基于UPLC-QE-Orbitrap-MS的生姜干预腹泻小鼠血清代谢组学分析  

Serum Metabonomics Analysis of Diarrhea Mice Intervened by Zingiberis Rhizoma Recens Based on UPLC-QE-Orbitrap-MS

作　　者：玄静[1] 刘燕[1] 张永清[1] XUAN Jing;LIU Yan;ZHANG Yong-qing(Key Laboratory of Chinese Medicine Resources in Colleges and Universities of Shandong Province,Shandong University of Traditional Chinese Medicine,Jinan 250355,China)

机构地区：[1]山东中医药大学山东省高等学校中药资源学重点实验室,济南250355

出　　处：《中国实验方剂学杂志》2022年第1期157-164,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山东省重点研发计划项目(2016CYJS08A01-2);2018年中医药公共卫生服务补助专项(财社[2018]43号);2019年医疗服务与保障能力提升项目(财社[2019]39号)。

摘　　要：目的:探讨生姜对腹泻小鼠的止泻作用及其机制,为生姜抑制肠蠕动及其用于胃肠疾病的治疗提供研究基础。方法:采用番泻叶建立小鼠腹泻模型,设定正常组、模型组、生姜低、中、高剂量(0.1,0.32,1.0 g·kg^(-1))组和洛哌丁胺组(给药剂量1.6 g·kg^(-1)),通过小鼠腹泻评分,稀便发生率(LSIR),平均稀便等级(ALSG),腹泻指数(DI),肠推进率,肠组织病理切片检测生姜不同剂量组对腹泻小鼠的干预作用;利用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法(UPLC-QE-Orbitrap-MS),主成分分析(PCA)及正交偏最小二乘法-判别分析(OPLS-DA)对小鼠血清进行代谢组学分析,流动相选择0.1%甲酸水溶液(A)-0.1%甲酸乙腈溶液(B)梯度洗脱(0~3.5 min,5%~15%B;3.5~6 min,15%~30%B;6~6.5 min,30%B;6.5~12 min,30%~70%B;12~12.5 min,70%B;12.5~18 min,70%~100%B),流速0.4 mL·min^(-1),进样量5μL;电喷雾离子源(ESI),正、负离子检测模式,采集范围m/z 100~1 500。结果:与模型组比较,生姜高剂量组可明显降低腹泻评分,LSIR,ALSG,DI和肠推进率(P<0.05,P<0.01),改善肠黏膜损伤;正常组、模型组和生姜高剂量组3组之间的主要差异代谢物共有40种,包括葡萄糖1-磷酸、黄嘌呤、黄嘌呤核苷和酵母甾醇中间体2等;涉及的代谢通路主要有淀粉和蔗糖代谢、氨基糖和核苷酸糖代谢、果糖和甘露糖代谢、半乳糖代谢、色氨酸代谢等。结论:生姜可抑制腹泻小鼠肠蠕动进而发挥止泻作用,作用机制与调节糖代谢、氨基酸代谢有关。Objective: To investigate the antidiarrheal effect and mechanism of Zingiberis Rhizoma Recens on diarrhea mice,and to provide research basis for the inhibition of intestinal peristalsis by Zingiberis Rhizoma Recens and its application in the treatment of gastrointestinal diseases. Method:The diarrhea model of mice was established by Sennae Folium. The control group,model group,Zingiberis Rhizoma Recens low-,medium-, high-dose groups(0.1, 0.32, 1.0 g·kg^(-1)) and loperamide group(1.6 g·kg^(-1)) were set. The intervention effect of Zingiberis Rhizoma Recens with different doses on diarrhea mice was detected by diarrhea score,incidence rate of loose stools(LSIR),grade of average loose stools(ALSG),diarrhea index(DI),intestinal propulsion rate and intestinal pathological section. The serum metabonomics of mice was analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-QE-Orbitrap-MS), principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). The conditions were as follows:mobile phase of 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B)for gradient elution(0-3.5 min,5%-15%B;3.5-6 min,15%-30%B;6-6.5 min,30%B;6.5-12 min,30%-70%B;12-12.5 min,70%B;12.5-18 min,70%-100%B),flow rate of 0.4 mL·min^(-1),injection volume of 5 μL,electrospray ionization(ESI),positive and negative ion detection modes,acquisition range of m/z 100-1 500. Result: Compared with the model group,Zingiberis Rhizoma Recens high-dose group could obviously reduce the diarrhea score,LSIR,ALSG,DI and intestinal propulsion rate(P<0.05,P<0.01),and improve the intestinal mucosal injury. There were 40 main differential metabolites among the control group,model group and Zingiberis Rhizoma Recens high-dose group,including glucose 1-phosphate,xanthine,xanthosine and so on. The metabolic pathways mainly included starch and sucrose metabolism,amino sugar and nucleotide sugar metabolism,fructose and mannose metabol

关 键 词：生姜 腹泻 代谢组学 差异代谢物 代谢通路 作用机制 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法(UPLC-QE-Orbitrap-MS) 

分 类 号：R22[医药卫生—中医基础理论] R28[医药卫生—中医学]