Metabonomic Study On Lung Injury Induced By Paraquat Poisoning In Rats

Objective:To explore the metabolic changes of lung tissue and BALF in rats poisoned by paraquat(paraquat,PQ),to clarify the mechanism of lung injury induced by PQ poisoning from the level of metabonomics,and to find therapeutic targets and biomarkers of PQ poisoning.Methods: The SD rats were randomly divided into NC group(control group),paraquat-treated 14-day group and paraquat-treated 28-day group.The animal model of PQ poisoning was established by intragastric administration of 20% paraquat aqueous solution to 1 mL at one time according to the dose of 45mg/kg.Bronchoalveolar lavage fluid samples and lung tissue samples were collected at 14 days and 28 days after exposure,respectively.Lung tissue samples from the left lower lung were taken.The Lung injury and the level of pulmonary fibrosis were detected by HE staining,MASSON staining and Western blotting.After the metabonomic pretreatment of bronchoalveolar lavage fluid and lung tissue samples,UPLC-TOF-MS/MS metabonomics technology was used to detect the samples.MetaboScape3.0,a software provided by UPLCTOF-MS/MS instrument,was used to extract metabonomic data and identify metabolites.PCA,OPLS-DA,T test,cluster analysis,Metabolic profile analysis,differential metabolites screening and metabolic pathway were analyzed by Metaboanalyst 4.0 and SIMC-P14.1.Results: Compared with the control group,the lungs of the rats in the14-day group were predominantly inflammatory,with slight fibrosis;the fibrosis in the 28-day group was further aggravated.IL-6,Fibronectin,Collagen and ?-SMA were up-regulated in the lung tissues of the 14-day and 28-day groups.PCA and OPLS-DA analysis showed significant differences in the metabolic profiles of lung tissue and alveolar lavage fluid in the NC group(control group),the 14-day paraquat group,and the 28-day paraquat group.Compared with the control group,with the criteria of FC>1.5,P<0.05,VIP>1,Pantothenic acid,Proline betaine,Hydroxybutyrylcarnitine,8-Hydroxy-7-methylguanine,Fumaric acid,Methylmalonic acid,L-2-Hydroxyglutaric acid,Stearic acid,Palmitic acid,Glyceraldehyde,Citraconic acid,alpha-Ketoisovaleric acid,Monic acid,LysoPC(15:0),LCystathionine,Hydroxykynurenine was screened out in the lung tissue of the 14-day group and the 28-day group.ROC analysis suggested that 8-Hydroxy-7-methylguanine,14-HDoHE,beta-Alanine,Methylmalonic acid,Palmitic acid,Pantothenic acid,Proline betaine and Stearic acid can be used as diagnostic PQ induction Biomarkers of lung injury.Pathway analysis showed that amino acid metabolism,glucose metabolism,fat metabolism,and energy metabolism were related to PQ-induced lung injury.Conclusion: PQ poisoning-induced lung injury in rats is mainly inflammatory response and fibrosis,and its metabolic profile changes significantly.Amino acid metabolism,fat metabolism,glucose metabolism,energy metabolism and other metabolisms are involved in the pathophysiology of lung injury induced by PQ poisoning in rats.process.