Yang Chen, Fu Cong, Wang Mengxue, Zheng Junbo, Gao Yang, Zhu Huiting, Li Haoxuan, Li Dongxu, Guo Lichen, Yu Bing, Dai Qingqing
Department of Anesthesia, the Obstetrics & Gynecology Hospital of Fudan University, Shanghai, People's Republic of China.
Department of Critical Care Medicine, The Obstetrics & Gynecology Hospital of Fudan University, Shanghai, People's Republic of China.
Immunotargets Ther. 2025 Apr 7;14:433-449. doi: 10.2147/ITT.S502925. eCollection 2025.
Recombinant antithrombin (rAT) has been shown to protect lungs from ARDS and modulate immune responses, but its anti-inflammatory mechanisms remain unclear. This study aimed to explore the immunomodulatory effects and mechanisms of rAT in LPS-induced ARDS mice.
ARDS mouse model was established by intraperitoneally administration of 20 mg/kg LPS. After 3 hours of LPS administration, rAT or PBS was injected intravenously. Lung injury, alveolar permeability, serum inflammatory cytokines, immune cell infiltration in lung tissue, and the proportion of Th17 were assessed 36 hours after rAT administration. The functional roles of the differential expressed genes (DEGs), obtained from LPS-induced ARDS mice treated with or without rAT, were analyzed by GO, KEGG and GSEA enrichment analysis. The activation of NF-κB and NLRP3 inflammasome was evaluated by Western blot and immunofluorescence staining.
We found that rAT alleviated lung injury, reduced pulmonary permeability, decreased serum inflammatory cytokines, and suppressed immune cell infiltration and NLRP3 inflammasome activation. Moreover, rAT decreased the proportion of Th17 cells in lung tissues and peripheral blood, downregulated IL17a expression, and inhibited NF-κB signaling pathway in lung tissues. Additionally, the administration of IL-17A diminished the efficacy of rAT in mitigating lung injury, suppressing the immune response, and inhibiting the activation of the NF-κB signaling pathway in LPS-induced ARDS mice.
The findings of this study suggest that rAT alleviates lung injury and suppresses inflammatory responses by inhibiting the IL17a/NF-κB signaling axis, suggesting that rAT may serve as a potential therapeutic agent for mitigating pulmonary inflammation and improving the prognosis of ARDS induced by sepsis. Furthermore, this study provides important research data and theoretical basis for the clinical translation and application of rAT.
重组抗凝血酶（rAT）已被证明可保护肺部免受急性呼吸窘迫综合征（ARDS）的影响并调节免疫反应，但其抗炎机制仍不清楚。本研究旨在探讨rAT对脂多糖（LPS）诱导的ARDS小鼠的免疫调节作用及其机制。
通过腹腔注射20 mg/kg LPS建立ARDS小鼠模型。在注射LPS 3小时后，静脉注射rAT或磷酸盐缓冲液（PBS）。在注射rAT 36小时后评估肺损伤、肺泡通透性、血清炎症细胞因子、肺组织中的免疫细胞浸润以及辅助性T细胞17（Th17）的比例。通过基因本体论（GO）、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA）对来自接受或未接受rAT治疗的LPS诱导的ARDS小鼠中获得的差异表达基因（DEG）的功能作用进行分析。通过蛋白质免疫印迹法和免疫荧光染色评估核因子κB（NF-κB）和NLRP3炎性小体的激活情况。
我们发现rAT减轻了肺损伤，降低了肺通透性，减少了血清炎症细胞因子，抑制了免疫细胞浸润和NLRP3炎性小体激活。此外，rAT降低了肺组织和外周血中Th17细胞的比例，下调了白细胞介素17A（IL-17A）的表达，并抑制了肺组织中的NF-κB信号通路。此外，在LPS诱导的ARDS小鼠中，给予IL-17A减弱了rAT在减轻肺损伤、抑制免疫反应和抑制NF-κB信号通路激活方面的疗效。
本研究结果表明，rAT通过抑制IL-17A/NF-κB信号轴减轻肺损伤并抑制炎症反应，这表明rAT可能作为一种潜在的治疗药物来减轻肺部炎症并改善脓毒症诱导的ARDS的预后。此外，本研究为rAT的临床转化和应用提供了重要的研究数据和理论依据。
