Effects Of Fushengong Decoction On Rats With Chronic Renal Failure By Serum Proteomics

BackgroundChronic renal failure(CRF)has become a global health problem and bears a huge economic burden.Currently,for the treatment of CRF is mainly dialysis and renal transplantation,there is no available therapy to prevent the progress of renal failure.FuShengong Decoction(FSGD)is summarized by Professor of Chinese medicine master GuoZiguang,who add and deduct some herbs based on the classic formula ‘jisheng shenqi pills’ with 60 years of clinical experience.FSGD is composed of Radix astragali,Rehmannia glutinosa,Dioscorea opposita,Fructus Corni,Semen Plantaginis,Rdix Achyranthis Bidentatae,Cortex Moutan Radicis,Rhizoma Alismatis,Poria,Rhizoma Atractylodis,Cortex Eucommiae,Hirudo,Cortex Phellodendri.Mainly for non replacement therapy of early stage of CRF,FSGD can prevent renal irreversible change when get timely treatment at early stage of CRF.Our previous studies demonstrated that,the levels of BUN and SCr were decreased significantly,renal function improved significantly after the treatment of FSGD.However,the underlying molecular mechanisms of its effect remain unknown,further study is needed.Proteomics research is to take the quantitative and accurateidentification of the most protein in a complex mixture or a genome.Essentially refers to the characteristics of the protein on a large scale level,including the expression of protein,post-translational modification,protein and protein interaction,thus obtained comprehensive understanding about the expression of protein on the occurrence of disease and integrated cell metabolism.ObjectiveTo understand the underlying molecular mechanism and signaling pathway involved in the FSGD treatment of CRF and screen differentially expressed proteins in rats with CRF treated with FSGD.Further research the potential biomarkers of FSGD for the treatment of CRF.Methods33 male SD rats were grouped into control group,model group and FSGD group(11 in each group)at random.The control rats were fed normally,while the other two groups were fed with 0.5% adenine feed stuff to make the CRF model.After the models were successfully made,all rats received standard chow.Then,rats in control and model groups were given by gavage with intra-gastric normal saline(NS),while that in FSGD group received Fushengong decoction at the dose of 16 g/kg,once a day for 30 d.Differentially expressed proteins were screened by isobaric tags for relative and absolute quantitation(iTRAQ)coupled with nanoscale liquid chromatography tandem mass spectrometry(nanoHPLC-MS/MS),andthese identified proteins were later analyzed by gene ontology(GO)analysis,including molecular function,cellular component and biological process,the kyoto encyclopedia of genes and genomes(KEGG)and search tool for the retrieval of interacting genes/proteins(STRING).Additionally,haptoglobin(HP)and alpha-1-antitrypsin(AAT)were finally verified by enzyme-linked immunosorbent assays(ELISA),Western blot and real time PCR.ResultsA total of 417 proteins were identified.Nineteen differentially expressed proteins were identified in the FSGD group compared with the model group,of which 3 proteins were up-regulated,and 16 proteins were down-regulated.Cluster analysis indicated that inflammatory response was associated with these proteins and complement and coagulation cascade pathways were predominantly involved.The validation methods further confirmed that the levels of HP and AAT were significantly increased(P<0.05),while HP and AAT were significantly decreased after treatment of FSGD(P<0.05).ConclusionHP and AAT may be the important biomarkers in the pathogenesis of CRF and FSGD therapy.