A Study On The Mechanism Of "Qian-Jin-Wen-Wu Decoction" Intervenes Diabetic Kidney Disease Based On Proteomics And Metabolomics

Objective:Using proteomics and metabolomics techniques,the intervention effect of the Chinese medicine "Qian-Jin-Wen-Wu Decoction"(QWD)on rat diabetic kidney disease(DKD)model were investigated,and the mechanism and target of QWD treatment of DKD were preliminarily elucidated.Method:1.The rat model of streptozotocin DKD was established.QWD was administered to model animals,observing body characteristics and measuring biochemical indicators to evaluate main pharmacodynamic effects.2.The protein in rat kidney tissue was extracted by urea method,after iTRAQ-labeled,the sample was placed into a liquid chromatography-mass spectrometer(LC-MS),generating a mass spectrometric detection original file;Seted three comparison groups of B-vs-M,B-vs-BD and M-vs-MD,the raw data obtained by mass spectrometry is searched and analyzed by bioinformatics.Screened for reliable peptides and proteins;statistical analysis,identified differential proteins,performed differential protein cluster analysis,and sample correlation analysis;did GO,KEGG function annotation and enrichment analysis based on the corresponding species function database,and did differential proteins interaction analysis.3.Taken and pre-treated rat kidney tissue,the metabolites were detected in positive ion and negative ion modes respectively by LC-MS.Three comparison groups of B-vs-M,B-vs-BD and M-vs-MD were set to qualitatively and quantitatively analyze metabolites;data quality control,data normalization,univariate statistical analysis,multivariate statistical analysis and orthogonal partial least squares(OPLS-DA)analysis has been done.Metabolic pathway annotation and enrichment analysis of differential metabolites had been doing.4.Conjoint analysis of proteomics and metabolomics experiments has been done for finding pairs of relationships that are consistent with enzyme-metabolite linkages and building networks.Results:1.After administration,the general indexes and biochemical indexes of rats were different,and the data we got were statistically significant.2.Proteomics experiments showed that B-vs-BD group found 4 significantly up-regulated proteins and 58 significantly down-regulated proteins;in B-vs-M group,21 proteins were significantly up-regulated,and 26 proteins were down-regulated significantly.For M-vs-MD group,5 proteins were significantly up-regulated and 14 proteins were down-regulated significantly.GO and Pathway's functional annotation and enrichment analysis of differential proteins indicate that QWD affects the Metabolic processes such as biosynthesis and metabolism,amino acid metabolism,lipid metabolism,energy metabolism and carbohydrates metabolism by regulating the nervous system,excretory system,digestive system,and endocrine system to have a therapeutic effect on DKD.The results of differential protein interaction network showed that Krt10,Krt8,Krt19,etc.can be used as target proteins for the treatment of DKD by QWD;The biological mechanism of DKD pathogenesis is related to the cytoplasmic oxidative phosphorylation pathway.3.Metabolomics experiments showed that the total number of compounds detectable in positive ion mode was 1666,1137 were screened,and the number of compounds detectable in negative ion mode was 1803,789 were retained by screening;The metabolic pathway annotation and enrichment analysis of differential metabolites showed that QWD can regulate alpha linolenic acid and linoleic acid metabolism,glycerolipid metabolism,glycolysis,starch and sucrose metabolism,fructose and mannose degradation,gluconeogenesis,Warburg Effects,and it can also be involved in glycine and serine metabolism as well as purine metabolism.4.After the protein data were homologous to humans,B-vs-BD group,B-vs-M group and M-vs-MD group each found an enzyme-metabolite co-expression relationship of 1 pair,6 pairs and 2 pairs,among which the same Metabolites testosterone appeared in the B-vs-M group and the M-vs-MD group,and the same enzyme ALDH1B1 in the B-vs-M group and the B-vs-BD group.These results suggest that the molecular mechanism of QWD intervention in DKD may be to restore the mitochondrial oxidative phosphorylation dysfunction and enhance multiple metabolic pathways in the body,thereby repairing renal tissue to restore cell function.Conclusion:This paper successfully established an animal model of QWD intervention DKD,preliminarily clarified the mechanism of how QWD is regulating and repairing the renal tissue of diabetic nephropathy.The research results provide the experimental basis and research ideas for the follow-up drug-making research of QWD.