基于非靶向代谢组学探索辐射诱导的小鼠肺上皮细胞代谢改变

Non-targeted Metabolomics-based Exploration of Radiation-induced Metabolic Alterations in Mouse Lung Epithelial Cells

背景与目的代谢变化是放射性肺炎的重要特征之一。放疗作为胸部肿瘤治疗的常规手段,在有效杀伤肿瘤细胞的同时也会引发局部炎症和纤维化等不良反应,进而导致治疗效果受限,严重影响患者生活质量。因此,深入探讨放疗引起的代谢改变具有重要意义。本研究旨在探究X射线辐射对小鼠肺上皮细胞系(murine lung epithelial-12,MLE12)代谢的影响。方法于体外环境下培养MLE12细胞系,并将其随机分为辐射组(radiation group,IR组)与对照组(control group,NC组)。使用日立X射线辐照仪以10 Gy的剂量对IR组细胞进行辐射处理。辐射后48 h收集细胞上清液样本。通过液相色谱-质谱联用仪(liquid chromatograph mass spectrometer,LC/MS)对样本进行代谢组学分析。结果LC/MS代谢组学分析揭示了MLE12细胞在辐射后48 h的代谢变化。与NC组相比,IR组中共有38种分泌型代谢物发生改变。根据京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库的注释,差异代谢物主要涉及核苷酸代谢、氨基酸代谢和脂质代谢等通路,其中核苷酸代谢的差异最为显著。结论X射线辐射对MLE12细胞的代谢产生了显著影响,主要影响核苷酸代谢途径,包括嘌呤和嘧啶等代谢产物及相关代谢通路。

Background and objective Metabolic change is one of the important characteristics of radiation pneumonitis.Radiotherapy,as a conventional method for the treatment of thoracic tumors,can not only effectively kill tumor cells,but also cause adverse reactions such as local inflammation and fibrosis,which leads to limited therapeutic effect and profound impact on the quality of life of patients.Therefore,it is of great significance to explore the metabolic changes caused by radiotherapy.The aim of this study was to investigate the effects of X-ray irradiation on the metabolism of a mouse lung epithelial cell line(murine lung epithelial-12,MLE12).Methods MLE12 cells were cultured in vitro and randomly divided into radiation group(IR)and control group(NC).Cells in the IR group were irradiated at a dose of 10 Gy using a Hitachi X-ray irradiator.Cell supernatant samples were collected at 48 h after irradiation.Metabolomic analysis of the samples was performed by liquid chromatograph mass spectrometer(LC/MS).Results LC/MS metabolomics analysis revealed the metabolic changes of MLE12 cells at 48 h after irradiation.A total of 38 secretory metabolites were altered in the IR group compared with the NC group.According to the annotation of Kyoto Encyclopedia of Genes and Genomes(KEGG)database,the differential metabolites are mainly involved in nucleotide metabolism,amino acid metabolism and lipid metabolism,among which the difference in nucleotide metabolism is the most significant.Conclusion The metabolism of MLE12 cells was significantly affected by X-ray irradiation,mainly affecting the nucleotide metabolic pathways,including purine and pyrimidine metabolites and related metabolic pathways.