| Objective:The aim of this study was to screen the differentially expressed proteins in childhood disorders of cortical development (DCD) accompanied epilepsy for exploring potential targets of biomarkers and biologial therapy. Methods:The specimens were childhood disorders of cortical development (DCD) with epilepsy brain tissues (the DCD group) and children normal brain tissues operated for increased intracranial pressure due to head trauma (the control group). The proteins were labeled using relative and absolute quantification (iTRAQ) method, separated and analyzed by 2D liquid chromatograph electrospray tandem mass spectrometry(2D-LC-MS/MS) for identifying the differentially expressed proteins in childhood DCD with epilepsy brain tissues. The differentially expressed proteins were analyzed by bioinformatics for screening the valueable proteins. The screened proteins were further validated using Real-time quantitative Polymerase Chain Reaction (qPCR), immunoblotting and immunohischemical analysis. Results:According to using iTRAQ and 2D-LC-MS/MS method, a total of 153 proteins were different expressed significantly in childhood DCD with epilepsy brain tissues (64 highly expressed proteins and 89 decreased expressed proteins; P<0.05). According to GO (Gene ontology) analysis, we found that these differentially expressed proteins were mainly involved in catalytic activity. The protein interaction network showed transferri (TF), apolipoprotein A1 (APOA1), alpha-2-macroglobulin (A2M), ATPase (ATP6V1A), vimentin (VIM) were located in the connected notes. The epression of TF, APOA1, A2M, ATP6V1A, VIM were further vertified by qPCR, Western blotting and immunohischemical, the results were in keep with the results of quantitative mass spectrometry. Conclusion:The differentially expressed proteins can be effectively screened by iTRAQ combined with 2D-LC-MS/MS, and may provide new clues for potential biological biomarkers and therapeutic targets in childhood DCD with epilepsy. |
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