桃叶珊瑚苷对UVA致ESF-1细胞光老化保护作用的代谢组学研究

摘要

Objective To analyze the mechanism and effective target of aucubin intervene photoaging ESF-1 cells by high through-put cell metabolomics.Methods Cell photoaging model was established by UVA cell injury method,then given low medium and high dose aucubin DMEM medium for the intervention.The activities and contents of total superoxide dismutase (T-SOD),glutathione peroxidase (GSH-Px),catalase (CAT),malondialdehyde (MDA) were detected for the evaluation of oxidative damage and aucubin treatment effect.Based on above analysis and high resolution mass spectrometry,the high throughput metabolic footprint analysis was conducted in the normal cells,photoaging ESF-1 cells and aucubin intervention cell.Massive metabolic data were analzyed with dinension reduction process and multivariate statistics by model discriminating technique.Results Compared with the blank group,the activities of SOD,GSH-Px,CAT significantly decreased and contents of MDA significantly increased in the model group (P ＜ 0.01).Compared with the model group,the activities of SOD,GSH-Px,CAT increased and contents of MDA decreased in the medication gorups,of which,in the medium and high doses medication groups the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01).There were significant differences in the metabolism between the normal cells and the photoaging ESF-1 cells,and the disturbance of metabolism was mainly concentrated in arachidonic acid metabolism,glutathione metabolism.6 core biomarkers were identified by focusing analysis,and these small molecular metabolites were callback after the administration of aucubin.Conclusion Aucubin has a favorable effect on photoaging,and the mechanism may be related to arachidonic acid metabolism.Besides,the present investigation provides new ideas and methods for early prevention and drug development.%目的 采用高通量细胞代谢组学技术分析桃叶珊瑚苷干预光老化ESF-1细胞的效应机制和有效作用靶点.方法 细胞UVA损伤模型法制备光老化细胞模型,给予低中高不同剂量的桃叶珊瑚苷DMEM培养液进行干预,通过检测细胞所含总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化酶(GSH-Px)、过氧化氢酶(CAT)、丙二醛(MDA)的活性和含量评价光老化ESF-1细胞模型的氧化损伤情况及桃叶珊瑚苷对模型的干预作用.在此基础上基于高分辨质谱技术对正常细胞、光老化细胞及桃叶珊瑚苷干预后的细胞进行高通量的代谢足迹分析.通过模式识别技术对海量的数据采取降维处理和多元统计分析.结果 与空白组相比,模型组细胞中SOD、GSH-Px、CAT活性明显降低,MDA含量明显升高(P＜0.01);与模型组相比,给药组能提高SOD、GSH-Px、CAT活性,降低MDA含量,其中中、高剂量组差异有统计学意义(P＜ 0.05或P＜ 0.01).光老化细胞代谢与空白组细胞存在显著性差异,发生扰乱的代谢主要集中在花生四烯酸代谢、谷胱甘肽代谢.通过聚焦分析发现了6个核心的生物标志物,且这些小分子代谢产物在给予桃叶珊瑚苷干预后都发生了有效的回调.结论 桃叶珊瑚苷对光老化有很好的治疗作用,其作用机制可能与花生四烯酸代谢机制相关,本研究为早期预防和药物研发提供了新思路和新方法.