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.

Effect of Weiliuan of different prescriptions on gastrointestinal regulation factor in spleen-deficiency rats

OBJECTIVE To observe the regulating effect of Weiliuan on digestive enzyme and relevant gastrointestinal hormone in spleen-deficiency rats.METHODS The rats were given the decoction of Radix et Rhizoma combine with dietary disorder to induce spleen-deficiency syndrome.Spleen-deficiency rats were divided into 8groups:normal control group,spleen deficiency model group,Mengtuoshi San group(0.15g·kg-1),Weiliuan 31.5and 15.75g·kg-1 groups,Weiliuan splited prescriptionⅠ 31.5 and 15.75g·kg-1groups,Weiliuan splited prescriptionⅡ 31.5and 15.75g·kg-1 groups.At the end of experiment,the level of amylase(AMS),lactic dehydrogenase(LDH),succinate dehydrogenase(SDH),motilin(MTL),somatostatin(SS),gastrin(GAS)and organ coefficient were detected.RESULTS Weiliuan group,Weiliuan splited prescriptionⅠ group and Weiliuan splited prescriptionⅡ group can significantly reduce the spleen index,the level of SDH and LDH,it can also improve the level of the AMS(P<0.05,P<0.01)In the regulation of hormone levels,Weiliuan group,Weiliuan splited prescriptionⅠ group and Weiliuan splited prescriptionⅡ group can significantly improve the levels MTL and GAS(P<0.05),but had no obvious effect on SS.CONCLUSION Weiliuan group,Weiliuan splited prescriptionⅠ group and Weiliuan splited prescriptionⅡ group can improve the symptoms of spleen deficiency syndrome significantly.Its mechanism may be to regulate the levels of AMS,SDH,LDH,MTL and GAS.

Differential expression of immune-related genes between healthy volunteers and type 2 diabetic patients with spleen-deficiency pattern

OBJECTIVE: To investigate the clinical differentiation of spleen-deficiency pattern(SDP), a group of symptoms and signs defined in terms of Traditional Chinese Medicine for its clinical practice.METHODS: Peripheral venous blood(> 3 m L) was collected from each of six type 2 diabetes mellitus(T2DM)-SDP patients and six healthy volunteers. After the isolation of peripheral white blood cells(PWBCs), total RNA was extracted, and quality control was performed on all RNA samples. Microarray experiments were conducted using the Agilent human whole genome gene chip, and genes demonstrating differential expression were screened. Bioinformatics analysis was conducted on these genes using several online databases.RESULTS: We screened a total of 175 differentially expressed genes(DEGs), of which 111(63%) were down-regulated and 64(37%) were up-regulated in T2DM-SDP patients compared with healthy controls. Among the 175 genes, 158 had biological function annotations: 46(29%) were directly related to an individual's immune regulation or response, 25(16%) were associated with substance and energy metabolism of PWBCs which could also indirectly influence immunity, and the remaining87(55%) were involved in a variety of PWBC biological processes that might eventually influence the immune function. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that the DEGs were predominantly enriched in seven immune-related pathways. Hierarchical cluster analysis identified gene expression patterns that were distinguishable between the two study groups.CONCLUSION: Our results suggest that T2DM-SDP patients experience significant hypoimmunity and/or immune dysfunctions, and possess a specific gene expression profile. These findings offer new insights into SDP and the clinical pattern differentiation of T2DM-SDP.

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.

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.