制草乌肝毒性效应的代谢通路分析

Analysis of Metabolic Pathway of Hepatotoxicity Effect of Prepared Aconiti

目的基于代谢组学技术检测制草乌对大鼠肝功能指标及肝组织病理影响从而探究代谢通路分析。方法以制草乌为示例药物,以大鼠为研究对象,将大鼠按体重随机分成4组。根据预试验结果设定正常组、制草乌高剂量组、中剂量组、低剂量组。除正常组外,其余各给药组灌胃相应的药物溶液,1次/d,连续灌胃28 d,末次给药后收集血清样本,检测各组的血清生化指标及采用LC-MS非靶向代谢组学技术对血清内源性代谢产物变化进行分析。结果血清生化指标结果显示,与正常组比较,制草乌高剂量组ALT含量显著升高,有统计学意义(P<0.05);制草乌中和低剂量组指标差异无统计学意义(P>0.05);肝组织病理检测显示,正常组肝小叶结构清晰,肝细胞索和肝血窦大致呈放射状排列,肝细胞圆润、饱满,肝板排列规则、整齐。制草乌高剂量组肝细胞广泛水肿、空泡化、丧失结构完整性;制草乌中和低剂量组未见明显异常;从血清中共筛选出34个内源性差异代谢物,其中上调15个、下调19个,多数涉及烟酸和多种氨基酸代谢。结论明确制草乌具有一定的肝毒性;肝毒性机制主要与烟酰胺(NA)和1-甲基烟酰胺(MNA)代谢有关。

Objective:To explore the metabolic pathway analysis based on the metabolomics technique to detect the effects of prepared Aconiti on liver function indexes and liver histopathology in rats.Methods:The rats were randomly divided into 4 groups according to their body weight.According to the pre-test results,normal group,high dose group,medium dose group and low dose group were set up.In addition to the normal group,the other drug administration groups were given corresponding drug solution once a day for continuous gavage for 28 days.After the last administration,serum samples were collected to detect serum biochemical indexes and the changes of serum endogenous metabolites were analyzed by LC-MS non-targeted metabolomics technology.Results:The results of serum biochemical indexes showed that ALT content in high dose group was significantly increased compared with that in the normal group,with statistical significance(P<0.05);There was no significant difference between the two groups(P>0.05).Liver histopathological examination showed that in the normal group,the structure of hepatic lobules was clear,hepatocyte cords and hepatic blood sinuses were roughly arranged radially,hepatocytes were round and full,and hepatic plates were arranged regularly and neatly.Hepatocyte edema,vacuolation and loss of structural integrity were widespread in the high dose group.No obvious abnormality was found in the middle and low dose groups.A total of 34 endogenous differential metabolites were screened from serum,including 15 up-regulated and 19 down-regulated,most of which involved niacin and various amino acid metabolism.Conclusion:It is clear that prepared Aconiti has certain hepatotoxicity and the mechanism of hepatotoxicity is mainly related to the metabolism of niacinamide(NA)and 1-methylnicotinamide(MNA).