何首乌致肾阳虚模型大鼠肝损伤的血浆代谢组学研究

Plasma metabolomics study on liver injury in kidneyyang deficiency syndrome rats induced by raw and processed Polygonum multiflorum

目的基于代谢组学研究策略,研究何首乌给药后肾阳虚大鼠血浆中内源性代谢物的变化,寻找具显著性差异的生物标志物,探讨何首乌致大鼠肝损伤的作用机理。方法大鼠皮下注射氢化可的松建立肾阳虚模型,分别给予大鼠高(24 g·kg^(-1))、中(12 g·kg^(-1))、低(6 g·kg^(-1))剂量的制首乌和生首乌(12 g·kg^(-1))。以超高效液相色谱质谱联用(UPLC-MS)技术为核心技术,以OPLS方法为数据分析手段,研究大鼠血浆内源性代谢谱的变化。结果找到8个与肝损伤相关的潜在生物标志物(含一个未知化合物)。给予肾阳虚大鼠生首乌后,与肾阳虚模型组比较,大鼠体内的肉毒碱、LPC0∶0/16∶0、LPC16∶0/0∶0、LPC0∶0/18∶0和未知物22.5_269.0水平显著下调,缬氨酸、苯丙氨酸和色氨酸含量显著增加;制首乌高、中剂量组中,大鼠体内的以上8种代谢物亦有相同变化趋势,但显著性程度略有不同;低剂量组中相关代谢物的强度虽然发生变化,但与模型组相比多无统计学意义,仅LPC0∶0/16∶0和LPC0∶0/18∶0有显著性变化。结论生首乌对肾阳虚大鼠有肝毒性,制首乌对肾阳虚大鼠有一定的肝损伤,何首乌对肝脏功能的影响与给药剂量和炮制方法有一定相关性。何首乌致大鼠肝损伤的机制可能与氨基酸代谢和免疫抑制有关。

Objective To study the changes of endogenous metabolites in plasma of kidney-yang deficiency syndrome rats after administration of Polygonum multiflorum,identify the potential biomarkers and explore the mechanism of liver injury induced by Polygonum multiflorum based on the metabolomics strategies.Methods The Kidney-yang deficiency syndrome model was established by subcutaneous administration of hydrocortisone,after which the rats were administrated with three dose of processed Polygonum multiflorum（ PP） and rawPolygonum multiflorum（ RP）. And then,a UPLC-MS based metabolomic method was developed for investigating the metabolic disturbances caused by RP and PP,followed by the pattern recognition approach. Results Eight potential biomarkers in correlation with liver injury were identified including one unknown compound. Comparing with the model group,the level of carnitine,LPC0： 0/16： 0,LPC16： 0/0： 0,LPC0： 0/18： 0 and unknown compound（ tR m/z 22. 5_269. 0） were decreased,while those of valine,phenylalanine and tryptophan in RP group were increased considerably. The level of those biomarkers in PP-H group and PP-Mgroup had the same trend,but the degree of significance was slightly different.Although the intensity of the metabolites in the PP-L group changed,it was not statistically significant compared with the model group except LPC0： 0/16： 0 and LPC0： 0/18： 0. Conclusion RP has a liver toxicity in rats with kidney-yang deficiency syndrome,PP has some liver injury on model rats. The effect of Polygonum multiflorum on liver is related to the dose and processing method. The mechanism of the liver injury caused by Polygonum multiflorum may be related to amino acid metabolism and immunosuppression.