基于代谢组学研究蚕沙对湿阻中焦证大鼠的“和胃”作用机制

Gastric-harmonizing mechanisms of Cansha in rats with Shizhuzhongjiao syndrome based upon metabolomics

目的:研究蚕沙对湿阻中焦证大鼠粪便及尿液中内源性代谢物的影响,探讨蚕沙治疗湿阻中焦证可能机制。方法:利用"外湿侵体法-游泳力竭法-过食肥甘法"多因素复合法制备湿阻中焦证大鼠模型;采用超高效液相色谱-飞行时间质谱(UPLC-Q-TOF-MS)技术结合多变量统计分析方法,研究湿阻中焦证大鼠粪便和尿液代谢物谱图的变化,确定相关代谢标志物并构建代谢通路。结果:与正常组比较,模型组大鼠粪便及尿液中十八碳-6,9-二烯酸、油酸酰胺、十八碳6烯酸、14,15-二羟基二十碳三烯酸、1-(1Z-十八碳烯基)-sn-甘油、二十四碳五烯酸(C24:5n-6)、7-酮基脱氧胆酸、α-生育三烯酚、溶血磷脂酰乙醇胺(0∶0/15∶0)、溶血磷脂酰乙醇胺(0∶0/18∶1(11Z))、溶血磷脂酰胆碱(18∶3(6Z,9Z,12Z))、半胱氨酸、硫酸吲哚酚、蓖麻油酸、环磷酸鸟苷显著升高;3-吲哚甲醛、4,6-二羟基喹啉、L-苯丙氨酸、犬尿喹啉酸、α-亚麻酸、甘氨胆酸、2-吲哚羧酸、乙酰磷酸、高香草酸、黄尿酸、棕榈酸、亚油酸、油酸显著降低。同时,蚕沙高、中、低剂量组分别能显著回调其中的17、13、11种代谢物。结论:代谢组学分析揭示蚕沙对湿阻中焦证大鼠具有一定的干预作用,其"和胃"机制可能与调节苯丙氨酸、酪氨酸和色氨酸的生物合成,苯丙氨酸代谢,α-亚麻酸代谢,甘油磷脂代谢,初级胆汁酸的生物合成等有关。

OBJECTIVE To explore the effects of Cansha(Faeces Bombycis)upon endogenous metabolites in rats with Shizhuzhongjiao syndrome to clarify the underlying mechanisms of Cansha on Shizhuzhongjiao syndrome.METHODS The model of Shizhuzhongjiao syndrome was developed by complex factors including an invasion of external dampness,exhaustive swimming and overfeeding of high fat and carbohydrate diets.Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)and multivariate statistical analysis were utilized for examining the changes of metabolites in feces and urine of rats with Shizhuzhongjiao syndrome and identifying biomarkers and metabolic pathways related to Shizhuzhongjiao syndrome.RESULTS As compared with control group,the fecal and urinary levels of 15 metabolites including octadecanoic-6,9-dienoic acid,oleamide,octadeca-6-enoic acid,14,15-dihydroxyeicosatentaenoic acid,1-(1Z-octadecenyl)-Sn-glycerin,Tetracosapentaenoic acid(C24:5n-6),7-ketodeoxycholic acid,α-tocotrienol,lysophosphatidylethanolamine(0∶0/15∶0),lysophosphatidylethanolamine(0∶0/18∶1(11Z)),lysophosphatidylcholine(18∶3(6Z,9Z,12Z)),cysteine,indole phenol sulfate,ricinoleic acid and guanosine cyclophosphate became elevated in model group.The fecal and urinary levels of 13 metabolites including 3-indole-formaldehyde,4,6-dihydroxyquinoline,L-phenylalanine,caninuria quinolinic acid,α-linolenic acid,glycocholic acid,2-indole carboxylic acid,acetyl phosphoric acid,homovanillic acid,xanthic acid,palmitic acid,linoleic acid and oleic acid declined in model group.And the levels of 17,13 and 11 metabolites were restored to normal levels after offering high/medium/low-dose of Cansha respectively.CONCLUSION Metabolomic analysis revealed that Cansha had certain therapeutic effects on rats with Shizhuzhongjiao syndrome.And Hewei mechanism may be correlated with the biosynthesis of phenylalanine,tyrosine and tryptophan,phenylalanine metabolism,α-linolenic acid metabolism,glycerin phospholipid metabolism and primary bile acid biosynthesis.