基于UPLC-Q Tof MS/MS的蒙药给喜古讷-3颗粒治疗便秘大鼠尿液代谢组学研究  

Urine metabolomics of constipated rats treated with Mongolian medicine Geixigune-3 granules based on UPLC-Q Tof MS/MS

作　　者：包泽阳 孟和[2] 都丽娜 宝苏日高 韩额尔德木图 孟永梅 BAO Ze-yang;MENG He;DU Li-na;BAO Su-ri-gao;HAN E-er-de-mu-tu;MENG Yong-mei(College of Mongolian Medicine,Inner Mongolia Medical University,Hohhot 010110,China;Institute of Mongolian Medicine Chemistry,School of Chemistry and Chemical Engineering,Inner Mongolia University,Hohhot 010121,China;Inner Mongolia International Mongolian Medical Hospital,Hohhot 010013,China;Inner Mongolia Medical University Affiliated Hospital,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学蒙医药学院,呼和浩特010110 [2]内蒙古大学高分子化学与蒙药研究所,呼和浩特010021 [3]内蒙古国际蒙医医院,呼和浩特010013 [4]内蒙古医科大学附属医院,呼和浩特010050

出　　处：《药物分析杂志》2023年第7期1205-1212,共8页Chinese Journal of Pharmaceutical Analysis

基　　金：内蒙古自治区科技创新引导奖励资金项目(20170527);内蒙古医科大学附属医院一般项目(NYFYYB031);内蒙古医科大学善学计划(zy0201031);内蒙古医科大学教学名师项目(NYJXMS202203);内蒙古自治区高等学校科学研究项目(NJZZ22634)。

摘　　要：目的:观察蒙药给喜古讷-3颗粒对便秘大鼠的尿液代谢组学研究,寻找相关代谢产物及代谢通路。方法:大鼠随机分为空白组、模型组、给喜古讷-3颗粒组、阳性给药(莫沙必利)组,共4组。皮下注射盐酸洛哌丁胺建立大鼠便秘模型,观察各组大鼠24 h尿量、粪便质量、粒数、含水率,造模成功后给予相应的药物进行治疗。给药7 d后取各组大鼠尿液,利用超高效液相色谱-四极杆飞行时间串联质谱(UPLC-Q Tof MS/MS)的代谢组学技术,采用ACQUITY UPLC?BEH Shield RP 18色谱柱(100 mm×2.1 mm, 1.7μm),以0.1%甲酸水-含0.1%甲酸的乙腈为流动相进行梯度洗脱,流速0.35 mL·min^(-1),质谱采用电喷雾离子源(ESI),以正负离子模式采集质谱碎片信息,对大鼠尿液进行分析寻找差异代谢物和代谢通路。结果:与模型组比较,给喜古讷-3颗粒组24 h尿量减少(P<0.05),粪便质量增加(P<0.01),粒数增加(P<0.05),含水率升高(P<0.01)。共鉴定出差异代谢物22个,其中呋喃苯胺酸、聚酯吡喃酮D、尿苷2’,3’-环磷酸水平给药后升高,半胱氨酸-赖氨酸、去甲基多塞平、N-葡萄糖酰乙醇胺等水平给药后降低,主要涉及代谢通路为色氨酸代谢、类固醇生物合成、泛醌和其他萜类醌生物合成。结论:给喜古讷-3颗粒对便秘模型大鼠尿液代谢产物具有调节作用,为进一步对给喜古讷-3颗粒治疗便秘奠定了有效的实验基础。Objective:To observe the effect of Mongolian medicine Geixigune-3 granules on urine metabolomics in constipation rats,and search for related metabolites and metabolic pathways.Methods:Rats were randomly divided into 4 groups:blank group,model group,Geixigune-3 granules group and positive drug(mosapride)group.The constipation model of rats was established by subcutaneous injection of loperamide hydrochloride.The 24 h urine volume,fecal weight,number of fecal particals and moisture content of rats in each group were observed.After the model was successfully established,corresponding drugs were given for treatment.ACQUITY UPLC?BEH Shield RP 18(100 mm×2.1 mm,1.7μm)column with a 0.1%formic acid water and acetonitrile containing 0.1%formic acid as mobile phase were used.Gradient elution was applied and the flow rate was 0.35 mL·min^(-1).ESI with positive and negative ionspattern was used.Rat urine was analzed to identify differential metabolites and metabolic pathways.Results:Compared with the model group,the 24 h urine volume decreased(P0.05),the fecal weight increased(P<0.01),the number of particals increased(P0.05),and the moisture content increased(P<0.01)in the Geixigune-3 granules group.A total of 22 differential metabolites were identified,of which the levels of furosemide,polyporapyranone D,uridine 2',3'-cyclic phosphate increased after administration,and the levels of cysteinyl-lysine,didesmethyl doxepin and N-gluconyl ethanolamine decreased after administration,mainly involving tryptophan metabolism,ubiquinone and other terpenoid-quinone biosynthesis and steroid biosynthesis.Conclusion:Geixigune-3 granules has a regulating effect on urine metabolites of constipation model rats,which lays an effective experimental foundation for further treating constipation with Geixigune-3 granules.

关 键 词：蒙药 给喜古讷-3颗粒 便秘 大鼠 尿液 代谢组学 超高效液相色谱-四极杆飞行时间串联质谱 

分 类 号：R917[医药卫生—药物分析学]