| Objective:By detecting changes in serum and urine metabonomics of rats which exposured ambient PM2.5 for short-term, analysis the impact on possible metabolic pathways and search for the possible early biological markers to further exploring for the toxic mechanism which ambient PM2.5 damage to the body, and then provide a new theoretical basis for the effective prevention and controlling environmental pollution which is caused by fine particles damage to the human body. Method:Ambient PM2.5 particles was sampled by a large flow of air particulate sampler(Model: TH-1000CⅡ) at large traffic junctions in the traffic flow during heating. 36 healthy male SD rats were randomly divided into three dosage groups(1.5, 6, 24mg/kg body weight) and two component groups(water soluble, non-water soluble, 24 mg/kg body weight) and one saline control group of six rats each. Each group received aerosol exposuring one another day for a total of five times. Another 36 healthy male SD rats were randomly divided into five aging groups(1st, 2nd, 3rd, 4th, 5th, 24mg/kg body weight)and one saline control group. Expose the ne xt day by using rat aerosol pulmonary delivery suite, and five aging groups were exposed to 1,2,3,4,5 times respectively. Collect the urine of rats after exposing 24 hours, and blood in abdominal aortic after urethane anaesthetizing. Histopathological changes of heart, liver, spleen, lung, kidney were observed after embedded in paraffin, sectioned, stained. The metabolomics of serum and urine were detected by nuclear magnetic resonance(1H NMR, BRUKER AVANCEⅢ). Analysis the matrix by Mest ReNova and SMIC A P11.1, then find the different metabolites. The metabolic pathway metabolic differences were analyzed by HMDB(Human Metabolome Database) and MetPA(Metabolomics Pathway Analyst) database. Result:1. Pathological changesHistopathology showed, low dose group rats had alveolar collapse, and middle and high dose group, the group of water-soluble component, a water- insoluble component group showed pulmonary interstitial infiltration of lymphocytes, alveolar septa greatly increasing, inflammatory exudating. High-dose group, the group of water-soluble component, a water- insoluble component group appears alveolar closure. Rats myocardial hypertrophy were seen in low, medium and high dose groups and water-soluble group. Myocardial cells shrinkage were observed in the non-water-soluble. Liver periportal infiltration of lymphocytes was seen in low dose group. Hepatic steatosis were observed in medium and high dose groups, water-soluble components and non-water soluble ingredients. Inflammatory exudates were showed in middle dose group and high dose group. Tubular dilatation was seen in middle dose group. Full glomeruli was observed in high dose group. Glomerular capsule space was disappeared in the group of water-soluble components. Spleenin each group were normal.Pulmonary interstitial inflammatory cell infiltration and inflammatory exudate were seen in the group of PM2.5 exposed twice or more. Four times more exposed showed alveolar collapse. PM2.5 exposure more than three times had myocardial hypertrophy, liver periportal lymphocyte infiltration, hepatic steatosis, liver poisoning. PM2.5 exposure five times had inflammatory exudates and renal tubular dilation. Spleen in each group were normal.2. Metabolic changes on serum of ambient PM2.5 exposureCompared with the control group, the relative amount of dimethyl glycine in low-dose group was significantly higher, and the relative content of 3-hydroxybutyric acid, O-acetyl glycoproteins, creatine, creatinine decreased significantly, but there was no significant difference in the associated metabolic results(Impact<0.10); the relative content of L- lactic acid, N-acetyl glycoproteins were significantly higher in the median dose group, the content of 3- hydroxybutyric acid, glycerol phosphate, choline, beet base, taurine, glucose-6-phosphate were relatively decreased significantly, mainly involving taurine and hypotaurine metabolism, valine, leucine and isoleucine biosynthesis; in the group of large high-dose, the relative content of glycerol, isoleucine, dimethyl glycine, D-lactic acid increased significantly, and 3-hydroxybutyric acid, glycerol phosphate, choline, betaine, taurine, glucose-6-phosphate, D- glucose, proline betaine, triglycerides, α-D-1,6-diphosphate glucose were significantly decreased, and was mainly related to taurine and hypotaurine metabolism, valine, leucine and isoleucine biosynthesis, glycerol metabolic. In the group of water-soluble components, the relative content of L- methionine, isoleucine, L- lactic acid, glycerol phosphate, choline were significantly increased, and the relative content of dimethyl glycine, creatinine, D-glucose, betaine, lysine, creatine were decreased significantly, and was mainly related to valine, leucine and isoleucine biosynthesis pathway. The relative content of glycero l, isoleucine, dimethyl glycine were significantly increased, and L- lactic acid, D- glucose, betaine, taurocholic acid, glucose-6-phosphate, lysine acid, α-D-1,6-diphosphate glucose were decreased significantly in water-insoluble group, and was mainly related to valine, leucine and isoleucine biosynthesis pathways glycerides.Aging study results showed that compared with the control group, in the group of 1st, the relative content of 3-hydroxybutyric acid, acetoacetic acid, taurine, glucose-6-phosphate, triglycerides increased significantly, isoleucine acid, dimethyl glycine, D- lactic acid were relatively decreased significantly; in the group of 2nd, the relative content of 3-hydroxybutyric acid, glycerol phosphate, choline, TMAO, betaine, taurine, glucose-6-phosphate, glycerol-3-phosphoric acid, D- leucine, L- lysine, D-glucose, proline betaine, α-D-1,6- diphosphate glucose were significantly increased, and the relative content of L- lactic acid, N-acetyl glycoprotein, glycoprotein O-acetyl, dimethyl glycine, D-lactic acid were decreased significantly; in the 3rd group, the relative content of L- methionine, choline glycerophosphate, taurine, glucose-6-phosphate, glycine, D-leucyl acid, D- glucose, proline betaine, α-D-1,6- diphosphate glucose were significantly increased, the relative content of L-lactic acid, isoleucine, dimethylglycine, D-lactic acid decreased significantly;in the group of 4th, the relative content of L- methionine, 3-hydroxybutyric acid, acetoacetic acid increased significantly, the relative content of L- lactic acid, isoleucine, dimethyl glycine, D- lactic acid significantly decrease; in the group of 5th, the relative content of glycerol, isoleucine, dimethyl glycine, D-lactic acid increased significantly, the relative content of 3-hydroxybutyric acid, glycerol phosphate, choline, trimethylamine oxide, betaine, cattle acid, glucose-6-phosphate, D-glucose, proline betaine, triglyceride,α-D-1,6-diphosphate glucose significantly reduced. Changes above were associated with taurine and hypotaurine metabolism, valine, leucine and isoleucine metabolism. In addition, 1st group also relates synthesis and degradation of ketone bodies, butyrate Metabolism; 2nd, 3rd groups also related glycine, serine and threonine metabolic; 4th, 5th groups also related glycerol metabolism.3. Metabolic changes on urine of ambient PM2.5 exposureCompared with the control group,in the low dose group, the relative content of ethyl acetate, creatine, D- glucose, taurocholic acid, 5-hydroxy group increased significantly, the relative content of hippurate, creatinine, a maleimide uric acid was significantly lower, but not statistically significant(Impact <0.10) results related metabolic pathway. In the median dose group, the relative content of betaine, TC A were significantly increased, the relative content of citrate, α-ketoglutarate, hippurate, creatinine, methyl hippuric acid significantly reduced, mainly involving glyoxylic acid metabolism and the citric acid cycle. In the high dose group the relative content of muscle acid, taurocholic acid, L-prolyl- L-glycine significantly increased, the relative content of allantoin, citrate, dimethyl tryptamine, α-ketoglutarate, hippurate, creatinine methyl hippuric acid significantly reduced, mainly involving glyoxylic acid, citric acid metabolism and circulation. In the water soluble component group, the relative content of betaine, taurocholic acid, L- lysine, allantoin increased significantly, the relative content of α-ketoglutarate, citric acid, dimethylamine, taurine, hippurate, creatinine, methyl hippuric acid significantly reduced, mainly involving taurine and hypotaurine metabolism, metabolism of glyoxylic acid and a dicarboxylic acid, citric acid cycle. In the non-water soluble component group, the relative content of taurocholic acid, L-lysine significantly increased, relative content of citric acid, dimethylamine, hippurate, creatinine, methyl hippuric acid significantly reduced, mainly involving glyoxylic acid and dicarboxylic acid metabolism.Aging study showed, compared with the control group, in the 1st group, the relative content of betaine, taurocholic acid, 5- hydroxy, dimethyl glycine, L-lysine, guanidinoacetate increased, the relative content of ethyl acetate, succinate,α- ketoglutarate is significantly lower, but not statistically significant(Impact<0.10) results related metabolic pathways; in the group of 2nd, the relative content of taurocholic acid, glycylglycine, diethanolamine, 5-hydroxy, dimethyl glycine, L- lysine, guanidine acetic acid, uridine diphosphate glucose, hippurate, allantoin was significantly increased, the relative content of ethyl, creatine, creatinine decreased significantly, mainly involving pentose and glucuronate interconversions, starch and sucrose metabolism; in 3rd group, the relative content of betaine, taurocholic acid, 5-hydroxy, L- lysine significantly increased, and the relative content of succinic acid, creatinine decreased significantly, but the metabolic pathway analysis was not statistically significant(Impact <0.10); in the group of 4th, the relative content of L- lactic acid, pyruvic acid, taurocholic acid, 5-hydroxy, guanidine acetic acid increased significantly, the relative content of succinic acid, creatine, hippurate, creatinine, methyl hippuric acid significantly reduced, but the metabolic pathway analysis was not statistically significant(Impact<0.10); in the group of 5th, the relative content of taurocholic acid, L-prolyl-L- glycine, allantoin increased significantly, the relative content of citrate, dimethyl tryptamine, α-ketoglutarate acid, hippurate, creatinine, methyl hippuric acid decreased significantly, and mainly related to metabolism of glyoxylic acid and a dicarboxylic acid, citric acid cycle related. Conclusion:By dose-response relationship and the aging study, concluded that the ambient PM2.5 can lead to taurine, glucose-6-phosphate, D- glucose, glycerol phosphate, choline, betaine proline significantly reduced in the serum, and there is dose-response and time-dependent on the metabolic pathways. It mainly affected: taurine and hypotaurine metabolism, valine, leucine and isoleucine biosynthesis, glycerol metabolism, synthesis and degradation of ketone bodies, butanoate metabolism, glycine, serine and threonine metabolism.The atmospheric PM2.5 can lead to a significant decrease in rat urine hippurate, creatinine, uric acid methyl maleimide, and relyed on a dose-response and time-dependent. The major metabolic pathways are: glyoxylate and dicarboxylate metabolism, citrate cycle, taurine and hypotaurine metabolism, pentose and glucuronate interconversions, starch and sucrose metabolism.This topic is " Shanxi characteristics of air pollution and health damage prevention and control" of advantages and characteristics of Shanxi Key Discipline Construction Project. |
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