Proteomics analysis of liver proteins from rats with spleen-deficiency syndrome induced by chronic improper diet consumption and fatigue

Objective:To investigate a proteomics analysis of liver proteins from rats with spleen-deficiency syndrome (SDS) induced by chronic improper diet consumption and fatigue.Methods:This study used a liver proteomic profiling method to identify differentially expressed proteins and altered pathways involved in SDS rats.Specifically,we collected liver samples from a control group and a group with SDS induced by chronic improper diet consumption and fatigue for 4 weeks.The pooled liver proteins in each group were labeled with 8-plex isobaric tags for relative and absolute quantitation reagents.The labeled control and SDS group samples were pooled together and separated by high-pH reverse-phase liquid chromatography.The differentially expressed proteins from the liver proteomes were analyzed to identify potential biomarkers of SDS.Differentially expressed proteins were selected in conjunction with gene ontology and ingenuity pathway analysis.Results:We identified 2176 protein clusters with more than two peptides in the SDS group,with 141 proteins quantified as differentially expressed proteins.Of these,75 proteins were up-regulated,and 66 were down-regulated.Three activated signaling pathways,the thrombin,CXCR4 and synaptic long term depression signaling pathways,and a large multi-protein complex within the network were revealed in the liver proteomic analysis of SDS rats.Conclusions:This is the first report of a differential liver proteome under SDS conditions.The results suggest that the liver proteome partially reflects the pathological changes involved in SDS.The findings provide important information for comprehensively understanding the mechanisms of dysfunction or injury in the liver at the molecular level as a result of SDS.Furthermore,they provide a novel understanding of the connotation of SDS in the fleld of traditional Chinese Medicine.

Gut microbiota and Chinese medicine syndrome: altered fecal microbiotas in spleen(Pi)-deficient patients

OBJECTIVE:To investigate the fecal microbiota changes in patients with spleen(Pi)-deficiency(SPD),a common Chinese medicine syndrome with digestive and absorptive disturbances and to provide insight into the relationship between Chinese medicine syndrome and gut dysbiosis.METHODS:Fecal microbiotas from the stool samples of 53 SPD patients and 35 healthy subjects were analyzed via 16 S r RNA gene polymerase chain reaction(PCR)-denaturation gradient gel electrophoresis(DGGE).SPD-related marker genes from20 SPD patients and 20 healthy subjects were identified through gene sequencing,while some genes were quantified using quantitative PCR(q PCR).Discriminant analysis was conducted using SPSS software,and the canonical discriminant function formula for Pi-deficiency was established.RESULTS:Alterations in microbiota diversity and composition between the SPD and healthy groups were demonstrated via 16 S r RNA gene PCR-DGGE combined with multivariate statistical analysis.Fecal microbiota changes were also observed among different SPD-subtype patients.Eight SPD-related markers were found,and putative species corresponding to these markers were identified through gene sequencing,which may have potential associations with the common digestive dysfunctions in SPD patients.q PCR methods were established for two of these markers,which were significantly altered in the SPD patients.The canonical discriminant function formula was calculated for SPD,and the validity rates of these markers were over 85%.CONCLUSION:Fecal microbiotas are altered in patients with SPD,which may provide insight for further studies on clinically diagnosing and treating SPD.The results may also provide data to gain a better understanding of Traditional Chinese Medicine syndrome and gut dysbiosis.

Differentially Expressed Proteins in Rat Hippocampus after Chronic Immobilization Stress and Intervention Using Xiao Yao San Decoction

Objective To identify differentially expressed proteins in the hippocampus of rats after chronic immobilization stress(CIS)using a proteomics approach,and to study the effect of the Xiao Yao San(XYS)decoction on differentially expressed proteins.Methods Twenty-four Sprague Dawley rats were randomly assigned to one of four groups of equal body weight:control(non-stress),7-day stress,21-day stress and21-day stress+XYS treatment groups.Two-dimensional gel electrophoresis(2-DE)was used to detect differences in protein expression in rat hippocampus.One differentially expressed protein was measured and verified by western blotting.Results Seventeen proteins showed differential expression.Among these,eight could be identified:glial fibrillary acidic protein-2(GFAP-2),tubulin alpha-1c,cytoplasmic muscle actin2,14-3-3protein,β-2a tubulin,phosphatidylethanolamine binding protein,synucleinαsyn3,and a low molecular weight(18kD)protein.Six of these proteins exhibited increased expression,one showed decreased expression,and the other protein,which comprised five subtypes,were either increased or decreased.These proteins are known to be involved in immunity,signal transduction,cell cycle control,apoptosis,regulation of enzyme activity,cytoskeleton structure,and synaptic plasticity.GFAP-2was further analyzed,and its differential expression confirmed by western blotting.Conclusion Some proteins are differentially expressed in the hippocampus of rats under chronic stress.The biological functions of these differentially expressed proteins are varied.Finally,the XYS decoction can significantly up-or down-regulate these protein expression levels.