Weng Junting, Weng Shuoyun, Xu Jitao, Liu Danjuan, Guo Rongjie, Shi Bingbing, Chen Min
Department of Clinical Medicine, Fujian Medical University, Fuzhou, 350000, China.
Department of Critical Care Medicine, the Affiliated Hospital of Putian University, No. 999 Dongzhen East Road, Licheng District, Putian, 351100, China.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Jan 11. doi: 10.1007/s00210-025-03786-9.
Acute lung injury (ALI) is a severe inflammatory condition of the respiratory system, associated with high morbidity and mortality. This study investigates the therapeutic potential of tocilizumab (TZ), an IL-6 receptor inhibitor, in mitigating lipopolysaccharide (LPS)-induced ALI by modulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway. An ALI model was established using LPS induction. Lung damage was assessed by hematoxylin-eosin (H&E) staining, alongside measurements of respiratory function, including PaO/Fio ratios, lung edema, airway resistance, and lung compliance. Western blotting analyzed the expression of phosphorylated PI3K and AKT (P-PI3K, P-AKT), while ELISA quantified levels of TNF-α, IL-1β, IL-6, and oxidative stress markers. Apoptosis was evaluated using the TUNEL assay, and key apoptotic proteins (Bcl-2, Bax, and caspase-3) were measured by Western blotting. The Cell Counting Kit-8 (CCK-8) assay was employed to determine cell viability. The LPS-induced model exhibited decreased P-PI3K and P-AKT levels, while TZ treatment significantly elevated these markers. TZ also reduced lung tissue damage, improved respiratory function, and decreased inflammation, oxidative stress, and apoptosis. However, co-administration with LY294002 (a PI3K inhibitor) blocked these benefits, reversing the protective effects of TZ. TZ alleviates lung injury and improves outcomes in LPS-induced ALI by enhancing the PI3K/AKT pathway.
急性肺损伤（ALI）是一种严重的呼吸系统炎症性疾病，具有高发病率和死亡率。本研究调查了白细胞介素-6受体抑制剂托珠单抗（TZ）通过调节磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）途径减轻脂多糖（LPS）诱导的ALI的治疗潜力。使用LPS诱导建立ALI模型。通过苏木精-伊红（H&E）染色评估肺损伤，同时测量呼吸功能，包括动脉血氧分压/吸入氧分数（PaO₂/FiO₂）比值、肺水肿、气道阻力和肺顺应性。蛋白质免疫印迹法分析磷酸化PI3K和AKT（P-PI3K、P-AKT）的表达，而酶联免疫吸附测定（ELISA）定量肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和氧化应激标志物的水平。使用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）法评估细胞凋亡，并通过蛋白质免疫印迹法测量关键凋亡蛋白（Bcl-2、Bax和半胱天冬酶-3）。采用细胞计数试剂盒-8（CCK-8）法测定细胞活力。LPS诱导的模型显示P-PI3K和P-AKT水平降低，而TZ治疗显著提高了这些标志物的水平。TZ还减少了肺组织损伤，改善了呼吸功能，并减轻了炎症、氧化应激和细胞凋亡。然而，与LY294002（一种PI3K抑制剂）联合使用可阻断这些益处，逆转TZ的保护作用。TZ通过增强PI3K/AKT途径减轻LPS诱导的ALI中的肺损伤并改善预后。
