The Clinical Study Of Type 2 Diabetic Nephropathy Ⅲ Stage With Qi And Yin Deficiency Syndrome Based On Metabonomics

ObjectiveTO identify the related biomarker of type 2 diabetic nephropathy(DN) Ⅲ stage with qi-yin deficiency syndrome based on LC/MS, and find out potential metabolism pathway in DN upon the different metabolic biomarkers.MethodsSerum samples of 30 DN Ⅲ stage patients,30 Type 2 diabetes mellitus(DM) patients and 30 healthy volunteers (NC) were collected. According to TCM theory, 30 patients of DN were qi-yin deficiency. The samples were detected with Liquid Chromatography-tandem mass spectrometry(LC/MS), and principal components analysis(PCA) and orthogonal partial least squares-discrimination analysis (OPLS-DA) were performed to find out different metabolites between them which were known as potential biomarkers. Then the potential biomarkers were identified by KEGG and HMDB to study the potential metabolism pathways and signaling pathways which may effect occurrence and development of DN.Results1. There was significant difference in age, course and body mass index(BMI) among DN group, DM group and NC group. There was no significant difference in TC, TG, LDL-C, Scr. But FBG, HbAlc, HDL-C, BUN, ALB, UA,24h-U-Alb were significant difference among the three groups.2. PCA scores plots appears:3 groups could obviously discriminate;DN group and NC group, DM group and DN group also could discriminate. The different biomarkers among the 3 groups were:lysophosphatidy lethanolamine (LysoPE), lysophosphatidylcholines(LysoPC),phosphatidylinositol(PI), phosphatidyleth anolamine(PE), phosphatidylglycerol(PG), phosphatidicacid, chenodeoxycholic acid, sphingomyelin(SM), cardiolipin, monoacylglyceride(MG), diglyceride (DG), triglycerides(TG), phosphatidylcholine(PC), phosphatidylserine(PS),5-h ydroxylysine, aspartyl-Alanine, N(6)-(Octanoyl)lysine, linoelaidic acid, lactosamine, ganglioside GA2.3. There were different metabolites between DN groups with qi-yin deficiency syndrome and NC group. Up regulated potential biomarkers were lysophosphatidylcholines(LysoPC), diglyceride(DG), triglycerides(TG), hyaluronic acid. Down regulated potential biomarkers were lysophsphatidicacid, phosphatidylcholine(PC), lysophosphatidylethanolamine(LysoPE), phosphatidyl ethanolamine(PE), phosphatidylinositol(PI), phosphatidylinositolphosphate(P IP), Phosphatidylglycerophosphate, phosphatidylglycerol(PG), sphingomyelin(S M), cardiolipin, phosphatidylserine(PS), ganglioside GA2, Crotonoyl-CoA, Pteroyl-D-glutamic acid, Adenosine diphosphate, Dihydroxyace tone phosphate.4. There were different metabolites between DN groups and DM group. Up regulated potential biomarkers were lysophosphatidy lethanolamine (LysoPE), lysophosphatidylcholines(LysoPC), phosphatidylcholine(PC), phosphatidylglyc erolphosphate,phosphatidic acid, monoacylglyceride(MG), triglycerides(TG), glutaminyl-arginine, galac tosylhydroxylysine, chenodeoxycholic acid, hyaluronic acid. Down regulated potential biomarkers were phosphatidylethanolamine(PE), phosphatidy linositolphosphate(PIP), phosphatidylglycerol(PG), phosphatidylserine (PS),s phingomyelin (SM), cardiolipin, diglyceride(DG), gangliosideGA2, phenylalanyl-alanine.5. Analyze the different bioinformatics of metabolites, and found out the probable classical metabolism pathways in both DN and DM were glycerolipid metabolism, choline metabolism, arachidonic acid metabolism, linolenic acid, fatty acid, lysine and tryptophan metabolism, sphingolipid metabolism, benzoic acid and aminobenzoic acid metabolism, glyoxylate metabolism, dicarboxylate and methyl butyrate metabolism, glucolysis and gluconeogenesis pathway, biosynthesis of glycosylation phosphatidyl inositol anchor, oxidative phosphorylation pathway, platelet activation. Compared with DM, the different metabolism pathways in DN were purine metabolis. The probable signaling pathways in both DN and DM group were cAMP signaling pathway、 Phosphatidylinositol signaling pathway, GnRH signaling pathway,FoxO signaling pathway, AMPK signaling pathway.,Conelusion1. The potential metabonomic biomarkers were identified based on LC/MS among DN, DM and NC group, PCA score plot had the trend to discriminate the three groups. Identified 21 biomarkers which related to the metabolism of Glucose, fat and amino acid.2. There had 17 different metabonomic biomarkers between the DN Ⅲ stage with qi-yin deficiency syndrome group and healthy volenteers which related to the metabolism of Glucose, fat and amino acid.3 biomarkers were up regulated and 17 biomarkers were down regulated.3. Metabolic pathways and signaling pathway analyzed by bioinformatics may be the important pathways in the onset process of type 2 diabetic nephropathy, and it deserved further research.