Comparative Proteomics Analysis Of Cerebrospinal Fluid Of Patients With Guillain-Barre' Syndrome

Guillain-Barre syndrome(GBS)is an immunemediated inflammatory disease of the peripheral nerves involving both the myelin sheath and the axons.GBS occurs worldwide especially in developing country and with an often severe course and poor prognosis.Even when patients are treated in well-equipped intensive care units, mortality rates are still 3-7%.Moreover,about 7-15%of patients have permanent neurological sequelae,for example bilateral footdrop,intrinsic hand muscle wasting, sensory ataxia and dysesthesia,which severely affect the patients daily life. However,the molecular mEechanisms underlying the disease remain poorly understood and so far no reliable disease-related markers are available.Lots of studies suggesed there are different expression in CSF of patients with GBS. Increased protein level without pleocytosis in the CSF is a characteristic of patients with GBS,which suggest that Cerebrospinal fluid(CSF)is a promising source of proteins that may provide important information about the pathomechanisms in GBS.Now,the proteomic approach for identification and quantification of the entire protein content(proteome)of the organism is helpful in discovery of biomarkers. Therefore,the study of proteome of CSF will provide new insight into the pathomechanisms underlying GBS.In this study,By mean of proteomic analysis with high-resolution two-dimensional gel electrophoresis(2-DE)followed by mass spectrometry(MS)and database searching,the proteins which may be specifically up or down regulated in patients with GBS maybe provided as disease-related proteins to both aid in the diagnosis of GBS and possibly its treatment and prognosis.The subsequent Western-blot and ELISA to confirme the result of proteomics.Objective:To establish comparative proteomics technology system for cerebrospinal fluid(CSF)of patients with Guillain-Barre'syndrome(GBS),and to find differences of CSF proteins between healthy people and patients with GBS.These identified proteins may be involved in the pathophysiological process of GBS and call for further studying the role of these proteins in the pathogenesis of the disease.Methods:CSF were extracted from patients with GBS and controls,and the proteins were precipitated with ice-cold acetone.Isoelectric focusing electrophoresis(IEF)was employed as the first dimension and sodium dodecylsulfate—polyacrylamide gel electrophoresis(SDS-PAGE)was employed as the second dimension.The two-dimensional analysis gels maps were analysed by Imagemaster 2D software to find obvioudly alterated proteins,and attained the proteins in the prepare gels.The peptides mass fingerprint was got by MOLDI/TOF/MS.To search in SWISSPROT database to identify differential proteins. And the interested proteins to be confirmed by subsequent ELISA.Results:2-dimensional maps of CSF with GBS and the controls group were got. 387±29(experiment group)and 398±36(control group)protein spots were identified. There are obvious difference between the two group,And 10 protein spots had significant difference in expression.Expression of 4 protein spots in GBS group increased and 6 decreased.Cystatin C which is the most significantly decrease protein was discoveried firstly in this study,The subsequent ELISA also confirmed the concentration of Cystatin C was significantly lower than that of control group.Conclusion:Proteomics technology system including 2DE and Bio-Mass Spectrometry was established successfully to analysis CSF with GBS.6 special proteins relevant to GBS were identified and the lower expression of Cystatin C in CSF of patients with GBS was Validated by immunological methods.Theses can offers clues for study on machanisms of GBS.