Study Of Peripheral Blood Mononuclear Cells On "GanYang Hua Feng Syndrome" Of Cervical Spondylosis Of Vertebral Artery By Proteomic Method

SUBJECT Using the technology of two-dimensional polyacrylamide gel electrophoresis(2-DE),to establish high resolution 2-DIMENSIONAL eletrophoregrams of "Gan Yang Hua Feng Syndrome(GYHF)"and "Gan Yang Shang Kang Syndrome(GYSK)"of Cervical Spondylosis of Vertebral Artery and the healthy human group, using proteomics to investigate the essence of GYHF Syndrome of Cervical Spondylosis of Vertebral Artery.METHODS Design 3 groups,GYHF of Cervical Spondylosis of Vertebral Artery,GYSK of Cervical Spondylosis of Vertebral Artery and the health human group.Extract total protein from peripheral blood mononuclear cells,and two dimensional electrophoresis was used to separate total proteins.Matrix-Assisted Laser Desorption/ Ionization-Time Of Flight Mass Spectrometry(MALDI-TOF-MS)and searching in SWISS-PORT and MSDB database were used to identify proteins.Validate the CapZ-βof the identified proteins by RT-PCR.RESULTS1.Establishment of 2-DE maps of each group and identification of the diffderent proteins:we obtained the 3 groups' 2-DE maps,after analysis by PDQuest7.3.1 software,we get 587 protein spots in the healthy human group,622 protein spots in the GYHF group,587 protein spots in the GYSK group.Compare with each other,we get 19 different protein spots including 14 kinds proteins.Compare with the healthy human group,there are 16 differential expressing proteins were found in the GYHF electrophoregram,3 up-regulation,10 down-regulation,3 absence.And 12 different expressing proteins in GYSK group,2 up-regulation,6 down-regulation,4 absence.Compare with the GYHF group,there are 16 different protein spots in GYSK group,and 3 identical expressing protein spots in the two groups.The function of identified proteins refer to Apoptosis-correlate protein,antioxidants, signal transduction,cytoskeletal proteins,hypothetical protein and unknown protein.2.Authenticate the expression of CapZ-βby RT-PCR:Compare with the health adult group,the expression of CapZ-βup-regulated in GYSK and GYHF groups(P＜0.05),compare with the GYSK group,the expression of CapZ-βup-regulated in GYHF group(P＜0.05),the same with the result of MS.CONCLUSION1.The expression maps of proteomics in peripheral blood mononuclear cells(PBMCs)of the GYHF and GYSK of Cervical Spondylosis of Vertebral Artery were rudiment established with the technology of 2-DE.2.There are identical expressing proteins of the GYHF and GYSK of Cervical Spondylosis of Vertebral Artery,which indicates that the two Syndromes of the same disease have the same matter substructure. But they are two different syndromes,have the different protein expression maps which indicates that they have different essential connotation.3.The result of RT-PCR same to the result of Proteromics,testified the technology of Proteromics is stabilized and the result is dependablity. CapZ-βboth expressing in the GYHF and GYSK groups,prompt that the GYHF syndrome developed from GYSK syndrome,they have the same matter substructure.But CapZ-βup-regulation expressing in GYHF group compare to the GYSK group,which indicates that the two Syndromes of the same disease(one disease,different stages)have the different essential connotation.The investigation provid the science evidence for the substance of the GYHF syndrome.