基于UHPLC-Q-TOF-MS/MS探究山茱萸酒制前后与黄芪相关组分配伍对糖尿病肾病大鼠血浆代谢组学的影响

Effect of compatibility of Corni Fructus before and after wine-processing with Astragali Radix on plasma metabolomics in diabetic nephropathy rats based on UHPLC-Q-TOF-MS/MS

基于炮制和配伍探究山茱萸与黄芪相关组分对糖尿病肾病大鼠血浆代谢组学的影响。SD大鼠随机分为4组,用高脂饲料联合30 mg·kg^(-1)链脲佐菌素(STZ)诱导糖尿病肾病大鼠模型。采用HE、PAS和Masson染色对各组大鼠的肾脏组织切片进行病理学观察。超高效液相色谱-四极杆飞行时间串联质谱(UHPLC-Q-TOF-MS/MS)代谢组学方法研究山茱萸酒制前后与黄芪相关组分配伍给药后对糖尿病肾病大鼠血浆代谢物的影响。给予药物治疗后,糖尿病肾病大鼠的肾组织损伤情况和间质胶原纤维沉积面积均有不同程度的改善(P<0.001)。血浆代谢组学的检测结果显示,在患病大鼠中共鉴定出71种与糖尿病肾病发病机制相关的生物标志物,主要涉及亚油酸代谢,咖啡因代谢,甘油磷脂代谢,苯丙氨酸、酪氨酸和色氨酸生物合成,花生四烯酸代谢,苯丙氨酸代谢,视黄醇代谢,乙醚脂质代谢。经药物干预后,其中有26种发生显著性回调,以精准炮制药对(P-CG_(5))疗效更好。该研究从血浆代谢组学的角度阐述了P-CG_(5)可从苯丙氨酸、酪氨酸和色氨酸生物合成,苯丙氨酸代谢,咖啡因代谢等途径改善糖尿病肾病的代谢紊乱,为山茱萸与黄芪相关组分配伍在中医临床上的应用提供了理论支撑和实验依据。

Based on the processing and compatibility,this study explored the effects of components in Corni Fructus(CF)and Astragali Radix(AR)on plasma metabolomics in diabetic nephropathy rats.SD rats were randomly divided into four groups and diabetic nephropathy rat model was induced by high-fat diet combined with 30 mg·kg^(-1)streptozotocin(STZ).Histopathological observations of kidney tissue sections of rats in each group were conducted using HE,PAS,and Masson staining.Ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS/MS)metabolomics method was employed to investigate the effects of CF before and after wine-processing combined with AR-related components on plasma metabolites in diabetic nephropathy rats.After drug treatment,kidney tissue damage and interstitial collagen fiber deposition area in diabetic nephropathy rats were improved to varying degrees(P<0.001).The detection results of plasma metabolomics showed that 71 biomarkers related to the pathogenesis of diabetic nephropathy were identified in diseased rats,mainly involving linoleic acid metabolism,caffeine metabolism,glycerophospholipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,arachidonic acid metabolism,phenylalanine metabolism,retinol metabolism,and ether lipid metabolism.After drug intervention,26 of them were significantly downregulated,with better efficacy observed in precision processed herb-pair group(P-CG_(5)).This study elucidated from the perspective of plasma metabolomics that P-CG_(5)could improve metabolic disorders in diabetic nephropathy through pathways such as phenylalanine,tyrosine and tryptophan biosynthesis,phenylalanine metabolism,and caffeine metabolism,providing theoretical support and experimental basis for the clinical application of CF and AR compatibility in traditional Chinese medicine.