Basic Biological Research On Constitution Of Yin Deficiency And Yang Deficiency Based On Protein Chip Technology

Objiective:By researching、comparing and analyzing the characteristics and differences in proteomics between Yin-deficiency constitution and Yang-deficiency constitution,and discussing the protein expression signaling pathway related with Yin-deficiency constitution and Yang-deficiency constitution,Ultimately providing biological explanation for "Adjustable constitution" theory.Methods:1.All subjects are from the natural population of Beijing,gotten in the Physical examination center and Guo Yi Tang of Beijing University of Chinese Medicine.2.According to the "Chinese Medical Association standards Chinese constitution classification and determination" promulgated and implemented in 2009,screened aged 25 to 40 years of age,regular Physical examination is normal or no clear diagnosis and comply with Normal constitution、Yin-deficiency and Yang-deficiency by criteria select 12 cases of Pinghe constitution,12 cases of Yin-deficiency constitution,12 cases of Yang-deficiency constitution.3.Detect the Fastingsubjects’ serum with AAH-BLG-1000 kit,the use of the instrument comes with software to extract data with AAH-BLG-1000 data analysis software for data analysis.Results:1.By comparing protein microarray results of Yin-deficiency constitution and Normal constitution,we found different proteins of Yin-deficiency and Normal constitution in energy metabolism and neuroendocrine(fold change>5,fluorescent signal>250)are 39 were NF1,TYRO10,IL-27,FGFR2,ApoC1,KLF4,INSRR,GLP-1,Clusterin,Smad 7,SSTR2,MMP-9,CBP,SERPING1,Itk,ApoE3,Flt-3 Ligand,ADAMTS-L2,CCK,Neurokinin-A,FGFR1,RET,EphA4,PI 3Kinase p85 beta,LPS,C-peptide,C5/C5a,FGF R4,IGF-II,CXCR4(fusin),M-CSF,Osteopontin,IL-17F,HB-EGF,RYK,OSM,CART,IL-29,APRIL,which regulate the expression of 27,12 down-regulated expression;we found different proteins of Yang-deficiency and Normal constitution in energy metabolism and neuroendocrine(fold change>5,fluorescent signal>250)there are 39 NF1,INSRR,GLP-1,MCP-1,pro-Glucagon,Alpha 1 Microglobulin,Smad 7,INSL3,SHBG,hCG alpha,Procalcitonin,ADAMTS-1,ADAMTS-L2,VEGF,RET,HE4,Neurokinin-A,GREMLIN,Thyroglobulin,IFN-gamma,GDF8,C-peptide,Growth Hormone(GH),GDF9,corticosteroid-binding globulin,IL-2,ProSAAS,SSTR5,CD40 Ligand/TNFSF5/CD154,Wilms Tumor 1,M-CSF,IGF-II,CXCR4(fusin),OSM,Presenilin 2,uPA,Erythropoietin R,CART,BMPR-IB/ALK-6,which regulate the expression of 18,10 down-regulated expression;yin and yang constitution protein chip are compared and found 77 difference protein,wherein Fold change≥1.5,Fold change<0.7 a total of 18 proteins,and Fold change<0.5 has two.2.By GO functional annotation of the differential proteins,GO function of Yin-deficiency constitution in energy metabolism and neuroendocrine aspects related mainly GO:0045664～regulation of neuron differentiation,GO:0050767～neural regulation,GO:0007218～nerve signals pathways,GO:0030182～neuronal differentiation GO:0048666～neuronal development GO:0031644～nervous system process regulation,GO:0008585～female gonadal development,GO:0010817～regulate hormone levels,GO:0031960～response to corticosteroid,GO:0050707～cytokine secretion regulation,GO:0051384～response to glucocorticoid,GO:0046888～negative regulation of hormone secretion,GO:0032570～progesterone stimulation,GO:0008406～gonadal development,GO:0001655～genitourinary system development,GO:0046883～regulate hormone secretion,GO:0043627～estrogen stimulation,GO:0051046～secretion regulation;GO:metabolic process 0008202～steroids,GO:0045936～negative regulation of phosphorus metabolism,GO:0019217～regulation of fatty acid metabolism,GO:0006140～nucleotide metabolism regulation,GO:0030799～cyclic nucleotide metabolism regulation,GO:0006641一 triglycerides ester metabolism,GO:0045935～bases,nucleosides,nucleotides and nucleic acid metabolism in the process of positive regulation,GO:0019216～regulation of lipid metabolism,GO:0010604～macromolecules positive regulation of metabolic processes,GO:0051247～protein metabolism during positive regulation,GO:0032268～cellular protein metabolism regulation,GO:0051174～phosphorus metabolism regulation,GO:0019220～phosphate metabolism regulation and control;and Yang and physical energy metabolism and nerve GO endocrine function associated mainly GO:0045664～neuronal differentiation regulation,GO:0050767～neural regulation,GO:0007218～nerve signal transduction pathways,GO:0031646～neurological process positive regulation,GO:0030182～neurons differentiation,GO:0048666～development of neurons,GO:process 0031644～nervous system regulation,GO:0008585～female gonadal development,GO:0010817～regulate hormone levels,GO:0031960～corticosteroids irritation,GO:0050707～regulation of cytokine secretion,GO:0051384～glucocorticoids stimulate answer,GO:0046888～negative regulation of hormone secretion,GO:0032570～progesterone stimulation,GO:0008406～gonadal development,GO:0051048～negative regulation of secretion GO:0046883～regulate hormone secretion,GO:0043627～estrogen stimulation response,GO:0050708～protein secretion regulation,GO:0051046～secretion regulation;GO:regulation of metabolic processes 0019218～steroids,GO:0010562～phosphorus positive regulation of metabolic processes,GO:0010563～phosphorus metabolism of negative regulation,GO:0010605～macromolecule metabolic process of negative regulation,GO:0019217～regulation of fatty acid metabolism,GO:0006639～neutral fat metabolism,GO:nucleotide metabolism regulation 0006140～,GO:0030799～cyclic nucleotide metabolism regulation,GO:0006641～triglyceride metabolism,GO:0045935～bases,nucleosides,nucleotides,and nucleic acid metabolism in the process of positive regulation,GO:nitrogen compound metabolic processes 0051173～positive regulator,GO:0051247～protein metabolism during positive regulation,GO:0051248～protein metabolism in the process of negative regulation,GO:0006796～phosphate ester metabolism,GO:0010604～macromolecules positive regulation of metabolic processes,GO:0032270～cellular proteins positively regulate metabolic processes;yin and yang constitution differences protein-related GO function of the extracellular space,blood particles,outer membrane,leukocyte migration,receptor binding,external stimuli,injury response,vesicles bounded cytoplasmic membrane vesicles,the film side,secretory granules chamber,regulation of enzyme activity,platelet αgranule chamber,cell migration,endopeptidase activity regulation,regulation of noxious reaction,spherical high-density lipoprotein particles,inflammation,platelet degranulation,extracellular matrix tissue,extracellular tissue,platelet a particles endopeptidase activity of a negative regulator of hydrolysis activity negative control,a negative regulator of cell migration,cell localization,peptidase activity of a negative regulator of lipid transport,chemotaxis,chylomicrons,multi-cellular biological processes are regulated angiogenesis,acute inflammation,protein processing,protein processing regulation,regulation of macrophage-derived foam cell differentiation,extracellular matrix binding,regulate cell adhesion,cytokine response,macrophage-derived foam cells,foam cells,vascular morphology,reverse cholesterol transport;cell-matrix adhesion,very low density lipoprotein particles reshape the regulation of lipid positioning,wound healing,plasma lipoprotein particles,shaped cell adhesion,vascular development,lipid transport regulation.3.By signaling pathways annotation in KEGG of the differential proteins found Yin-deficiency constitution associated with cytokine signaling pathway and its main receptor interaction pathway(Cytokine-cytokine receptor interaction),PI3K-Akt signaling pathway(PI3K-Akt signaling pathway),JAK-STAT signaling pathway(JAK-STAT signaling pathway);Yang-deficiency constitution associated with the main differences in protein signaling pathways and cytokine receptor interaction pathway(cytokine-cytokine receptor interaction),PI3K-Akt signaling pathway(PI3K-Akt signaling pathway),TGF-beta signaling pathway(TGF-beta signaling pathway);yin and yang constitution associated signaling pathways mainly of cytokine receptor interaction pathway(cytokine-cytokine receptor interaction),PI3K-Akt signaling pathway(PI3K-Akt signaling pathway).4.Differences protein for protein interaction network study found that key proteins for Yin-deficiency constitution are mainly INS,EGFR,VEGF,FGFR2,the key proteins for YYang-deficiency constitution are mainly INS,EGFR,VEGF,PIK3R1,KIT.Conclusion:In this study,we do proteomics perspective comparative study of Yin-deficiency constitution、Yang-deficiency constitution and Normal constitution,found there are significant differences in terms of energy metabolism and neuroendocrine between Yin-deficiency constitution、Yang-deficiency constitution and Normal constitution.In addition,the differential protein is related to the function of immune function,cell cycle,inflammation and other functions.The differences protein between Yin-deficiency constitution and Normal constitutionYin constitution and mainly involved in cytokine and cytokine receptor interaction pathway,PI3K-Akt signaling pathway,JAK-STAT signaling pathway.The PI3K-Akt signaling pathway is closely related to metabolic and hypothalamus-pituitary-gonadal axis,mainly reflected in the role of insulin and sex hormone.The JAK-STAT signaling pathway influence yin deficiency constitution through the effects of leptin;Yang constitution and peaceful constitutional differences between the main protein involved in cytokine and cytokine receptors interaction pathways,PI3K-Akt signaling pathway,TGF-beta signaling pathway.The differences protein between Yang-deficiency constitution and Normal constitution mainly involved in cytokine and cytokine receptor interaction pathway,PI3K-Akt signaling pathway,TGF-beta signaling pathway.The first two pathways is the same with yin deficiency constitution,but there are differences between the involvement protein,and TGF beta signaling pathway have very strong immune inhibitory factor,which may affect the low immunity of Yang deficiency constitution,and relate to cold intolerance.Among them,INS,EGFR,VEGF,FGFR2 are the critical nodes of the differences protein interactions between Yin-deficiency constitution and Normal constitution;DLL1,EphB6,PF4/CXCL4,Furin,Integrin alpha V,Galectin-7,Beta IG-H3,CD45,CEACAM-1,TIMP-3,SSTR2,Persephin,PTHLP,PI 3Kinase p85 beta,HSP70,Hck,Activin RIA/ALK-2,Osteopontin.These 18 different proteins can be used to distinguish between Yin-deficiency constitution and Yang-deficiency constitution,If we want to determine the specificity protein of the both constitution,parallel comparative study remains should be done with the other six constitution.