基于UHPLC-Q-Exactive细胞代谢组学技术探讨丹贝益肺方对人胚肺成纤维细胞的作用机制  

Study of Mechanism of Danbei Yifei Decoction(丹贝益肺方)on Human Embryonic Lung Fibroblasts Based on UHPLC-Q-Exactive Cell Metabolomics Technology

作　　者：蔡萧君 李宇 王钦 吴圆圆 胡杨 颉彦鹏 CAI Xiaojun;LI Yu;WANG Qin;WU Yuanyuan;HU Yang;XIE Yanpeng(Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150036,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036

出　　处：《山东中医药大学学报》2022年第6期738-746,765,共10页Journal of Shandong University of Traditional Chinese Medicine

基　　金：黑龙江省应用技术研究与开发计划项目(编号:GA19C108)。

摘　　要：目的:基于代谢组学技术探讨丹贝益肺方对人胚肺成纤维细胞(MRC-5)的作用机制。方法:将经丹贝益肺方提取物预处理24 h的MRC-5设为给药组,弃去预处理液,加入细胞维持液,在37℃、5%CO_(2)的饱和水汽二氧化碳培养箱中培养72 h后收获上清液;另取未经丹贝益肺方提取物干预的MRC-5为模型组,未经丹贝益肺方提取物干预的人胚肺正常细胞为空白组,两组其他处理与给药组一致。收集各组样本上清液,采用超高液相色谱-四极杆-静电场轨道阱高分辨质谱技术对细胞内容物进行细胞代谢组学采集,通过化学计量学算法分析MRC-5代谢数据。结果:与模型组比较,给药组细胞吸光度值升高(P<0.01),给药组细胞增殖抑制率为94%,明显高于模型组。通过Acequire X的数据依赖型扫描模式对细胞代谢产物进行采集和分析,共获得差异代谢产物6个,分别是溶血磷脂酰胆碱、磷脂酰乙醇胺、左旋肉碱、天冬氨酸、谷氨酸、谷氨酰胺,经丹贝益肺方提取物干预后全部呈现回调趋势。结论:丹贝益肺方对肺纤维化有较好的干预作用,其机制可能与调节精氨酸生物合成、天冬氨酸和谷氨酸代谢及D-谷氨酰胺代谢等有关。Objective:To investigate the mechanism of Danbei Yifei Decoction(丹贝益肺方)on human embryonic lung fibroblasts(MRC-5)based on metabolomics technology.Methods:MRC-5 pretreated with Danbei Yifei Decoction extract for 24 h was set as the administration group,the pretreatment solution was discarded,the cell maintenance solution was added,and the supernatant was harvested after culturing for 72 h in a saturated water vapor carbon dioxide incubator at 37℃and 5%CO_(2).In addition,MRC-5 without the intervention of Danbei Yifei Decoction extract was taken as the model group,and human embryonic lung normal cells without the intervention of Danbei Yifei Decoction extract were taken as the blank group.The other treatments in the two groups were consistent with the administration group.The super natant of each group of samples was collected,the cell contents were collected by ultra-high liquid chromatography quadrupole electrostatic field orbital trap high resolution mass spectrometry for cellular metabolomics,and the MRC-5 metabolic data was analyzed by chemometrics algorithm.Results:Compared with the model group,the absorbance value of cells in the administration group increased(P<0.01),and the inhibition rate of cell proliferation in the administration group was 94%,which was significantly higher than that in the model group.Cell metabolites were collected and analyzed by Acequire X′s data-dependent scanning mode,and a total of 6 differential metabolites were obtained,namely lysophosphatidylcholine,phosphatidylethanolamines,L-carnitine,aspartate acid,glutamic acid and glutamine,all showed a downward trend after intervention by Danbei Yifei Decoction extract.Conclusions:Danbei Yifei Decoction has a good intervention effect on pulmonary fibrosis,and its mechanism may be related to the regulation of arginine biosynthesis,aspartic acid and glutamic acid metabolism and D-glutamine metabolism.

关 键 词：丹贝益肺方 人胚肺成纤维细胞 肺纤维化 代谢组学 轨道阱高分辨质谱 生物信息学 

分 类 号：R285.5[医药卫生—中药学]