基于^(1)H-NMR代谢组学的柴胡醋制前后的毒性作用比较研究

Toxicity comparison of raw and vinegar-processed Bupleuri Radix based on ^(1)H-NMR metabolomics

该研究基于核磁共振氢谱(1H-NMR)比较柴胡醋制前后及不同醋炮制对柴胡毒性的影响,初步探讨其毒性作用机制.实验将32只SD大鼠随机分为4组:空白组(蒸馏水)(control)、柴胡生品组(15 g·kg^(-1)·d^(-1))(RB)、米醋制柴胡组(15 g·kg^(-1)·d^(-1))(R-VBRB)、陈醋制柴胡组(15 g·kg^(-1)·d^(-1))(S-VBRB),给药30 d,处理后观察病理切片,检测与肝、肾功能相关的生化指标,取大鼠血清、肝脏、肾脏进行1H-NMR数据检测,利用主成分分析法(principal component analysis,PCA)和正交偏最小二乘法判别分析(orthogonal partial least squares-discrimination analysis,OPLS-DA)进行分析处理.结果显示,与空白组相比,柴胡生品组ALT、AST和ALP显著升高,而醋制柴胡组ALT和ALP显著降低(P<0.05),说明柴胡具有一定的肝毒性,且醋制后能降低柴胡的肝毒性.肾功能相关的生化指标BUN和CREA在空白组和不同给药组中无显著差异,说明柴胡及醋制品对肾功能影响较小.多元统计分析结果显示,柴胡生品影响肝脏代谢的标志物包括甲硫氨酸、谷氨酰胺和谷氨酸等,影响肾脏代谢的标志物包括牛磺酸、鸟氨酸、肌醇等,主要参与氨基酸代谢、能量代谢、脂质代谢和牛磺酸代谢等.柴胡醋制后对这些标志物的影响程度均有降低趋势,且陈醋制柴胡的影响较小.综合其生化指标及代谢组学分析,柴胡生品及醋制品对大鼠均有一定的毒性作用,且醋制能降低其毒性,其中陈醋制柴胡对肝肾代谢的影响小于米醋制柴胡,说明炮制用醋对柴胡的毒性也有一定的影响.

This study compared the toxicity of raw Bupleuri Radix(BR)and vinegar-processed Bupleuri Radix(VPBR)based on proton nuclear magnetic resonance(~1H-NMR),and explored the mechanism of toxicity.Thirty-two male Sprague-Dawley(SD)rats were randomly divided into four groups:a control group(distilled water),a raw BR group(15 g·kg^(-1)·d^(-1)),a rice VPBR(R-VPBR)group(15 g·kg^(-1)·d^(-1)),and a shanxi VPBR(S-VPBR)group(15 g·kg^(-1)·d^(-1)).After administration for 30 d,pathological sections were treated and observed,and biochemical indexes related to liver and renal function were determined.The serum,liver,and kidney of rats were collected and analyzed by~1H-NMR.The principal component analysis(PCA)and orthogonal partial least squares-discrimination analysis(OPLS-DA)were performed.The results showed that,as compared with the control group,alanine aminotransferase(ALT),aspartate aminotransferase(AST),and alkaline phosphatase(ALP)in the raw BR group were increased significantly,while ALT and ALP in the R-VPBR and S-VPBR groups were significantly decreased(P<0.05),which indicated that BR showed certain hepatotoxicity,and vinegar processing reduced its hepatotoxicity.No significant difference of blood urea nitrogen(BUN)and creatinine(CREA),the biochemical indexes related to renal function,was observed in the control group and administration groups,indicating that BR had less effect on the renal function.The results of multivariate statistical analysis showed that the biomarkers of BR affecting liver metabolism were methionine,glutamine,and glutamic acid,and affecting kidney metabolism were taurine,ornithine,and inosine.These biomarkers were mainly involved in amino acid metabolism,energy metabolism,lipid metabolism,and taurine metabolism.VPBR alleviated the effect on the biomarkers,and S-VPBR had smaller effect than R-VPBR.Combining the results of biochemical indexes and metabolomics analysis,both raw BR and VPBR showed toxic effect on rats,whereas vinegar processing reduced its toxicity.S-VPBR has smaller effect on kidney and liver metabolism than R-VPBR,which indicates that the vinegar used for processing has certain effect on the toxicity of BR.