泡腔有粉色水肿液，炎细胞浸润，水肿明显；炎症性细胞因子TNF-α、IL-6和IL-1β在外周血和肺组织中表达水平明显增加（p<0.05）；PI3K/Akt/mTOR信号通路被活化，p-mTOR/mTOR表达明显增加，并抑制了凋亡蛋白Bax和Caspase 3的表达，增加了抗凋亡蛋白Bcl-2的表达（p<0.05）。结论 mTOR作为一个有潜力的药物靶点，由于其作用机制的复杂性，界定其发挥保护和损伤作用的确切时间靶点，选择最佳用药时间是临床防控TRALI发生发展的重要手段。
Objective: To study the effect of PI3K/Akt/mTOR signaling pathway on the pathogenesis of TRALI. Methods: Animal model was established by the method of trauma-blood loss-massive transfusion, and the pulmonary histopathological changes were detected by HE staining to determine whether the rats had pulmonary edema. The protein and mRNA expression levels of TNF-α, IL-6 and IL-1β in peripheral blood or lung tissues in TRALI rat models were detected by ELISA and qRT-PCR. The expression levels of PI3K/Akt/mTOR signaling pathway related proteins and apoptosis-related proteins Bax, Bcl-2 and Caspase3 were detected by western blotting. Results: Alveolar tissue structure was seriously damaged, the alveolar wall was thickened, there was pink edema fluid in the alveolar cavity, inflammatory cells infiltrated, edema was obvious in TRALI model rat. The expression levels of inflammatory cytokines TNF-α, IL-6 and IL-1β were significantly increased in peripheral blood and lung tissues (P<0.05); PI3K/Akt/mTOR signaling pathway was activated, the expression of p-mTOR/mTOR was significantly increased, and the expression of apoptotic protein Bax and Caspase 3 was inhibited, and the expression of anti-apoptotic protein Bcl-2 was increased (P<0.05). Conclusion: As a potential drug target, mTOR is an important means for clinical prevention and control of the occurrence and development of TRALI by defining the exact time target of its protective and damaging effects and selecting the optimal time of medication due to its complicated mechanism.