| Objective:Based on the metabolomics /proteomics research methods,3-month-old mice were used as controls to study the changes in the metabolic profiles and markers of plasma,spleen and kidney tissues in 20-month-old mice,combined with the expression of differential proteins in liver mitochondrial samples,to analyze the aging-related metabolic/protein markers,biological pathways,and the intervention of Liuwei Dihuang Pill to explain the aging mechanism and the biological basis of the regulation of Liuwei Dihuang Pill;It lays the foundation for the study of the relationship between prescriptions and syndromes.Method:On the basis of the previous research of the research group,3-months-old mice were selected as control group,and 20-month-old natural aging mice were randomly divided into model group and drug-administered group,20 mice in each group(10 males and 10 females).The mice in the drug-administered group were given the Liuwei Dihuang Pill solution;the control group and the model group were intragastrically administered with the same volume of normal saline.After continuous gavage for 30 days,plasma,spleen,and kidney tissue samples were prepared for metabolomics analysis;liver mitochondrial samples were prepared for proteomic analysis.Result:1.From the results of principal component analysis(PCA),it can be seen that the metabolic profiles of the three biological samples in the mice in the drug-administered group were far away from the model group in the three-dimensional space and were recalled to the control group.2.Compared with the control group,the plasma sample model group identified metabolic markers related to aging,including 22 male markers and20 female markers;Associated with 6 metabolic pathways.Liuwei Dihuang Pill can significantly interfere with 11 of the identified male markers and 6 ofthe identified female markers;Liuwei Dihuang Pill has a regulatory effect on4 of the 6 identified aging-related pathways,including Selenoamino acid metabolism;Arginine and proline metabolism;Steroid biosynthesis;Pyrimidine metabolism.3.Compared with the control group,the spleen tissue sample model group identified metabolic markers related to aging,including 43 male markers and 43 female markers;Associated with 15 metabolic pathways.Liuwei Dihuang Pill can significantly interfere with 40 of the identified male markers and 23 of the identified female markers;Liuwei Dihuang Pill has a regulatory effect on 13 of the 15 identified aging-related pathways.4.Compared with the control group,the kidney tissue sample model group identified metabolic markers related to aging,including 46 male markers and 40 female markers;Associated with 9 metabolic pathways.Liuwei Dihuang Pill can significantly interfere with 17 of the identified male markers and 12 of the identified female markers;Liuwei Dihuang Pill has a regulatory effect on 6 of the 9 identified aging-related pathways,including D-Glutamine and D-glutamate metabolism;Arachidonic acid metabolism;Biotin metabolism;Taurine and hypotaurine metabolism;Alanine,aspartate and glutamate metabolism;Pentose and glucuronate interconversions.5.By comparing and analyzing the differences in protein markers of liver mitochondrial samples between model group and control group mice,102 male aging-related protein markers were screened;108 female aging-related protein markers were screened.Related to 5 biological pathways.Liuwei Dihuang Pill can significantly regulate 90 of the identified male markers and 94 of the identified female markers.Liuwei Dihuang Pill has a regulatory effect on the 5 identified aging-related pathways,including m TOR signaling pathway;Alzheimer’s disease;Arachidonic acid metabolism;ECM receptor interaction;Glutathione metabolism.6.A systematic and comprehensive analysis of the results of metabolomics and proteomics found that the same pathways affected byLiuwei Dihuang Pill in two histological studies were Arachidonic acid metabolism and Glutathione metabolism.Conclusion:1.Compared with 3-month-old mice,the metabolic markers of plasma,spleen and kidney tissue samples of 20-month-old mice were significantly different,which were related to 6,15 and 9 aging-related metabolic pathways.2.Liuwei Dihuang Pill has obvious reversal effect on the metabolic profile and markers of plasma,spleen and kidney tissue samples of20-month-old mice.It has a regulatory effect on the 4,13,and 6 aging-related metabolic pathways that have been identified.3.Compared with 3-months-old mice,there were significant differences in liver mitochondrial sample protein markers in 20-month-old mice,which were associated with 5 biological pathways.Liuwei Dihuang Pill can significantly recall the protein markers and regulate the 5 aging-related biological pathways.4.Combined with the results of metabolomics and proteomics,a comprehensive analysis of system biology revealed that the same pathways affected by Liuwei Dihuang Pill in two omics studies were Arachidonic acid metabolism and Glutathione metabolism. |
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