黄芩-栀子配伍治疗糖尿病肾病大鼠的尿液代谢组学研究

采用基于超高效液相色谱与串联四极杆飞行时间质谱仪(Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,UHPLC-Q-TOF MS)联用的代谢组学方法,通过分析大鼠尿液内源性代谢物的变化,研究黄芩-栀子药对治疗糖尿病肾病的作用机制。利用高脂高糖饲料喂养并腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型。给药12周后,采用UHPLC-Q-TOF MS对正常对照组、模型组、黄芩-栀子药对治疗组的尿液样品进行代谢轮廓分析。利用正交偏最小二乘判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)对实验数据进行分析,挑选出对各组分分离贡献较大(VIP>1,p<0.05)的化合物在Human Metabolome Database(HMDB)等数据库进行质谱信息匹配,共鉴定出糖尿病肾病潜在生物标记物31个,给药后回调生物标记物16个。研究结果表明,黄芩-栀子药对通过影响胆汁酸的合成与代谢、尿毒素代谢、能量代谢、色氨酸代谢、苯丙氨酸代谢、酪氨酸代谢等通路对糖尿病肾病起到治疗作用,其中胆汁酸的合成与代谢、尿毒素代谢和能量代谢可能是黄芩-栀子药对发挥治疗作用的主要途径。与单味药相比,配伍后的药对具有了新的药理作用。

Urinary Metabolomics Study of Rat Suffering Diabetic Nephropathy Treated with Radix Scutellariae-Gardenia jasminoides Couples

The volatile oils of Radix Bupleuri extracted under decompression and atmospheric pressure were compared in terms of composition,content and efficacy to investigate the quality of the volatile oil and optimize the extraction method.Gas chromatography-mass spectrometry(GC-MS) was used to analyze the constituents of volatile oils and their relative contents.Meanwhile,a method of gas chromatography(GC) was established for the determination of hexanal,heptanal,octanal and trans 2,4 decadienal in the volatile oils,and the contents of active components in volatile oil extracted under different pressures were determined.A rat fever model was reproduced by subcutaneous injection of yeast suspension on the back,the volatile oils extracted from Radix Bupleuri under atmospheric pressure and decompression were dissolved in 1% Tween 80 and injected intraperitoneally.The temperature changes of rats at different time points were measured,and single factor analysis of variance was used for intergroup comparison.The composition and content of the volatile oil extracted under decompression conditions were different from those extracted under atmospheric pressure,the extraction rates of hexanal,heptanal,octanal and trans 2,4 decadienal were higher,and the effect of the volatile oil on the fever caused by dry yeast was more significant.Results showed that decompression could effectively reduce the damage of heat sensitive components and improve the extraction efficiency of effective components.The volatile oil of Radix Bupleuri extracted by this method has a more significant medicinal effect,and its quality is better than that extracted under atmospheric pressure.