基于代谢组学技术的毒鼠强中毒机制研究

Reserch on the mechanism of Tetramine poisoning based on metabolomics technology

目的研究毒鼠强中毒后大鼠的血浆代谢组学随时间变化规律,从代谢组学角度揭示其毒理学作用机制,为毒鼠强中毒的相关法医学研究提供依据。方法将SD大鼠随机分成对照组和实验组(n=8),实验组大鼠灌胃1/4LD50(0.05 mg/kg)毒鼠强乙酸乙酯溶液,对照组灌胃等剂量乙酸乙酯溶液,灌胃后于24 h、5 d、16 d眼眶静脉采血(约1 mL),应用超高效液相色谱-飞行时间质谱联用技术检测血浆中的内源性小分子物质。采用主成分分析(PCA)、正交偏最小二乘判别分析(OPLS-DA)等筛选显著差异代谢物,寻找与毒鼠强中毒相关的代谢通路。结果本研究以变量权重重要性排序(VIP)值>1和student t检验的P值<0.05为筛选标准,中毒后不同时间点共筛选出显著差异代谢物30种,包括鞘脂类、氨基酸类和脂肪酸类化合物,涉及到鞘脂信号通路、鞘脂代谢和ABC蛋白转运等10条代谢通路。结论毒鼠强中毒后机体多条代谢通路受到显著影响,主要是不同程度的神经系统调节紊乱和能量代谢紊乱,整体变化呈现出累积效应,可为研究毒鼠强毒性作用机制提供依据。

Objective To study the changes of plasma metabolomics in rats with tetramine poisoning over time,and to reveal its toxicological mechanism from the perspective of metabolomics,so as to provide evidence for related forensic research.Methods SD rats were randomly divided into control group(n=8)and experimental group(n=8).Rats in experimental group were given 1/4LD50(0.05 mg/kg)tetramine solution,while rats in control group were given equal dose of ethyl acetate solution.Blood samples(about 1 mL)were collected from orbital veins on 24 h,5 d,16 d,and the significantly small molecules in plasma were investigated by ultra-performance liquid chromatography-time-of-flight mass spectrometry.The principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLSDA)were used to process data,to screen significantly different metabolites,and to search for metabolic pathways related to tetramine poisoning.Results According to the variable importance in the projection(VIP)>1 and student’s t-test P<0.05,we found thirty different metabolites in this experiment,including sphingolipids,amino acids and fatty acids compounds.10 metabolic pathways were involved,such as sphingolipid signaling pathway,sphingolipid metabolism and ABC transporter.Conclusion Multiple metabolic pathways are significantly affected after tetramine poisoning,mainly nervous system regulation disorder and energy metabolism disorder.The overall changes showed a cumulative effect,which can provide a certain basis for studying the mechanism of tetramine toxicity.