基于大鼠体内代谢的壮药三七姜抗炎药效物质基础研究

Study on pharmacodynamic substances of anti-inflammatory effect of Zhuang medicine Stahlianthus involucratus based on metabolism in rats

目的 为壮药三七姜的药效物质基础研究提供参考。方法 将24只雄性SD大鼠随机分为空白组(纯化水)、给药组(三七姜醇提物,以生药量计为15.75 g/kg),每组12只。每天灌胃药液/纯化水2次,每次灌胃间隔6~8 h,连续3 d。给药结束后,收集2组大鼠血液、尿液及粪便样品。利用超高效液相色谱联用四极杆-静电场轨道阱高分辨质谱技术对三七姜醇提物中的化学成分及大鼠灌胃三七姜醇提物后的血清、尿液、粪便代谢物进行鉴定。采用多元统计分析方法筛选血清中差异代谢物,并使用MetaboAnalyst 5.0平台对代谢通路进行分析。结果 从三七姜醇提物中鉴定出了38个化合物,从尿液样品中鉴定出了14个原型成分和3个代谢产物,从粪便样品中鉴定出了9个原型成分,从血清样品中鉴定出了10个原型成分和1个代谢产物。在2组大鼠血清样本中共筛选得到71个差异代谢物,其中给药组中阿魏酸、甘草素等44个差异代谢物的水平相较于空白组上调,花生四烯酸、苯乙酰谷氨酰胺等27个差异代谢物的水平相较于空白组下调。71个差异代谢物主要富集在苯丙氨酸代谢、亚油酸代谢、花生四烯酸代谢、色氨酸代谢等11条代谢通路上。结论 阿魏酸、甘草素、异嗪皮啶和刺芒柄花素等成分可能是三七姜直接发挥抗炎药效作用的物质基础,三七姜可能通过影响花生四烯酸代谢和色氨酸代谢等通路发挥抗炎作用。

OBJECTIVE To provide reference for basic analysis of the pharmacodynamic substance in Stahlianthus involucratus.METHODS Overall 24 SD male rats were randomly divided into blank group(purified water),and administration group(ethanol extract of S.involucratus,15.75 g/kg,calculated by crude drug),with 12 rats in each group.They were given drug liquid/purified water intragastrically,twice a day,every 6-8 h,for consecutive 3 days.After medication,the blood,urine and fecal samples were collected from two groups of rats.UPLC-Q-Exactive-MS technology was used to identify the chemical constituents in the ethanol extract of S.involucratus,and metabolites in the blood,urine and fecal of rats after intragastrical administration of the ethanol extract of S.involucratus.Multivariate statistical analysis was employed to screen various serum metabolites.Metabolic pathways were analyzed by MetaboAnalyst 5.0 platform.RESULTS A total of 38 chemical constituents were identified from the ethanol extract of S.involucratus,including fourteen prototype components and three metabolites identified from urine samples,nine prototype components identified from fecal samples,and ten prototype components and one metabolite identified from serum samples.A total of 71 differential metabolites were screened from two groups of rat serum samples,of which 44 differential metabolites,such as ferulic acid,glycyrrhizin,were up-regulated and 27 differential metabolites,such as arachidonic acid,phenylacetylglutamine,were down-regulated.The 71 differential metabolites were mainly enriched in 11 metabolic pathways,including phenylalanine metabolism,linoleic acid metabolism,arachidonic acid metabolism,and tryptophan metabolism.CONCLUSIONS Ferulic acid,liquiritigenin,isofraxidin and formononetin may be the material basis that directly exert pharmacological effects of S.involucratus.S.involucratus may exert anti-inflammatory effects by affecting metabolic pathways,including arachidonic acid metabolism and tryptophan metabolism.