环境相关浓度睾酮暴露对秀丽隐杆线虫代谢谱的影响
Effects of Exposure to Environmentally Relevant Concentrations of Testosterone on Metabolic Profiles of Caenorhabditis elegans
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摘要: 睾酮主要通过禽畜粪便及处理不彻底的废水排放进入外环境,作为典型的内分泌干扰物之一,能够导致环境水体中鱼类雄性化,对环境健康及生态安全产生不利影响。将L1期秀丽隐杆线虫(Caenorhabditis elegans, C. elegans)暴露于环境相关浓度睾酮(0.1、1、10 μg·L-1)48 h,通过超高效液相色谱串联质谱(UPLC-MS/MS)技术检测初级代谢产物,观察生物体代谢活动的变化,筛选出环境相关浓度睾酮暴露后的差异代谢物及代谢通路以探讨其潜在毒性,并结合Lasso回归分析建立模型,筛选出在睾酮暴露秀丽隐杆线虫代谢扰动过程中发挥重要作用的物质。结果表明,环境相关浓度睾酮会导致秀丽隐杆线虫代谢谱的显著变化,引起519种代谢物显著升高和539种代谢物显著下降。对各暴露组显著差异代谢物筛选并对代谢通路进行富集分析,识别了相关代谢通路:鞘脂代谢、甘油磷脂代谢、色氨酸代谢、脂肪酸降解、泛酸与辅酶A的生物合成、半胱氨酸和蛋氨酸代谢、组氨酸代谢、精氨酸和脯氨酸代谢以及嘌呤代谢。其中,嘌呤、氨基酸及脂质代谢在高浓度睾酮暴露组扰动加剧,表明随着睾酮浓度升高,生殖功能受损更为严重。Lasso回归分析结果显示,甲酰基-N-乙酰基-5-甲氧基犬尿胺可能作为秀丽隐杆线虫受睾酮胁迫代谢异常过程中的潜在生物标志物。Abstract: Testosterone is mainly discharged into the external environment through livestock manure and incompletely treated wastewater. As one of the typical endocrine disruptors, testosterone can lead to the masculinization of fish in ambient water, and cause adverse effects on environmental health and ecological safety. The L1 larval of Caenorhabditis elegans (C. elegans) were exposed to environmentally relevant concentrations (0.1, 1 and 10 μg·L-1) of testosterone for 48 h. The primary metabolites were detected by ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to observe the changes in metabolic activities, screening out the differential metabolites and metabolic pathways and exploring the potential toxicity. Additionally, a model was established with Lasso regression analysis to screen out the metabolites that play an important role in the metabolic disturbance after testosterone exposure in C. elegans. Results showed that 519 metabolites were significantly increased and 539 metabolites were significantly decreased. Furthermore, those alteration of metabolites were analyzed by KEGG pathway enrichment, and main pathways was involved in sphingolipid metabolism, glycerol phospholipid metabolism, tryptophan metabolism, fatty acid degradation, pantothenic acid and coenzyme A biosynthesis, cysteine and methionine metabolism, histidine metabolism, arginine and proline metabolism, and purine metabolism. Among them, the disturbance of purine, amino acid and lipid metabolism was more obvious in 10 μg·L-1 exposure group, indicating that with the increase of testosterone concentration, the reproductive function was more seriously impaired. Lasso regression analysis showed that acetyl-N-formyl-5-methoxykynurenamine may be a potential biomarker in the disturbed metabolism of C. elegans under testosterone exposure.
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Key words:
- testosterone /
- Caenorhabditis elegans /
- metabonomics
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