大鼠脑干早期创伤性轴索损伤的非靶向代谢组学研究

A non-targeted metabolomic study of early traumatic axonal injury in rat brainstem

目的 通过非靶向代谢组学技术分析脑干创伤性轴索损伤后30 min的差异代谢物和关键代谢通路。方法将10只(8周龄,雄性)SD大鼠随机分为假手术组(Sham组)和脑干创伤性轴索损伤组(TAI组),每组5只。采用Marmarou模型建立SD大鼠脑干创伤性轴索损伤模型,并以生命体征测量、HE染色和嗜银染色进行模型评价。采用UPLC/TripleTOF-MS系统分析TAI组和Sham组脑干组织的代谢物变化,筛选差异代谢物和分析差异代谢通路。结果 与Sham组相比,TAI组被打击后立即进入不同程度的昏迷状态,伴有生命体征紊乱,伤后30 min发现脑干存在弥漫性轴索损伤。代谢组学分析筛选出差异代谢物包括月桂酰二乙醇酰胺、邻苯二甲酸单乙基己基酯、PE(20:5(5Z,8Z,11Z,14Z,17Z)/24:1(15Z))、PC(16:0/16:1(9Z))、(2E,6Z)-十二二烯酸甲酯、六甲密胺,涉及代谢途径包括甘油磷脂代谢、甘油脂代谢、花生四烯酸代谢、逆行内源性大麻素信号、自噬-动物、糖基磷脂酰肌醇(GPI)-锚定生物合成。结论 利用UPLC/TripleTOF-MS技术进行大鼠脑干早期TAI的代谢组学研究,发现了6个潜在生物标志物及相关代谢通路,为脑干早期TAI的法医学鉴定和诊治提供了依据。

Objective To analyse differential metabolites and key metabolic pathways 30 minutes after traumatic axonal injury of the brainstem by non-targeted metabolomics techniques. Methods Ten(8-week-old, male) SD rats were randomly divided into a sham-operated group(Sham group) and a traumatic axonal injury to the brainstem group(TAI group), with five rats in each group. The Marmarou model was used to establish the SD rat brainstem traumatic axonal injury model, and the model was evaluated by vital sign measurement, HE staining and silver plating staining. The UPLC/TripleTOF-MS system was used to analyse metabolite changes in brainstem tissues of the TAI and Sham groups, to screen for differential metabolites and to analyse differential metabolic pathways. Results Compared with the Sham group, the TAI group entered a coma of varying degrees immediately after the blow with disturbances in vital signs, and diffuse axonal damage was found in the brainstem 30 minutes after injury. Metabolomic analysis screened for differential metabolites including lauryl diethanolamide, monoethylhexyl phthalate, PE(20:5(5Z,8Z,11Z,14Z,17Z)/24:1(15Z)), PC(16:0/16:1(9Z)),(2E,6Z)-dodecadienoic acid methyl ester, hexamethylmethanamine, and involved metabolic pathways including retrograde endocannabinoid signaling, autophagy-animal, arachidonic acid metabolism, glycosylphosphatidylinositol(GPI)-anchored biosynthesis, glycerophospholipid metabolism and glycerolipid metabolism. Conclusion A metabolomic study of brain tissue from early TAI in rat brainstem using UPLC/TripleTOF-MS technique initially screened six potential biomarkers and identified relevant metabolic pathways, which provides a basis for the diagnosis, treatment and forensic identification of early TAI in brainstem.