基于^1H-NMR的平乐正骨活血灵抗凝血作用代谢组学研究

Metabolomics research of anticoagulant effect of Pingle Zhenggu Huoxue Ling based on the ~1H-NMR

目的:采用~1H-NMR代谢组学技术研究平乐正骨活血灵(简称活血灵)干预创伤性深静脉血栓(DVT)大鼠后的代谢产物,探究其作用机制。方法:将90只大鼠随机分为空白对照组、模型组、阿司匹林组、活血灵组、NF-κB抑制剂(PDTC)组、PDTC+活血灵组,分别编号为A组、B组、C组、D组、E组、F组。A、B组予以蒸馏水灌胃,C组予以阿司匹林灌胃,D组予以活血灵灌胃,E组予以PDTC灌胃,F组为PDTC+活血灵灌胃。灌胃给药第7天,除A组,均予以DVT造模,造模后每日继续予以相应药物灌胃。第14天进行大鼠眼眶取血,采用~1H-NMR技术分析大鼠血液代谢产物。采用主成分分析和正交偏最小二乘法判别分析,研究D、E、F组间代谢物差异,筛选DVT治疗的可能代谢物。结果:与D组比较,F组组别代谢差异物为丙氨酸、β-羟丁酸、乙酰肉碱、葡萄糖。与E组比较,F组组别代谢差异产物为β-羟丁酸、乙酰肉碱、葡萄糖、丙氨酸、苯丙氨酸。结论:实验中发现的丙氨酸、β-羟丁酸、乙酰肉碱、葡萄糖、苯丙氨酸可能是活血灵作用于机体后的重要标志物。

Objective:To study the mechanism of the metabolic product of Pingle Zhenggu Huoxue Ling intervention in DVT rats by ~1H-NMR metabolomics.Methods:A total of 75 rats were randomly divided into blank control group,model group,Aspirin group,Huoxue Ling group,PDTC group,PDTC+Huoxue Ling group,respectively numbered as group A,group B,group C,group D,group E,group F.Group A and group B were given distilled water,group C was given Aspirin by gavage,group D was given the Huoxue Ling,group E was given PDTC,group F was given PDTC+Huoxue Ling.All rats were administrated by gavage.In the 7th day of gavage,DVT was used in addition to group A,and the corresponding medicine was continued daily after the modelwas successful.On the 14th day,blood was collected from the orbit,using ~1H-NMR technology analysis of rat blood metabolites,using principal component analysis and orthogonal partial least squares discriminant analysis,D,E,F group differences metabolites,screening of DVT treatment may metabolites,to explore the mechanism of the prevention and treatment of DVT.Results:Compared with group D,the differences metabolites between group D and group F were alanine,β-hydroxybutyrate,acetylcarnitine and glucose.Compared with group E,the differences metabolites between the group E and group F were β-hydroxybutyrate,acetylcarnitine,glucose,alanine and phenylalanine.Conclusion:The alanine,β-hydroxybutyrate,acetylcarnitine,glucose and phenylalanine found in the experiment may be important markers of huoxuling.