| Objective:To find differential metabolites and metabolic pathway to early diagnose iron deficiency anemia,metabolomic studies were conducted on urine of rural left-behind children under one year old with iron deficiency anemia in Zunyi area by hydrogen nuclear magnetic resonance testing technology.Methods:From October 2017 to February 2021,seven counties and districts under the jurisdiction of Zunyi area were selected as the study areas based on stratified sampling.Basic information,physical examination,dietary investigation and medical history inquiry were collected from children under 1 year old in the research areas.Blood and urine samples were collected.Routine blood tests and iron metabolism tests were performed on all the children.40 left-behind children diagnosed with iron deficiency anemia were randomly selected as the observation group,and 40 normal left-behind children were selected as the normal control group.The urine samples of the two groups were processed by hydrogen nuclear magnetic resonance detection technology to obtain ~1H-NMR spectra.Simca-p 13~+software was used to conduct principal component analysis,partial least square discriminant analysis and orthogonal partial least square discriminant analysis to determine whether there were differences in urine metabolites between the two groups and screen for differential metabolites.One-variate paired t-test was performed for the metabolites obtained,and metabolic pathways were finally analyzed by Metabo Analyst 5.0.Results:There was no significant difference in sex ratio,age,length and weight between the two groups(P>0.05).Normal control group serum ferritin was(47.82±16.04)ug/L,serum iron was(16.0±2.4)umol/L,hemoglobin was(123.9±4.4)g/L,Serum ferritin in the IDA group was(8.15±2.21)ug/L,serum iron was(8.4±2.0)umol/L,hemoglobin was(97.4±8.3)g/L,The differences in serum ferritin,serum iron and hemoglobin were significant between the two groups(P<0.0001).The ~1H-NMR spectra of urine of the two groups were obtained by nuclear magnetic resonance detection.The PCA model was established,and the integration value separation trend was obvious.Two principal components was obtained in this analysis:R2X=0.575,and Q2=0.369;The PLS-DA model was established and verified.The separation trend of integral values between the two groups was obvious.The model parameters R2Y=0.99,Q2=0.978,indicating that the model had not been fitted and the model intercept slope was large,the model was reliable.Further,the OPLS-DA model combined with ~1H-NMR spectra of metabolites in urine samples were used to obtain the score diagram and correlation coefficient load diagram.The model parameters R2X=0.345,R2Y=0.99,Q2=0.976,indicating that no fitting phenomenon occurred in the model.The model reliably found 5 substances with absolute correlation coefficients greater than the critical value of correlation coefficients.Univariate paired T-test was performed on the five metabolites obtained.The results showed isovalerylglycine,choline,phosphorylcholine/glycerophosphoric choline decreased and methylmalonate,2,3-dihydroxyisovalerate concentrations increased in the urine of children with iron-deficient anemia compared with normal controls(|r|>0.602,P<0.05).By Metabo Analyst 5.0 analysis,three metabolic pathways were identified,including valine,leucine,and isoleucine degradation,glycerophospholipid metabolism,glycine,serine,and threonine metabolism.Conclusion:There are differential metabolites between the urine of IDA children and normal children,and the production of these differential metabolites may be related to valine,leucine,and isoleucine degradation,glycerophospholipid metabolism,glycine,serine and threonine metabolism,and these changes can provide some clues for the early diagnosis of IDA. |
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