摘要目的 基于气相色谱-质谱联用技术(GC-MS)测定脓毒症大鼠血浆代谢组学指标,寻找脓毒症差异代谢产物,初步揭示脓毒症的病理生理变化.方法 将8周龄雄性SD大鼠按随机数字表法分为假手术组(n=18)和脓毒症组(n=24).采用盲肠结扎穿孔术(CLP)制备脓毒症大鼠模型;假手术组只游离盲肠,不予以结扎和穿孔.制模后6 h取心脏血,通过GC-MS测定两组血浆中的小分子代谢产物,并对其进行代谢轮廓分析,筛选相关差异代谢产物.结果 主成分分析(PCA)和偏最小二乘-判别分析(PLS-DA)显示,脓毒症组与假手术组大鼠血浆代谢谱差异显著.GC-MS共筛选出259个代谢物,脓毒症组与假手术组之间有69个差异代谢产物,其中23个与氨基酸代谢有关;脓毒症组血浆中腐胺、N-氨甲酰谷氨酸、羟基正缬氨酸、3-氰丙氨酸、D-丙氨酰-D-丙氨酸、尿素含量较假手术组明显升高,脓毒症组/假手术组含量比值分别为10.876、6.394、2.800、2.226、1.323、1.203(均P<0.05);而脓毒症组血浆中氧代脯氨酸、瓜氨酸、谷氨酰胺、苏-β-羟基天门冬氨酸、柠檬酸、N-甲基-DL-丙氨酸、丝氨酸、赖氨酸、苏氨酸、N-甲酰-L-甲硫氨酸、甲硫氨酸、丙氨酸、烟酰甘氨酸、N-甲基-L-谷氨酸、反式-4-羟基-L-脯氨酸、脯氨酸、L-谷氨酸含量较假手术组明显降低,脓毒症组/假手术组含量比值分别为0.858、0.853、0.834、0.816、0.816、0.814、0.813、0.801、0.793、0.792、0.774、0.766、0.748、0.727、0.716、0.674、0.603(均P<0.05).结论 用GC-MS检测脓毒症大鼠代谢组学指标显示能量代谢异常;血浆氨基酸含量可以评估脓毒症时的能量代谢状态.

abstractsObjective To reveal the pathophysiological changes of sepsis, the plasma metabonomics of septic rats was determined and differential metabolites were determined by gas chromatography-mass spectrometry (GC-MS). Methods Male Spraguee-Dawley (SD) rats about 8 weeks were randomly divided into sham group (n = 18) and sepsis group (n = 24). Cecal ligation and puncture (CLP) was used to build sepsis model, while cecum was kept intact only in the sham group. 6 hours after the operation, rats were anesthetized, and blood was harvested through heart thoracotomy. Then the plasma metabonomics was detected by GC-MS and metabolic profile analysis was performed to find the relative differential metabolites.Results Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that the metabolic profiling of the sepsis group was significantly different from the sham group. 259 kinds of metabolites were got by GC-MS, and 69 kinds of differential metabolites were found between sham group and sepsis group, in which 23 differential metabolites were related to amino acid metabolism. Compared with sham group, the contents of putrescine, N-glutamic acid, hydroxynorvaline, 3-cyanuric acid, D-alanyl-D-alanine and urea in the plasma of septic rats increased significantly, which ratios of sepsis/sham group were 10.876, 6.394, 2.800, 2.226, 1.323, 1.203, respectively (allP < 0.05). On the other hand, the contents of oxygen generation of proline, citrulline, glutamine, su-beta-hydroxy aspartic acid, citric acid, N-methyl-DL-alanine, serine, lysine, threonine, N-formyl-L-methionine, methionine, alanine, nicotinuric acid, N-methyl-L-glutamic acid, trans-4-hydroxy-L-proline, proline, L-glutamic acid in the plasma of septic rats decreased significantly, which ratios of sepsis/sham group were 0.858, 0.853, 0.834, 0.816, 0.816, 0.814, 0.813, 0.801, 0.793, 0.792, 0.774, 0.766, 0.748, 0.727, 0.716, 0.674, 0.603, respectively (allP < 0.05).Conclusions Through the GC-MS analysis of plasma metabonomics of septic rats, we found abnormal energy metabolism changes. The content of amino acid in plasma might be a method to evaluate the energy metabolism status of sepsis.