| In order to explore the mechanism of chronic toxicity of perfluorooctane sulfonate on aquatic animal and find potential markers related to chronic toxicity,GC-MS metabolomics method was used to study the effects of perfluorooctane sulfonate on Liver and muscle endogenous metabolites changes,while transcriptominc method was used to explore the molecular mechanism of perfluorooctane sulfonate exposure on Liver.1.In this experiment,there were four groups including control group,PFOS concentration at 10,100 and 250?g·L-1 group.Liver samples were obtained at day 10,20,30 and 40,respectively.The experimental results are as following:At day 10,The results showed that eight metabolites including Lactic acid,L-Alanine,Oxalic acid,L-Proline,Glycine,Pyroglutamic acid?L-Glutamic acid in zebrafish were identified from in the liver of zebrafish which PFOS concentration at 10?g·L-1 group changed significantly comparing to the control group,metabolic pathways including D-Glutamine and D-glutamate metabolism,Glycine,serine and threonine metabolism,Alanine,aspartate and glutamate metabolism,Arginine and proline metabolism,Pyruvate metabolism,Glutathione metabolism,Pentose phosphate pathway,Glycolysis or Gluconeogenesis were affected.Five metabolites including Lactic acid,L-Alanine,Oxalic acid,L-Proline,Pyroglutamic acid,Glutamic acid,L-Glutamine,were identified from in the liver of zebrafish which PFOS concentration at 100?g·L-1 group changed significantly comparing to the control group,metabolic pathways including D-Glutamine and D-glutamate,metabolism Alanine,aspartate and glutamate metabolism,Arginine and proline metabolism,Pyruvate metabolism,Glutathione metabolism were affected.Nine metabolites including Lactic acid,L-Alanine,Oxalic acid,L-Proline,Pyroglutamic acid,Glutamic acid,Glycerol-3-phosphate,Alpha-D-Glucopyranoside,Glucose6-phosphate,D-Ribose-5-phosphate,were identified from in the liver of zebrafish which PFOS concentration at 250?g·L-1 group changed significantly comparing to the control group,metabolic pathways including D-Glutamine and D-glutamate metabolism,Starch and sucrose metabolism,Pentose phosphate pathway,Alanine,aspartate and glutamate metabolism,Arginine and proline metabolism,Pyruvate metabolism,Amino sugar and nucleotide sugar metabolism,Glutathione metabolism,Glycolysis or Gluconeogenesis,Galactose metabolism,Purine metabolism were affected.At day 20,the results showed that in experimental groups of PFOS 10,100,and 250?g·L-1,two metabolites in zebrafish liver,including mannose and galactose,changed significantly comparing to the control group.Metabolic pathways including Galactose metabolism,Fructose and mannose metabolism were affected.At day 30,the results showed that in experimental groups of PFOS 10and 250?g·L-1,ethanolamine in zebrafish liver,changed significantly comparing to the control group.No metabolic pathways were affected;L-proline,in zebrafish were identified from in the liver of zebrafish which PFOS concentration at 100?g·L-1 changed significantly comparing to the control group.At day 40,The results showed that four metabolites including taurine,phosphoric acid amide,?-D-glucose and Oleic acid in zebrafish were identified in the liver of zebrafish which PFOS concentration at 10?g·L-1group changed significantly comparing to the control group,four metabolic pathways including Taurine and hypotaurine metabolism,Sphingolipid metabolism,Glycerophospholipid metabolism,and Glycolysis or Gluconeogenesis were affected.Five metabolites including taurine,?-D-glucose,palmitic acid,oleic acid and cholesterol,were identified from in the liver of zebrafish which PFOS concentration at 100?g·L-1 group changed significantly comparing to the control group,four metabolic pathways including Primary bile acid biosynthesis,Taurine and hypotaurine metabolism,Steroid biosynthesis,Steroid hormone biosynthesis and Glycolysis or Gluconeogenesis were affected.Four metabolitesincludingtaurine,?-D-glucose,lacticacid,glycerol-3-phosphate and phosphoacetamide,were identified from in the liver of zebrafish which PFOS concentration at 250?g·L-1 group changed significantly comparing to the control group,four metabolic pathways including Taurine and hypotaurine metabolism,Glycerophospholipid metabolism,Glycerolipid metabolism,Sphingolipid metabolism and Glycolysis or Gluconeogenesis were affected.2.In this experiment,there are four groups,control group,PFOS concentration at 10,100 and 250?g·L-1 group.muscle were sampled at 40d.The results showed that six metabolites including Glycine,L-Threonine,L-Aspartate,Taurine,L-Histidine,Glucose-6-phosphate in zebrafish were identified from in the liver of zebrafish which PFOS concentration at 10?g·L-1 group changed significantly comparing to the control group,metabolic pathways including Glycine,serine and threonine metabolism,Histidine metabolism,Taurine and hypotaurine metabolism,Alanine,aspartate and glutamate metabolism,Pentose phosphate pathway,Glycolysis or Gluconeogenesis,Glutathione metabolism.Nine metabolites including Ethanolamine,L-Leucine,Glycine,Pyroglutamic acid?L-Proline,Creatinine,D-Ribose-5-phosphate,Glucose-6-phosphate,Cholesterol,were identified from in the liver of zebrafish which PFOS concentration at 100?g·L-1 group changed significantly comparing to the control group,metabolic pathways including Valine,leucine and isoleucine biosynthesis,Glycine,serine and threonine metabolism,Pentose phosphate pathway,Primary bile acid biosynthesis,Arginine and proline metabolism,Steroid biosynthesis,Glutathione metabolism,Glycolysis or Gluconeogenesis,Purine me-tabolism,Steroid hormone biosynthesis were affected.Eight metabolites including Lactic acid,L-Proline,Creatinine,Glycerol3-phosphate,L-Histidine,D-Ribose-5-phosphate,Glucose 6-phosphate,Inosine,D-Ribose-5-phosphate,were identified from in the liver of zebrafish which PFOS concentration at 250?g·L-1 group changed significantly comparing to the control group,four metabolic pathways including Pentose phosphate pathway,Histidine metabolism,Pyruvate metabolism,Glycerophospholipid metabolism,Arginine and proline metabolism,Glycerolipid metabolism,Glycolysis or Gluconeogenesis,Purine metabolism were affected.3.After analyzing the metabolome data of the zebrafish on liver,the group having the most of different metabolites was selected,and the transcriptome was sequenced to explore the toxic mechanism of PFOS.The results showed:test group compared with the control group,under the FPKM analysis in the condition of fold change>2 and false discovery rate?FDR?<0.05,showed that the expression of 95 transcripts were significantly changed,61 genes were up-regulated and 34 genes were down-regulated;The GO enrichment analyses were performed on the functions of reference and new genes.The biological process-related pathways high enriched by the reference and new genes were:steroid metabolism,lipid metabolism,sterol metabolism,and sterol biosynthesis.Lipid biosynthetic processes and other processes,in addition to the same pathway,the new genes were also significant in the KEGG pathway for propionate metabolism,myocardial contraction,AMPK signaling pathway,sphingolipid metabolism influences.The experimental results are:1.The metabolomics study on the effect of PFOS on the liver showed that:In the short-term,high concentrations of PFOS have more significant impact on the liver,the three experimental groups affected amino acid metabolism,pyruvate metabolism pathways at the same time.In high concentration of PFOS group,glucose metabolism and purine metabolism were affect greatly;In the long-term,taurine and hypotaurine metabolism,glucose metabolism,lipid-related Metabolic were effects greater.2.Exposure to PFOS may cause muscle oxidative damage and cause an immune reaction.In addition,in the long-term,higher concentrations of PFOS will lead to creatinine increased significantly in muscle,which suggesting that PFOS can lead to kidney damage and increase the risk of kidney disease.3.Exploring the potential regulation of PFOS on the liver of zebrafish at the genetic level,the results showed that PFOS can effect greater on the lipid metabolism of hepatocyte genes,and PFOS impacted muscle composition and physiological functions significantly,by cell components and molecular function.PFOS potentially affects the myocardial contraction process by AMPK signaling pathway,and PFOS can effect on sphingolipid metabolism,it have potential toxic that effects on the zebrafish heart. |
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