| Objective:This project adopts a new proteomics technology(iTRAQ Technology),screening of different proteins in patients’ plasma with modified Buzhongyiqi Decoction in the treatment of myasthenia gravis,building the protein database,and making the correlation analysis of differentially expressed proteins in patients with clinical signs,symptoms and curative effect.Then we dig the target protein from drugs,which provide theoretical and experimental basis for evaluation of Traditional Chinese Medicine for treatment of myasthenia gravis.Methods:12 plasma samples which meet the inclusion criteria were collected,it were divided into four groups : normal control group(C,mixed labeled 113),untreated group(T1,mixed labeled 114),routine treatment group(T2,mixed labeled 115),routine treatment plus traditional Chinese medicine group(T3,mixed marking 116),3 cases in each group.The untreated group were not diagnosed for the first time as MG;the routine treatment group took pyridostigmine or pyridostigmine tablets + hormone therapy;the routine treatment plus traditional Chinese medicine group was based on the routine treatment,combined with Buzhong Yiqi Decoction in morning and evening between each time treatment,course of treatment is 2 a month.At the end of sampling,it were carried out about protein quality control,mass spectrometry analysis and biological information analysis on the samples in turn,and made the correlation analysis between the differential proteins and the efficacy.Results:In this project,a total of 618 proteins were identified(calculated according to three batches)of which have the quantitative results among 447 proteins(114:113,115:113,in 115:114,116:113,116:114,116:115),the number of significant differences proteins in the quantitative results respectively is 110,117,143,115,86 158,the number of up-regulated proteins respectively were 67,229,191,64,38,82,44,the number of down regulated proteins 252,242,51,48,76.T3 is better than group T2 in the clinical efficacy,and focused on the T2 and T3 group of the differential protein of the cell components of significant enrichment analysis,obtained 17 for different GO function enrichment entries of the differential protein(extracellular region,extracellular region part,extracellular organelles,etc.).The Pathway enrichment were made between T3 and T2 group for difference of protein,we found that the main metabolic pathway and signal transduction pathways is for the complement system(Dissolved target cell,promote phagocytosis,and neutralizing lytic virus,the role of inflammatory mediators),Fc gamma R mediated phagocytosis(leading to actin polymerization and phagosome formation)for up regulated proteins,metabolic pathway for down regulated proteins etc.Conclusion:1、The normal control group(C),untreated group(T1),routine treatment group(T2)and routine treatment plus traditional Chinese medicine group(T3).Four group of plasma proteome proteins which is for quantitative 、differenced proteins and functional annotation of protein have obvious differences,and comparison between every two groups,combined with clinical curative effect,T3 group is obvious effect.A total of 158 proteins for GO function enrichment analysis are obtained by comparison of T3 and T2,and it is closely related by the extracellular,high density lipoprotein particles and lipid protein complex,integrin,neuromuscular junction and extracellular space etc.Corresponding to database of(http://www.uniprot.org/),the 158 differential proteins(Protein AMBP,Integrin,alpha-IIb,etc.)are the target proteins of the drug action,which is the mechanism of modified Buzhongyiqi Decoction on the level of protein in the treatment of myasthenia gravis.2、Due to the limitations of iTRAQ technology,it were more difficult to collect samples,it is various reasons for the safety problem to compliance and clinical treatment after into groups of cases,without analysis of pure Chinese medicine group comparative statistics,and it still need further research. |
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