目的 探究E1A激活基因阻遏子（CREG）调节PTEN诱导激酶1（PINK1）/帕金森病少年型蛋白2（Parkin）促进线粒体自噬对脓毒症诱导急性肺损伤（sepsis-induced acute lung injury，S-ALI）的作用。 方法 肺泡巨噬细胞株MH-S经过1、5、10及15 μg/mL脂多糖（LPS）培养后采用细胞计数试剂盒-8（CCK-8）检测MH-S细胞活性，免疫印迹及定量逆转录聚合酶链式反应（qRT-PCR）检测CREG蛋白及mRNA表达。经5 μg/mL LPS培养后细胞活性最高，采用此浓度的LPS进行后续实验。MH-S细胞分为正常组、LPS组、LPS + pLNCX2-CREG组及LPS + pSM2-siCREG组。除正常组外其余各组均经LPS培养，LPS + pLNCX2-CREG组、LPS + pSM2-siCREG组均经5 μg/mL LPS培养后分别转染质粒pLNCX2-CREG质粒及pSM2-siCREG质粒，正常组、LPS组不转染。溶酶体红色荧光探针（Lyso-Tracker Red）实验检测细胞溶酶体；流式细胞仪检测分化簇（CD）86及CD206表达；酶联免疫吸附（ELISA）试剂盒检测各组细胞白细胞介素（IL）-1β、IL-6、IL-10、C反应蛋白（CRP）、活性氧簇（ROS）、丙二醛（MDA）、超氧化物歧化酶（SOD）水平；免疫印迹及qRT-PCR检测CREG、PINK1、Parkin及蛋白及mRNA表达。 结果 与1 μg/mL LPS处理的相比，LPS 5 μg/mL的MH-S细胞活性升高（P < 0.05）；与LPS 5 μg/mL相比，LPS 10 μg/mL、LPS 15 μg/mL MH-S细胞活性均降低，且LPS 15 μg/mL细胞活性最低（P < 0.05）；与LPS 1 μg/mL相比，LPS 5 μg/mL的CREG蛋白及mRNA均差异有统计学意义（P < 0.05）；与LPS 5 μg/mL相比，LPS 10 μg/mL、LPS 15 μg/mL的CREG蛋白及mRNA均差异有统计学意义（P < 0.05）；与正常组相比，LPS组溶酶体数量、CD206、SOD、CREG、Parkin、PINK1均降低，CD86、IL-1β、IL-6、IL-10、CRP、ROS、MDA均增加（P < 0.05），与LPS组相比，LPS + pLNCX2-CREG组溶酶体数量、CD206、IL-10、SOD、CREG、Parkin、PINK1均升高，CD86、IL-1β、IL-6、CRP、ROS、MDA均降低（P < 0.05）；与LPS + pLNCX2-CREG组相比，LPS + pSM2-siCREG组酶体数量溶酶体数量、CD206、IL-10、SOD、CREG、Parkin、PINK1均降低，CD86、IL-1β、IL-6、CRP、ROS、MDA均升高（P < 0.05）。 结论 过表达CREG可增强溶酶体活性，发挥抗炎、抗氧化作用，认为CREG通过激活PINK1/Parkin通路促进线粒体自噬，从而减轻LPS诱导的S-ALI。

Objective To investigate the role of cellular repressor of E1A-stimulated genes （CREG） in alleviating lipopolysaccharide （LPS）-induced sepsis-induced acute lung injury（S-ALI） by regulating the PTEN-induced kinase 1 （PINK1）/Parkin pathway to promote mitophagy. Methods Alveolar macrophage cell line MH-S was treated with LPS at concentrations of 1， 5， 10， and 15 μg/mL. Cell viability was detected using the Cell Counting Kit-8 （CCK-8） assay， while CREG protein and mRNA expressions were measured via Western blotting and quantitative reverse transcription-polymerase chain reaction （qRT-PCR）. The 5 μg/ml LPS concentration was selected for subsequent experiments due to its ability to induce the highest cell viability. MH-S cells were divided into four groups： Normal group， LPS group， LPS + pLNCX2-CREG group， and LPS + pSM2-siCREG group. Except for the Normal group， all other groups were exposed to 5 μg/mL LPS. The LPS + pLNCX2-CREG group and LPS + pSM2-siCREG group were transfected with pLNCX2-CREG plasmid and pSM2-siCREG plasmid， respectively， after LPS treatment， while the Normal and LPS groups received no transfection. Lysosomal activity was assessed using the Lyso-Tracker Red fluorescent probe. The expressions of cluster of differentiation （CD） 86 and CD206 were detected by flow cytometry. Enzyme-linked immunosorbent assay （ELISA） kits were used to measure the levels of interleukin （IL）-1β， IL-6， IL-10， C-reactive protein （CRP）， reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） in each group. Western blot and qRT-PCR were performed to determine the protein and mRNA expressions of CREG， PINK1， and Parkin. Results Compared with the 1 μg/mL LPS group， the MH-S cell viability in the 5 μg/mL LPS group was significantly increased （P < 0.05）. In contrast， cell viability was remarkably decreased in the 10 μg/mL and 15 μg/mL LPS groups compared with the 5 μg/mL LPS group， with the lowest viability observed in the 15 μg/mL LPS group （P < 0.05）. The protein and mRNA expressions of CREG were significantly higher in the 5 μg/mL LPS group than in the 1 μg/mL LPS group （P< 0.05）， but were notably reduced in the 10 μg/mL and 15 μg/mL LPS groups compared with the 5 μg/mL LPS group （P < 0.05）. Compared with the Normal group， the LPS group exhibited decreased lysosomal quantity， CD206 expression， SOD activity， and the protein/mRNA expressions of CREG， PINK1， and Parkin， along with increased CD86 expression， levels of IL-1β， IL-6， IL-10， CRP， ROS， and MDA （all P < 0.05）. Compared with the LPS group， the LPS + pLNCX2-CREG group showed significantly elevated lysosomal quantity， CD206 expression， IL-10 level， SOD activity， and the protein/mRNA expressions of CREG， PINK1， and Parkin， as well as reduced CD86 expression and levels of IL-1β， IL-6， CRP， ROS， and MDA （all P<0.05）. Conversely， compared with the LPS + pLNCX2-CREG group， the LPS + pSM2-siCREG group displayed decreased lysosomal quantity， CD206 expression， IL-10 level， SOD activity， and the protein/mRNA expressions of CREG， PINK1， and Parkin， along with increased CD86 expression and levels of IL-1β， IL-6， CRP， ROS， and MDA （all P < 0.05）. Conclusion Overexpression of CREG enhances lysosomal activity and exerts anti-inflammatory and antioxidant effects. These findings suggest that CREG alleviates LPS-induced sepsis-induced lung injury by activating the PINK1/Parkin pathway to promote mitophagy.