温肾降浊散治疗大鼠C-BSA肾炎模型的代谢组学研究

Metabonomics study of Wenshen Jiangzhuo powder in the treatment of C- BSA nephritis model in rats

目的:通过代谢组学方法探究温肾降浊散治疗大鼠阳离子化牛血清白蛋白(C-BSA)肾炎模型的作用机理。方法:以GC-MS法测定温肾降浊散治疗前后C-BSA肾炎模型大鼠血清中内源性代谢产物,计算其含量变化,结合主成分分析法对这一变化过程进行模式识别。结果:确定了血清中39种代谢产物,经方法学考察,其精密度良好,制备后的样品在12 h内稳定,方法重复性良好。对内源性代谢物数据进行PCA主成分分析、OPLS-DA分析和t检验分析,确定了15个具有显著贡献率的潜在生物标记物;与正常组比较,模型组大鼠血清中乳酸、苹果酸、棕榈酸、亚油酸、反油酸、花生四烯酸、1-磷酸肌醇、胆固醇、2-单棕榈酸甘油酯、1-单硬脂酸甘油酯的相对峰面积均明显升高,L-亮氨酸、L-缬氨酸、氨基丙二酸则有所降低,硬脂酸与花生酸则无明显变化;与模型组比较,温肾降浊散组大鼠血清中乳酸、棕榈酸、亚油酸、反油酸、花生四烯酸、1-磷酸肌醇、胆固醇、2-单棕榈酸甘油酯、1-单硬脂酸甘油酯的相对峰面积均有回落,L-亮氨酸、L-缬氨酸、氨基丙二酸则有所回升,而苹果酸、硬脂酸与花生酸则无明显变化。结论:温肾降浊散可通过改善肾小球通透性,提高自由氨基酸浓度及肾脏清除率等方式来调节机体脂代谢、氨基酸代谢和糖代谢紊乱,进而使肾脏恢复正常功能。

Objective： To study the mechanism of Wenshen Jiangzhuo powder in the treatment of cationic bovine serum albumin（ C- BSA） nephritis model in rats by metabonomic method. Methods： The endogenous metabolites in serum of rat C- BSA nephritis model were determined by GC- MS method before and after the treatment of Wenshen Jiangzhuo powder,the changes in the content of metabolites were calculated,and the pattern of the process of the change was recognized in combination with the principal component analysis method. Results： Totally 39 metabolites in serum were determined. The methodology validation showed the precision was good and the prepared sample was stable within 12 h with good repeatability. The endogenous metabolite data were analyzed by principal component analysis（ PCA）,OPLS- DA analysis and t- test,and 15 potential biomarkers of significant contribution rate were identified. Compared with the normal group,the relative peak areas of lactic acid,2- hydroxybutanedioic acid,hexadecanoic acid,9,12- octadecadien- oic acid,11- trans- octadecenoic acid,arachidonic acid,1- phosphate inositol,cholesterol,2- monopalmitin ether and 1- monostearin ether in serum of rats in the model group significantly increased,the relative peak areas of L- leucine,L- valine and aminomalonic acid decreased,and the relative peak areas of octadecanoic acid and eicosanoic acid had no obvious change. Compared with the model group,the relative peak areas of lactic acid,hexadecanoic acid,9,12- octadecadien- oic acid,11- trans- octadecenoic acid,arachidonic acid,1- phosphoinositide,cholesterol,2- monopalmitin ether and 1- monostearin ether in serum of rats receiving Wenshen Jiangzhuo powder all decreased,the relative peak areas of L- leucine,L- Valine and aminomalonic acid rebounded,and the relative peak areas of 2- hydroxybutanedioic acid,octadecanoic acid and eicosanoic acid showed no significant change. Conclusion： Wenshen Jiangzhuo powder can regulate the body＇s fat metabolism,amino acid metabolism and glucose metabolism disorders by improving the glomerular permeability,increasing the free amino acid concentration and renal clearance,and thus restoring the normal function to the kidney.