基于UHPLC-Q-Exactive尿液代谢组学的丹贝益肺方对肺纤维化干预作用研究

Intervention effect of Danbei Yifei formula on pulmonary fibrosis based on urine metabolomics by UHPLC-Q-Exactive

目的:确定肺纤维化大鼠的尿液生物标记物,阐述丹贝益肺方干预肺纤维化的作用机制。方法:采用气管雾化博莱霉素方法诱导肺纤维化大鼠模型,测定临床肺纤维化的诊断指标,如超氧化物歧化酶、谷胱甘肽、丙二醛。通过最新高分辨率液质联用技术,非靶向获取肺纤维化大鼠的高通量代谢组数据,并以化学计量学算法对多维数据进行数据处理,筛选与肺纤维化相关的生物标记物。同时,观察给药后大鼠的各项代谢机能指标,阐述丹贝益肺方对肺纤维化的有效作用机制。结果:临床生化指标显示,模型组大鼠代谢和健康大鼠相比差异显著,证实肺纤维化大鼠模型制备成功。代谢组学研究表明,肺纤维化大鼠代谢轮廓明显偏移,体内代谢异常;给予丹贝益肺方后整体呈回调趋势,并向健康大鼠代谢轮廓方向发展。经数据库匹配和数据处理后,得到健康大鼠与肺纤维化大鼠组间显著变化的12个生物标记物,包括富马酸盐、精氨酸、亚精胺等。结论:丹贝益肺方对肺纤维化大鼠有一定的治疗作用。调节三羧酸循环及精氨酸代谢通路等可能是其治疗肺纤维化的作用机制。

Objective:To determine the urine biomarkers of rats with pulmonary fibrosis and to elucidate the intervention mechanism of Danbei Yifei formula.Methods:Bleomycin was injected into trachea to induce pulmonary fibrosis in rats.High throughput metabolomics data of rats with pulmonary fibrosis were obtained by the latest high-resolution liquid chromatography mass spectrometry,and the multi-dimensional data were processed by chemometrics algorithm to screen biomarkers related to pulmonary fibrosis.Meanwhile,the metabolic function indexes of rats after administration were observed,and the effective mechanism of Danbei Yifei formula on pulmonary fibrosis was elaborated.Results:The clinical biochemical indexes showed that there were significant differences in the metabolism of rats in the model group,which confirmed that the rat model of pulmonary fibrosis was successfully prepared.Metabolomics study showed that the metabolic profile of pulmonary fibrosis rats was obviously deviated,and the metabolism was abnormal in vivo.After Danbei Yifei formula was given,it showed a reverse trend and developed towards normal level.After database matching and data processing,12 biomarkers with significant changes between the healthy rats and the rats with pulmonary fibrosis were obtained,namely fumarate,arginine and spermidine,etc.Conclusion:Danbei Yifei formula has a certain therapeutic effect on pulmonary fibrosis rats.Regulating the tricarboxylic acid cycle and arginine metabolism pathway may be its mechanism in treating pulmonary fibrosis.