田黄方降脂作用配伍机制的尿液代谢组学研究

Study on the compatibility rule of tianhuang fang in treating hyperlipidemia based on urine metabonomics

目的:在前期研究发现田黄方合用比单用三七、黄连对高脂血症大鼠模型血脂调节作用更优的基础上,进一步采用代谢组学技术比较其合用及单用对饮食性高脂血症大鼠尿液代谢产物的差异,揭示其治疗高脂血症的配伍机制。方法:取雄性SD大鼠30只,分为空白组、模型组、黄连组(3g/kg)、三七组(3g/kg)、田黄方组(3g/kg),采用高脂乳剂灌胃法复制高脂血症大鼠模型并给予相应药物治疗,给药30天后,采集24 h尿样用于代谢组学的分析。尿液样品UPLC-TOF/MS检测分析,质谱数据采MarkerLynx XS软件统计分析,并采用主成分分析法(PCA)和偏最小二乘-判别分析法(PLS-DA)对获得的数据进行处理。结果:在得分图中,各给药组与模型组显著分离;黄连组与三七组较接近,处于模型组和正常组之间;田黄方组和正常组较接近。同时三七、黄连、田黄方分别对8、10、15个生物标志物具有干预作用,综合评价各给药组对高脂血症大鼠代谢和生物标志物的干预作用,结果田黄方全方作用最好,黄连次之,三七最弱。结论:田黄方(3g/kg)对饮食性高脂血症大鼠模型的干预作用主要体现在对脂肪代谢、氨基酸代谢、炎症和氧化应激的调节作用,而三七、黄连虽能一定程度的改善模型组大鼠的代谢紊乱状态,但未能从根本上恢复,说明田黄方组合的合理性。

Objective：On the basis of previous research found that tianhuang compound had better effect on treating hyperlipidemia than sanqi and huanglian,to futher study the compatibility rule of tianhuang compound based on urine metabonomics. Methods： Thirty male SD rats were randomly divided into five groups, including control group, model group, huanglian group, sanqi group and tianhuang compound group. Rat model of hyperlipidemia was established by intragastric administration of high fat emulsion, and then the modeled rats were given correspond- ing medicationrespectively. After treatment for 30 days, urine samples were collected for the establishment of metabolism fingerprint by ultra - performance liquid chromatography / quadrupole time - of- flight mass spectrometry（ UPLC/QTOF MS）. The differences of metabolism fin- gerprint in the above groups were studied by principal componentanalysis （PCA） and partial least squares - discriminant analysis （ PLS-DA）, and then biomarkers were screened by variable importance in the projection（VIP） and compared in various groups. Result：In the score plot, huanglian group, sanqi group were separated with model group, the tianhuang compound group was close to control group and far away from the model group, the sanqi group was close to huanglian group but far away from the control group. The order of effect strength was tianhuang compound, huanglian, sanqi. Conclusion：Sanqi mainly regulated amino acid and energy metabolism, huanglian played a role in regulating in- flammation, oxidative stress and energy metabolism,while tianhuang compound had an overall effect on the above metabolic disorders, indica- ted that compatibility rule of tianhuang compound was reasonable.