摘要:
通过气质联用仪和液质联用仪的非靶向检测，结合主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)等方法，对日本沼虾肝胰腺进行代谢物分析，以研究在非致死剂量的亚硝酸盐或氨胁迫下，日本沼虾肝胰腺新陈代谢的变化。经亚硝酸盐(0.5 mg·L-1，以N计)胁迫2 d后，鉴定到具有显著差异的代谢化合物46个，涉及氨基酸代谢、脂肪酸代谢和甘油磷脂代谢等；pH=9.0时，氨(1.0 mg·L-1，以N计，即非离子氨氮为0.378 mg·L-1)胁迫后，鉴定出显著差异性代谢化合物52个，涉及三羧酸循环、氨基酸代谢和甘油磷脂等代谢通路。选择pH=9.0氨(1.0 mg·L-1)胁迫组对日本沼虾肝胰腺中的三羧酸循环进行定量验证实验，结果显示，日本沼虾肝胰腺中草酰乙酸含量与对照组相比在12 h显著升高，24 h和48 h后则与对照组无显著差异。α-酮戊二酸含量与对照组相比在12 h即有显著增加，24 h含量与对照组相比显著下降，48 h后与对照组相比无明显差异。推测pH=9.0时氨氮(1.0 mg·L-1)胁迫12 h即会对三羧酸循环产生影响。
Abstract:
In order to study the changes in the metabolism of hepatopancreas of Macrobrachium nipponensis under non-lethal doses of nitrite or ammonia stress, metabolite analysis was carried out by non-targeted detection of GC/MS and LC/MS, combined with the methods of principal component analysis (PCA) and partial orthogonal least squares discriminant analysis (OPLS-DA). After 2 days of stress by nitrite (0.5 mg·L-1 in terms of N), 46 metabolic compounds with significant differences were identified, involving amino acid metabolism, fatty acid metabolism, and glycerophospholipid metabolism. At pH=9.0, after stress (1.0 mg·L-1 in terms of N, i.e., non-ionic ammonia nitrogen was 0.378 mg·L-1), 52 significantly differential metabolic compounds were identified, involving tricarboxylic acid cycle, amino acid metabolism, fatty acid metabolism and glycerophospholipid metabolism. The quantitative test of the tricarboxylic acid cycle in the hepatopancreas of Macrobrachium nipponense was carried out in the pH=9.0 ammonia (1.0 mg·L-1) stress group. The results showed that the content of oxaloacetate in the hepatopancreas of Macrobrachium nipponense were significantly elevated at 12 h compared with the control group, but no significant differences were detected at 24 h and 48 h.α-ketoglutaric acid in the hepatopancreas of Macrobrachium nipponense were significantly elevated at 12 h compared with the control group, and decreased significantly at 24 h, but no significant differences were detected at 48 h. It may be concluded that the ammonia nitrogen (1.0 mg·L-1) stress lasting for 12 h at pH=9.0 will affect the tricarboxylic acid cycle.
点击查看大图
下载:
