基于非靶向代谢组学技术的氨基脲胁迫雌性栉孔扇贝的毒性作用机制研究

Research on toxicity mechanism of semicarbazide on female Chlamys farreri by untargeted metabonomics

中文关键词:  栉孔扇贝  氨基脲  代谢组学  差异代谢物  通路

英文关键词:Chlamys farreri  semicarbazide  metabolomics  differential metabolite  pathway

中文摘要:

      目的 基于非靶向代谢组学技术分析暴露于50 mg/L氨基脲72 h的雌性栉孔扇贝性腺代谢物组成及含量变化, 以探究氨基脲对雌性栉孔扇贝性腺组织毒性及其作用机制。方法 采用基于液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry, LC-MS/MS)的非靶向代谢组学技术手段, 对氨基脲胁迫栉孔扇贝的性腺损伤和毒性机制进行了研究。结果 非靶向代谢组学结果表明: 与对照组相比, 在氨基脲胁迫下, 雌性栉孔扇贝性腺组织代谢物发生改变, 共有64种差异表达代谢物, 其中上调33个, 下调31个; 在HMDB数据库中进行比对分析, 注释到36个差异表达代谢物, 主要功能分类为有机酸及其衍生物、脂类和类脂分子及有机氮化合物等。差异代谢物功能通路的富集结果表明: 氨基酸代谢、脂质代谢、信号转导、核苷酸代谢和内分泌系统等生物过程受到显著影响。根据筛选出的差异代谢物的生理功能及其涉及的代谢通路分析, 差异代谢物鸟氨酸上调, 推断雌性栉孔扇贝通过增加鸟氨酸含量, 维持其正常生长过程。推测氨基脲可能通过影响栉孔扇贝的AMPKα-2 mRNA表达, 进而影响了核苷酸代谢通路和类脂肪代谢作用通路, 使得栉孔扇贝体内软脂酸含量下降。氨基脲可能通过对NMDAR的拮抗作用, 干扰神经信号传导, 导致雌性栉孔扇贝相关行为异常。结论 本研究探究了氨基脲对雌性栉孔扇贝性腺组织毒性作用及栉孔扇贝的响应调控机制, 为后期深入研究氨基脲毒性作用机制提供理论依据。

英文摘要:

      Objective To explore the toxicity of semicarbazide on the gonadal tissue of female Chlamys farreri and its mechanism of action by analyzing the composition and content changes of gonadal metabolites of female Chlamys farreri exposed to 50 mg/L semicarbazide for 72 hours based on the non-targeted metabonomics technology. Methods The mechanism of toxicity mechanism of semicarbazide on female Chlamys farreri was studied by non-targeted metabolomics technology based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results Untargeted metabolomics results showed that the metabolites in gill tissue of Chlamys farreri were changed under semicarbazide stress, and there were 64 kinds of differentially expressed metabolites compared with the control group, which 33 were up-regulated and 31 down-regulated. Comparative analysis was carried out in HMDB database, and 36 metabolites were noted, including organic acids and derivatives, lipids and lipid-like molecules, organic nitrogen compounds, etc.. The enrichment results of functional pathways of the metabolites above showed that biological processes were significantly affected, such as amino acid metabolism, lipid metabolism, signal transduction, nucleotide metabolism and endocrine system. According to the pathways and physiological functions involved of the differential metabolites selected, ornithine was up-regulated, and it was inferred that Chlamys farreri maintained its normal growth process by increasing its content. It was speculated that semicarbazide might affect the expression of AMPKα-2 mRNA, inducing the pathway of nucleotide metabolism and lipoid metabolism changed and the decrease of the content of palmitic acid. The transmission of the nerve signal might be interfered by antagonizing NMDAR, leading to the behavior abnormal. Conclusion This research explores the toxicity mechanism of semicarbazide on gill tissue and the response regulation mechanism of Chlamys farreri, and provides the theoretical basis for further research on the toxic mechanism of semicarbazide.