代谢组学测定对急性百草枯中毒大鼠的判定作用  

Diagnostic effect of metabonomics determination method on acute paraquat poisoning

作　　者：宋偲婷 刘智玲[1] 张义雄[1] 张先瑞[2] 陈柳乔 

机构地区：[1]湖南师范大学第一附属医院,湖南省人民医院急诊科,湖南长沙410005 [2]湖南师范大学第一附属医院,湖南省人民医院药剂科,湖南长沙410005 [3]湖南师范大学第一附属医院,湖南省人民医院科教科,湖南长沙410005

出　　处：《中华危重病急救医学》2016年第4期329-333,共5页Chinese Critical Care Medicine

基　　金：湖南省卫生和计划生育委员会科研计划项目（B2016009）

摘　　要：目的采用气相色谱-质谱联用（GC—MS）代谢组学技术筛选急性百草枯（PQ）中毒大鼠血浆中的潜在生物标志物，为早期诊断提供依据。方法8只SD大鼠按随机数字表法分为PQ中毒组（灌胃PQ溶液100mg／kg）及对照组（灌胃等量生理盐水），每组4只。于灌胃后2、24、48h观察大鼠一般情况；收集大鼠眼眶血，采用GC—MS方法检测血浆中内源性小分子代谢物，并进行代谢轮廓及随机森林分析，筛选潜在的生物标志物。结果①PQ中毒组大鼠染毒后逐渐出现少动、呼吸急促、腹部抽动等中毒症状；对照组大鼠则生命体征平稳。②利用GC—MS技术分析大鼠血浆中代谢物后，结合主成分分析（PCA）及偏最小二乘判别式分析（PLS—DA）模型图，得出PQ中毒组各时间点大鼠血浆代谢分布较分散，而对照组分布比较密集，说明两组大鼠体内产生的代谢模式不同；而且PQ中毒组大鼠灌胃2h起代谢轨迹即较对照组明显偏移，至48h其轨迹与对照组接近，表明大鼠在中毒早期血浆代谢产物已发生了明显差异。采用随机森林方法筛选出5种权重较大的潜在生物标志物，分别为丝氨酸、L-天冬酰胺、棕榈酸、硬脂酸、花生四烯酸，其保留时间分别为15．259、24．345、33．334、37．695、40．254min。PQ中毒组丝氨酸、L-天冬酰胺、花生四烯酸含量较对照组显著升高，分别于48h、48h、24h达峰值（40．88±5．38比28．85±2．32、6．61±1．31比0．76±0．65、14．21±4．28比4．42±1．19，均P〈0．01）；棕榈酸和硬脂酸的含量则较对照组显著降低，均于48h达谷值（39．09±10．23比83．99±20．49、44．03±3．60比140．76±73．91，P〈0．05和P〈0．01），提示这些代谢物的改变与PQ毒性有关，说明PQ会干扰大鼠体内的能量代谢及脂质代谢。结论代谢组学分析筛选出PQ中毒大鼠血浆丝氨酸、L-天冬酰胺、花生四烯酸含量�Objective To screen the potential biomarkers in plasma of rats with acute paraquat （PQ） poisoning using gas chromatography-mass spectrometry （GC-MS） based metabonomics technology, and to provide concrete evidence for early diagnosis. Methods Eight Sprague-Dawley （SD） rats were randomly divided into PQ poisoning group （intragastricly administrated with PQ solution 100 mg/kg） and control group （intragastricly administrated with the same volume of normal saline） according to the random number table, with 4 rats in each group. The general situation of rats was observed at 2, 24 and 48 hours after administration. The blood of eye sockets was collected, the endogenous small molecule metabolites in plasma were determined with GC-MS method, and metabolic profile analysis and random forest analysis were performed to filter the potential biomarkers. Results ① The rats in PQ poisoning group gradually appeared lack movement, tachypnea, abdominal seizure and other symptoms of poisoning. In control group, the vital signs were stable. ② The metabolites in plasma of rat were analyzed with GC-MS analysis, and the diagrammatic figure was plot as combined with principal component analysis （PCA） and partial least squares-discriminated analysis （PLS-DA） model, which showed that the distribution of plasma metabolism in PQ poisoning group was more diffuse but in the control group was more intensive, indicating that the metabolic patterns in two groups were different, From 2 hours after PQ administration, the metabolic trajectory in PQ poisoning group was significantly deflected compared with that of the control group, which was similar to control group until 48 hours, indicating that the metabolites in plasma of rat showed obvious difference in the early period. Five kinds of potential biomarkers with large weights were selected by random forest method which were serine, L-asparagine, hexadecanoic acid, octadecanoic acid, and arachidonic acid, the retention time was 15.259, 24.345, 33.334, 37.695, and 40

关 键 词：百草枯 中毒 代谢组学 气相色谱-质谱分析 

分 类 号：R595.4[医药卫生—内科学]