基于^1H NMR代谢组学细辛肝肾毒性的机制研究

Study on the Hepatotoxicity and Nephrotoxicity of Asarum in Rats Based on^1H-NMR Metabonomics

目的:运用代谢组学方法研究细辛对大鼠肝肾组织代谢学特征,阐明细辛肝肾毒性机制。方法:20只大鼠随机分为4组,每组5只,分别为空白对照组、细辛0.27 g/kg、0.81 g/kg、1.35 g/kg三个剂量组,给药28 d后,取肝肾组织样本,进行1H-NMR代谢组学分析,采用主成分分析及正交偏最小二乘判别分析法研究细辛各给药组与空白对照组之间的代谢谱差异,筛选潜在生物标志物,探讨细辛的肝肾毒性机制。结果:与空白对照组比较,细辛0.27 g/kg组肝组织代谢物无明显差异,细辛0.81 g/kg组有亮氨酸、醋酸、天冬氨酸、胆碱磷酸、甜菜碱、牛磺酸、β-葡萄糖7种化合物浓度显著增加(P<0.05),细辛1.35 g/kg组有亮氨酸、丙氨酸、醋酸、蛋氨酸、谷氨酸、天冬氨酸、肌酸酐、胆碱磷酸、甜菜碱、牛磺酸、α-葡萄糖、β-葡萄糖12种化合物浓度显著增加(P<0.05),丝氨酸、糖原2种化合物浓度显著下降(P<0.05);细辛各给药组肾组织中代谢物均有明显差异,其中细辛1.35 g/kg组异亮氨酸、亮氨酸、赖氨酸、丙氨酸、醋酸、脯氨酸、蛋氨酸、谷氨酸、琥珀酸、三乙胺、肌酸酐、胆碱、胆碱磷酸、甜菜碱、牛磺酸、肌醇、甘氨酸、酪氨酸、苯丙氨酸、α-葡萄糖、β-葡萄糖21种化合物均显著升高(P<0.05)。结论:细辛的肝肾毒性可能促进氨基酸代谢、能量代谢、脂质代谢等多种途径,增加机体氧化损伤、放大炎症效应而发生细胞毒性,与传统医学认为细辛辛温偏热促进机体代谢的观点一致。

Objective:To study the metabonomic characteristics of Asarum on liver and kidney tissues of rats and its hepatotoxicity and nephrotoxicity mechanism.Methods:20 rats were randomly divided into four groups,5 in each group:the control group and three doses of Asarum groups(0.27 g/kg,0.81 g/kg,1.35 g/kg).After 28 days of administration,samples of liver and kidney tissues were collected for 1 H-NMR metabolism analysis.Potential biomarkers were screened by principle component analysis and Orthogonal partial least squares discriminant analysis on metabolism profiling of different groups to study the liver and kidney toxicity mechanism of Asarum.Results:Compared with the control group,there was no significant difference in liver metabolites in 0.27 g/kg drug group.In 0.81 g/kg Asarum group,7 kinds of potential biomarkers were increased(P<0.05),including leucine,acetate,aspartate,phosphocholine,betaine,taurine andβ-glucose.In 1.35 g/kg Asarum group,12 kinds of potential biomarkers were increased(P<0.05),including leucine,alanine,acetate,aspartate,methionine,glutamate,creatinine,phosphocholine,betaine,taurine,α-glucose andβ-glucose.2 kinds of potential biomarkers were decreased(P<0.05),including serine and glycogen.The metabolites in kidney tissues in all Asarum group had significantly difference.In 1.35 g/kg Asarum group,21 kinds of potential biomarkers were increased(P<0.05),including isoleucine,leucine,lysine,alanine,acetate,proline,methionine,glutamate,succinate,trimethylamine,creatinine,choline,phosphocholine,betaine,taurine,institol,glycine,tyrosine,phenylalanine,α-glucose andβ-glucose.Conclusion:The hepatotoxicity and nephrotoxicity of Asarum may promote the metabolism of glucose,amino acid,and lipid,increase the oxidative damage of the body,and amplify the inflammatory effects,which is consistent with the Xinwenpianre theory in traditional Chinese medicine.