基于代谢组学技术筛选大鼠低温失血性休克的预警指标

Research on screening early warning indicators of hypothermia hemorrhagic shock in the rats based on metabolomics technology

目的 基于代谢组学技术筛选大鼠低温失血性休克预警指标。方法 30只SD大鼠随机分成对照组、低温失血25%组和低温失血50%组,每组10只。对照组大鼠于正常环境中(22±2)℃饲养,低温失血25%组和低温失血50%组大鼠于低温环境(-10±2)℃中饲养。采用定容性失血性休克的模型建立方法,监测失血总血量25%和50%时生命体征和凝血功能变化;收集失血总血量25%和50%后血浆,使用LC-MS/MS筛选代谢物差异表达。结果 生命体征监测发现,相较于对照组,低温失血25%组和低温失血50%组体温分别下降到(37.60±0.12)℃和(35.10±0.19)℃,心率分别升至(389.0±5.3)次/min和(438.0±7.2)次/min,平均动脉压降至(69.0±6.1)mmHg和(40.0±5.1)mmHg,氧分压(PaO_(2))降至(100.5±2.1)mmHg和(79.5±1.8)mmHg,二氧化碳分压(PaCO_(2))升至(51.1±2.0)mmHg和(72.3±1.7)mmHg,差异均有统计学意义(P<0.05)。凝血功能检测发现,与对照组相比,失血25%和失血50%时活化部分凝血活酶时间(APTT)升至(25.3±1.1)s和(32.3±1.7)s,血浆凝血酶原时间(PT)升至(16.4±0.9)s和(23.1±2.1)s,组织型纤溶酶原激活剂(t-PA)升至(4.7±0.6)g/L和(5.9±1.0)g/L,差异均有统计学意义(P<0.05);纤溶酶原激活剂抑制物-1(PAI-1)维持在(1.6±0.4)μg/L和(1.6±0.7)μg/L,差异无统计学意义(P>0.05);代谢组学发现,大鼠低温失血后血浆中创伤激素、牛去氧胆酸盐、4-氨基丁酸等代谢物具有明显变化。结论 大鼠随失血量增加,生命体征和凝血功能发生进行性改变,基于代谢组学筛选的差异代谢物,可为低温失血性休克预警提供参考。

Objective To use metabolomics technology to screen early warning indicators of hypothermia hemorrhagic shock in the rats.Methods Thirty SD rats were randomly divided into a control group,a 25%hypothermia hemorrhagic shock group and a 50%hypothermia hemorrhagic shock group,with 10 rats in each group.The rats in the control group were raised in a normal environment(22±2)℃,while the rats in the 25%and 50%hypothermia hemorrhagic shock groups were raised in a low temperature environment(-10±2)℃.A model of controlled hemorrhagic shock was established,and the changes in vital signs and coagulation function were monitored when the total blood loss reached 25%and 50%.Plasma samples were collected after 25%and 50%blood loss,and the metabolites with differential expression were screened by using LC-MS/MS.Results Vital sign monitoring revealed that compared with the control group,the body temperature in 25%and 50%hypothermia hemorrhagic shock groups decreased to(37.60±0.12)℃and(35.10±0.19)℃respectively,heart rate increased to(389.0±5.3)beats/min and(438.0±7.2)beats/min,mean arterial pressure decreased to(69.0±6.1)mmHg and(40.0±5.1)mmHg,arterial partial pressure of oxygen(PaO_(2))decreased to(100.5±2.1)mmHg and(79.5±1.8)mmHg,and arterial partial pressure of carbon dioxide(PaCO_(2))increased to(51.1±2.0)mmHg and(72.3±1.7)mmHg(P<0.05).Coagulation function testing revealed that compared with the control group,activated partial thromboplastin time(APTT)in 25%and 50%hypothermia hemorrhagic shock groups increased to(25.3±1.1)s and(32.3±1.7)s,the prothrombin time(PT)increased to(16.4±0.9)s and(23.1±2.1)s,the tissue-type plasminogen activator(t-PA)increased to(4.7±0.6)g/L and(5.9±1.0)g/L(P<0.05),while the plasminogen activator inhibitor-1(PAI-1)remained at(1.6±0.4)μg/L and(1.6±0.7)μg/L,there was no statistical difference(P>0.05).Metabolomics analysis revealed significant changes in metabolites such as traumatic-acid,taurodeoxycholate,and 4-aminobutanoate in the plasma.Conclusions As the amount of blood loss increases,there are progressive changes in vital signs and coagulation function in rats.Differential metabolites identified by metabolomics screening can provide the references for early warning of hypothermia hemorrhagic shock.