基于UPLC-Q-Exactive MS技术的宣白承气汤加味治疗流感病毒与肺炎链球菌共感染小鼠肺炎代谢组学研究

Metabolomics Study of Modified Xuanbai Chengqi Decoction(宣白承气汤加味)in Treatment of Pneumonia in Influenza Virus/Streptococcus Pneumoniae Co-Infected Mice Based on UPLC-Q-Exactive MS Technology

目的基于超高效液相-四级杆-轨道阱高分辨质谱联用(Ultrahigh-performance liquid chromatography-quadrupole exactive mass spectrometry, UPLC-Q-Exactive MS)技术探究宣白承气汤加味对流感病毒/肺炎链球菌(Influenza virus/Streptococcus pneumoniae,Ⅳ/Spn)共感染小鼠肺脏组织中内源性代谢产物的影响,旨在阐明宣白承气汤加味治疗共感染小鼠肺炎的作用机制。方法将36只小鼠随机分为3组,即正常组、模型组和宣白承气汤加味组,每组12只。除正常组外,其余两组小鼠在造模成功6 h后开始灌胃给药,1次/d,连续1周。给药结束后,取小鼠肺脏组织,利用UPLC-Q-Exactive MS技术进行非靶向代谢组学研究,结合主成分分析(Principal component analysis, PCA)和正交偏最小二乘判别分析法(Orthogonal partial least-squares discrimination analysis, OPLS-DA)分析并筛选潜在的代谢产物,并基于R语言对已鉴定的差异性代谢产物进行通路分析。结果在正负离子模式下,共筛选和鉴定出36个潜在的代谢产物,与正常组相比,模型组有27个代谢产物含量显著上升,有9个代谢产物含量显著下降。宣白承气汤加味干预后,36个代谢产物的含量均呈现明显的回调趋势,主要影响亚油酸代谢、烟酸和烟酰胺代谢、核黄素代谢等通路。结论宣白承气汤加味对Ⅳ/Spn共感染小鼠肺炎的治疗作用可能与其调节体内代谢产物失调,恢复体内正常生命活动有关。

Objective To investigate the effect of Modified Xuanbai Chengqi Decoction(宣白承气汤加味)on endogenous metabolites in lung tissues of influenza virus/streptococcus pneumoniae(IV/Spn)co-infected mice based on ultrahigh-performance liquid chromatography-quadrupole exactive mass spectrometry(UPLC-Q-Exactive MS),aiming to elucidate the mechanism of action of Modified Xuanbai Chengqi Decoction in the treatment of pneumonia in co-infected mice.Methods Thirty-six mice were randomly divided into 3 groups,namely normal group,model group and Modified Xuanbai Chengqi Decoction group,with 12 mice in each group.Except for the normal group,the mice of the remaining two groups started administration by gavage once a day for one week after 6 h of successful modeling.After the end of administration,the lung tissue was taken out and untargeted metabolomics studies were carried out by using UPLC-Q-Exactive MS technology,combined with principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to analyze and screen potential metabolites,and pathway analysis of identified differential metabolites was carried out based on R software.Results A total of 36 potential metabolites were screened and identified in positive and negative ion mode,and compared with the normal group,the contents of 27 metabolites in the model group increased significantly,and the contents of 9 metabolites decreased significantly.After the intervention of Modified Xuanbai Chengqi Decoction,the contents of 36 metabolites showed a clear correction trend,mainly affecting linoleic acid metabolism,niacin and nicotinamide metabolism,riboflavin metabolism and other pathways.Conclusion The therapeutic effect of Modified Xuanbai Chengqi Decoction on pneumonia in IV/Spn co-infected mice may be related to its regulation of metabolic imbalance in vivo and the restoration of normal life activities in vivo.