| Objective:1.By detecting the changes of metabolic small molecules in bronchial alveolar lavage fluid of rats exposed to PM2.5 by nuclear magnetic resonance?NMR?, explore the mechanism of lung injury.2.By detecting the changes of metabolic small molecules in serum of rats exposed to PM2.5 by nuclear magnetic resonance?NMR?,and the cardiovascular injury indexes,explore the mechanism of cardiovascular injury.Method:A total of 36 healthy male SD rats weighing 180-200 g were randomly divided into control group?sterile saline?,PM2.5 low dose group?1.5 mg / kg · bw?,PM2.5 middle dose group?6mg / kg · bw?,PM2.5 high dose group(PM2.5 concentration of 24 ml / kg),PM2.5 water-soluble component group?24 mg / kg · bw?and PM2.5 water-insoluble group?24 mg / kg · bw?,PM2.5 high dose group?24 mg / kg · bw?.The rats were sacrificed by aerosol lung administration for 5 times.A total of 36 healthy male SD rats weighing 180-200 g were randomly divided into 6 groups: 0 day group,1 day group,3 day group,5 day group,7 day group and 9 day group Group 6,the next day exposure,the dose was 24 mg / kg · bw.After the end of the infection,the rats were sacrificed by the Uralan,and the abdominal aorta was collected and the right lung was washed with saline.The bronchoalveolar lavage fluid and serum were detected by nuclear magnetic resonance spectroscopy.The indexes of SOD,MDA,LDH and serum cardiovascular injury were detected by the kit.Using MestReNove software to analyze the spectrum,SMICA11.0 analysis of the integral value matrix to identify the differential metabolites,MetPA3.0 analysis of differential metabolites to find the target metabolic pathway,with SPSS13.0 on BALF SOD,MDA,LDH and differential metabolites.The single factor analysis of variance was also performed between the two groups.The LSD-t test was used to compare the differences between the metabolites and the cardiovascular damage in the serum.The test level was 0.05.Result:1.Study on the mechanism of lung injury induced by atmospheric PM2.5 in ratsPathological results: With the increase of dose and exposure time,the lungs will have lymphocyte infiltration,inflammatory exudation,alveolar cavity shrinkage finally closed,water-soluble component group lymphocyte infiltration,non-water soluble Group showed inflammatory exudation.LDH,MDA and SOD of BALF: LDH activity and MDA content of BALF increased with the increase of PM2.5 dose and prolonged exposure time,SOD activity decreased?P <0.05?.Compared with the control group,LDH and SOD activity and MDA content in the low dose group had no significant difference?P >0.05?,LDH activity in the middle dose group was significantly increased?P <0.05?,LDH activity and MDA?P <0.05?.There was no significant difference in LDH,SOD activity and MDA content between the 1 day group,the 3 day group,the 5 day group and the 7 day group?P <0.05?,but the activity of LDH and MDA in 9 days group were significantly increased and the activity of SOD was significantly decreased?P <0.05?.There was no significant difference in the content of PM2.5 water-soluble components between the two groups?P >0.05?,The content of MDA in PM2.5 non-water-soluble component group was significantly higher than that in control group?P <0.05?.Metabolomics analysis: low,medium and high dose groups were identified VIP> 1 differential metabolites 7 species,10 species and 11 species,of which low-dose group of different metabolites relative content compared with the control group no significant difference P >0.05).The relative contents of lactic acid,alanine,N-acetyl glycoprotein and O-acetyl glycoprotein in the middle dose group were significantly higher than those in the control group,the relative contents of choline,betaine,taurine and ?-glucose were significantly lower than those of control group?P <0.05?.The relative contents of alanine,N-acetyl glycoprotein,glycerophosphorylcholine and glycine in high dose group were significantly higher than those in control group?P <0.05?,the relative contents of betaine,taurine and ?-glucose were significantly lower than those of the control group?P <0.05?.1,3,5,7 and 9 days group were identified 10 different kinds of metabolites of VIP> 1,10 species,11 species,11 species and 11 species,The relative contents of alanine in 1 day group were compared with those of 0 day group?P <0.05?.The relative contents of lactic acid and alanine in 3 days group were significantly higher than those in the control group?P<0.05?.The relative contents of lactic acid,alanine,succinic acid,glycerophosphorylcholine and glycine in 5 days group were significantly higher than those in 0 day group?P <0.05?.The relative contents of lactic acid,alanine,N-acetyl glycoprotein,glyceryl succinate and glycine in 7 days group were significantly higher than those in 0 day group?P<0.05?.The relative contents of alanine,succinic acid,glycerophosphoryl-holine and glycine in the 9-day group were significantly higher than those in the 0-day group,and the relative contents of choline and betaine were significantly decreased?P <0.05?.Water-soluble and non-water-soluble components were identified 10 different kinds of metabolites of VIP> 1 and 11 species.The relative contents of choline,betaine,taurine and ?-glucose in water-soluble constituents were significantly lower than those in control group?P <0.05?.The relative content of alanine in the non-water-soluble component group was significantly higher than that in the control group?P <0.05?.From the enrichment analysis and topological analysis,we can see that the metabolic pathways with the response value greater than 0.1 are taurine and sub-taurine metabolism as well as glycine,serine and threonine metabolism,while the 1-day and 3-day groups only to taurine and sub-taurine metabolism,5,7 and 9 days group involved in taurine and sub-taurine metabolism and glycine,serine and threonine metabolism,water-soluble component groups also appear taurine and sub-taurine metabolism and glycine,serine and threonine metabolism,serine and threonine,and non-water-soluble groups only involve taurine and sub-taurine metabolism.2.Study on the Mechanism of Cardiovascular Injury in Rats Exposed to PM2.5Analysis of Serum Cardiovascular Injury Index: The levels of serum LDH and CKMB,MDA,BNP,CRP,ET-1 and TF were increased with the increase of exposure dose and prolonged exposure time?P <0.05?.The activities of LDH and CKMB and the contents of BNP and CRP in the middle dose group were significantly higher than those in the control group?P <0.05?.The LDH and CKMB activity and MDA in the high dose group were significantly higher than those in the control group?P <0.05?,and the contents of CKMB and TF were significantly higher than those of low dose group and middle dose group?P <0.05?,and the contents of BNP,CRP,ET-1 and TF were significantly increased?P <0.05?.The serum levels of BNP and CKMB in the 3-day group were significantly higher than those in the control group?P <0.05?.The LDH and CKMB in the 5-day group were significantly higher than those in the control group?P <0.05?.The contents of LDH and CKMB,BNP,ET-1 and TF were significantly increased in the 7th day group?P <0.05?,and the contents of CKMB and BNP were significantly higher than those in the 1 day group?P <0.05?.The activity of LDH and CKMB,MDA,BNP,ET-1 and TF in 9 days group were significantly higher than those in 0 day group and 1 day group?P <0.05?,and CKMB activity And TF levels were significantly higher than those in the 3-day and 5-day groups?P <0.05?.The content of LDH and the content of BNP in water-soluble component group were significantly increased?P <0.05?.Metabolomics analysis: The low,medium and high dose groups were able to identify 10,10 and 11 differentially metabolites of VIP> 1?There was no significant difference in the relative contents of different metabolites in the low dose group compared with the control group?P >0.05?.The relative content of triglyceride in the middle dose group was significantly higher than that in the control group?P <0.05?.The relative contents of ?-hydroxybutyric acid,acetone and triglyceride in high dose group were significantly higher than those in control group?P <0.05?.The relative contents of taurine,glutathione,betaine and ?-glucose were compared with those in control group Significantly decreased?P <0.05?.In the 1,3,5,7,and 9 days groups were identified the differential metabolites 11,10,10,13 and 12 of VIP>1 were identified.Compared with the 0 day group,Compared with the 0 day group,the relative contents of different metabolites in the 1 day group and the 3 day group were not significantly different from those in the 0 day group?P>0.05?.The relative content of taurine and betaine in 5 days group was significantly lower than that in 0 day group?P<0.05?.The relative contents of taurine,glutathione,betaine and ?-glucose in the 7-day group were significantly lower than those of 0-day group?P<0.05?,and the relative contents of ?-hydroxybutyric acid and acetone were significantly increased?P<0.05?.The relative contents of ?-hydroxybutyric acid,acetone and triglyceride in the 9-day group were significantly higher than those in the 0-day group?P<0.05?,and the relative contents of taurine,glutathione,betaine and ?-glucose were significantly lower than those of 0-day group?P<0.05?.The difference between the water-soluble component group and the non-water-soluble component group was 10 and 13,and the relative contents of the different metabolites in the water-soluble and non-water-soluble groups were not significantly different from those in the control group differences?P> 0.05?.Conclusion:1.PM2.5 exposure can cause lung injury in rats,bronchial alveolar filling small molecule metabolic profile changes,causing taurine and sub-taurine metabolic pathways and glycine,serine and threonine metabolic pathway disorders.May be related to the decrease in the relative content of choline,betaine,taurine and the relative increase in glycine content;water-soluble components may cause taurine and sub-taurine metabolic pathways as well as glycine,serine and threonine metabolism Pathway disorders,non-water-soluble components may cause disruption of glycine,serine and threonine metabolic pathways.2.PM2.5 can cause oxidative damage,inflammatory response,endothelial dysfunction,coagulation dysfunction,cardiac function and cardiomyocyte injury,damage of cardiovascular system and lead to small molecule metabolic profile in serum altered,causing taurine and sub-taurine metabolic pathways,glycerol metabolic pathways,glutathione metabolic pathway disorders,may be associated with taurine,glutathione,betaine relative reduction in content and triglycerides relative rise in content.And the effect of water-soluble and non-water-soluble components on metabolic pathways was not significant. |
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