基于GC-TOF-MS代谢组学研究黄连解毒汤对大鼠粪便代谢的影响

Effects of Huanglian Jiedu Decoction on Fecal Metabolism in Rats Based on GC-TOF-MS Metabolomics

目的通过GC-TOF-MS代谢组学探讨不同剂量的黄连解毒汤对SD大鼠粪便代谢组的影响,进而为黄连解毒汤的作用机制提供参考。方法SD大鼠45只适应性喂养7天后,随机分为5组:黄连解毒汤高剂量组、中剂量组、低剂量组、抗生素组和空白组,每组9只。中药干预组不同剂量黄连解毒汤灌胃,1mL/次,2次/天,连续给药21天;抗生素组,1 mL/次,2次/天;空白组生理盐水灌胃,1 mL/次,2次/天,连续给药21天;末次给药后,禁食禁水12 h,处死并收集大鼠粪便。采用气相色谱-飞行时间质谱(Gas Chromatography Tandem Time-of Flight Mass Spectrometry,GC-TOF-MS)检测各组大鼠粪便,运用多元分析OPLS以及单变量统计学方法筛选差异代谢物,通过MetaboAnalyst平台进行代谢通路分析。结果对于粪便中的内源性代谢物小分子,高剂量组的黄连解毒汤影响的主要有:核糖、硫酸、棕榈酸、花生酸、茜素等;中剂量组的黄连解毒汤影响的主要有:乌头酸、胆固醇、乳酸、棕榈油酸、茜素等;低剂量组的黄连解毒汤影响的主要有:硫酸、棉子糖;氨苄西林钠影响的主要有:蛋氨酸1、麦角固醇、正亮氨酸1、棕榈酸、硫酸、乳酸等;不同剂量的黄连解毒汤和氨苄西林钠都能够影响到半乳糖代谢及硫代谢通路。结论黄连解毒汤可改善糖尿病和心脑血管疾病相关的代谢物小分子,不同剂量的黄连解毒汤对粪便代谢的影响不同,其中中剂量的影响最为广泛。

Objective To explore the effect of different doses of Huanglian Jiedu Decoction on the fecal metabolome of SD rats based on GC-TOF-MS metabolomics,and then to provide a reference for the mechanism of Huanglian Jiedu Decoction.Methods After 7 days of adaptive feeding,45 SD rats were randomly divided into 5 groups of Huanglian Jiedu Decoction high-dose group,medium-dose group,low-dose group,antibiotic group and blank group,with 9 rats in each group.In the Chinese medicine intervention group,different doses of Huanglian Jiedu Decoction were administered by gavage,1 mL per time,2 per day for 21 days;in the antibiotic group,1 mL per time,2 per day;in the blank group,saline was administered by gavage,1 mL per time,2 per day for 21 days;after the last administration,the rats were fasted with no food and water for 12 h.The rats were executed and their feces were collected.The feces of each rat group were examined by gas chromatography-time-of-flight mass spectrometry(GC-TOF-MS),and the differential metabolites were screened by multivariate analysis OPLS and univariate statistics,and the metabolic pathways were analyzed by MetaboAnalyst.The metabolic pathway analysis was performed on the MetaboAnalyst platform.Results For the endogenous metabolites in feces,the high-dose group of Huanglian Jiedu Decoction mainly affected ribose,sulfuric acid,palmitic acid,arachidic acid,alizarin,etc.;the medium dose group of Huanglian Jiedu Decoction mainly affected aconitic acid,cholesterol,lactic acid,palmitic acid,alizarin,etc.;the low-dose group of Huanglian Jiedu Decoction mainly affected sulfuric acid,raffinose,ampicillin,etc.The main effects of sodium were methionine 1,ergosterol,n-leucine 1,palmitic acid,sulfuric acid,lactic acid,etc.;different doses of Huanglian Jiedu Decoction and ampicillin sodium can affect galactose metabolism and thiometabolism pathway.Conclusion Huanglian Jiedu Decoction can improve small molecules of metabolites related to diabetes and cardiovascular and cerebrovascular diseases.Different doses of Huanglian Jiedu Decoction have different effects on fecal metabolism,and the medium dose has the most extensive effect.