The Research Of Proteome Of TCM Symptom In Early Diabetic Nephropathy

Objective To investigate the relation between TCM symptom in early diabetic nephropathy with BMI,UAER and plasma level of homocystien(HCY).To select the different protein among the patients with symptom of deficiency of Yin both liver and kidney,with symptom of deficiency of Qi and Yin and patients with symptom of deficiency of Yang both spleen and kidney.To find the biomarker of TCM symptoms of early diabetic nephropathy.MethodsChoose the early diabetic nephropathy patients(Mogensen stage Phase ?)who meet the standards,and after TCM syndrome differentiation,50 cases were selected in syndrome of yin deficiency of liver and kidney,syndrome of deficiency of qi and yin,and syndrome of Yang deficiency of spleen and kidney,respectively.Their height,weight,blood sugar,kidney function,urine routine,and 24-hour urinary albumin and serum homocysteine were tested and compared.Eight urine samples were randomly selected from the three types of syndromes,extract and concentrate the urine protein through desalination,ultra-filtration,and centrifugation,take urine bidirectional protein electrophoresis,gel map stained by silver nitrate was analyzed through software to look for different protein spots.The different proteins was undertaken MALDI-TOF-MS peptide mass fingerprint test after gel in situ enzymolysis,search the NCBI protein database through Mascot Distiller,comprehensively analyze the queried results combined with the isoelectric points,molecular mass,quantity and coverage rate of matched peptides in the corresponding points in the two-dimensional gel electrophoretogram,to find the matching protein,identify the protein name and the molecules,then randomly select 4 urine samples from each type of syndrome,conduct the Western-blot validation of dynein and transferring.Results(1)The UAER of the group with symptom of deficiency of Yin both liver and kidney,with symptom of deficiency of Qi and Yin and the group with symptom of deficiency of Yang both spleen and kidney is 96.24±46.63 mg/24h,93.38±39.14 mg/24h,101.32±149.65 mg/24h.There is no obvious difference from statistics(P>0.05).The plasma level of HCY in above group is 15.26±2.41 umol/1.20.38±2.69 umol/1?25.46±2.42 umol/1,they become gradually higher,there is significant difference from statistics(P<0.01).BMI in above group is 18.40±1.29 Kg/m2?26.72±2.12 Kg/m2?27.05±1.89 Kg/m2,the group with symptom of deficiency of Yin both liver and kidney is lower than other groups,there is obviously difference from statistics(P<0.01).(2)There are 789 protein spots,879 protein spots and 791 spots respectively in the group with symptom of deficiency of Yin both liver and kidney,with symptom of deficiency of Qi and Yin and patients with symptom of deficiency of Yang both spleen and kidney,found 22 different protein spots.(3)12 kinds of different protein matched highly were recognized by mass spectrometry.Their name is serum albumin,NOGO-interacting mitochondrial protein?Ig kappa chain,NIG93 precursor-human,Cytoplasmic dynein 1 heavy chain 1,transferring,Monoclonal 1gM antibody chain,Transthyretin,Prostate-specific antigen,Alpha-1-antitrypsin(aa 268-394),Chain A.There are 3 kinds serum human albumin decorated.(4)Dynein and transferring is demonstrated obviously in the group with symptom of deficiency of Qi and Yin than other groups by western-blot.ConclusionFirst,BMI is relevant to TCM symptom in early diabetic nephropathy.Second,plasma HCY could be acted as one of the proof of distinguishing TCM symptom in early diabetic nephropathy.Third,2DE-MS could be acted as the tool to select the urine different protein among TCM symptom.In the end,11 kinds of different protein such as dynein,Transthyretin and transferring could be the different protein of TCM symptom in early diabetic nephropathy...