Acute ethanol intoxication increases the risk of sepsis and aggravates the symptoms of sepsis and lung injury. Therefore, this study aimed to explore whether sphingosine kinase 1 (SphK1)/sphingosine-1-phosphate (S1P)/S1P receptor 1 (S1PR1) signaling pathway functions in lung injury caused by acute ethanol intoxication-enhanced sepsis, as well as its underlying mechanism. The acute ethanol intoxication model was simulated by intraperitoneally administering mice with 32% ethanol solution, and cecal ligation and puncture (CLP) was used to construct the sepsis model. The lung tissue damage was observed by hematoxylin-eosin (H&E) staining, and the wet-to-dry (W/D) ratio was used to evaluate the degree of pulmonary edema. Inflammatory cell counting and protein concentration in bronchoalveolar lavage fluid (BALF) were, respectively, detected by hemocytometer and bicinchoninic acid (BCA) method. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and IL-18 in BALF were detected by their commercial enzyme-linked immunosorbent assay (ELISA) kits. The myeloperoxidase (MPO) activity and expression of apoptosis-related proteins and SphK1/S1P/S1PR1 pathway-related proteins were, respectively, analyzed by MPO ELISA kit and Western blot analysis. The cell apoptosis in lung tissues was observed by TUNEL assay. Acute ethanol intoxication (EtOH) decreased the survival rate of mice and exacerbated the lung injury caused by sepsis through increasing pulmonary vascular permeability, neutrophil infiltration, release of inflammatory factors, and cell apoptosis. In addition, EtOH could activate the SphK1/S1P/S1PR1 pathway in CLP mice. However, PF-543, as a specific inhibitor of SphK1, could partially reverse the deleterious effects on lung injury of CLP mice. PF-543 alleviated lung injury caused by sepsis in acute ethanol intoxication rats by suppressing the SphK1/S1P/S1PR1 signaling pathway.

急性乙醇中毒会增加败血症的风险并加重败血症和肺损伤的症状。因此，本研究旨在探讨鞘氨醇激酶1（SphK1）/1-磷酸鞘氨醇（S1P）/S1P受体1（S1PR1）信号通路在急性乙醇中毒加重脓毒症肺损伤中的作用及其潜在机制。机制。采用32%乙醇溶液腹腔注射小鼠模拟急性乙醇中毒模型，采用盲肠结扎穿刺法(CLP)构建脓毒症模型。用苏木精-伊红（H&E）染色观察肺组织损伤情况，用干湿比（W/D）评价肺水肿程度。支气管肺泡灌洗液 (BALF) 中的炎症细胞计数和蛋白质浓度分别为 采用血细胞计数器和二辛可宁酸（BCA）法检测。通过其商业酶联免疫吸附测定（ELISA）试剂盒检测BALF中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-6、IL-1β和IL-18的水平。分别采用MPO ELISA试剂盒和Western blot分析髓过氧化物酶（MPO）活性和凋亡相关蛋白和SphK1/S1P/S1PR1通路相关蛋白的表达。TUNEL法观察肺组织细胞凋亡。急性乙醇中毒（EtOH）通过增加肺血管通透性、中性粒细胞浸润、炎症因子释放和细胞凋亡，降低了小鼠的存活率，并加剧了脓毒症引起的肺损伤。此外，EtOH 可以激活 CLP 小鼠的 SphK1/S1P/S1PR1 通路。然而，PF-543，作为 SphK1 的特异性抑制剂，可以部分逆转对 CLP 小鼠肺损伤的有害影响。PF-543 通过抑制 SphK1/S1P/S1PR1 信号通路减轻急性乙醇中毒大鼠脓毒症引起的肺损伤。