Differential proteomic analysis of the zebrafish neuron protective effect of Cerebralcare granules using iTRAQ labeling coupled with 2DLC-MS/MS / 中国药学杂志

ABSTRACT

OBJECTIVE: To screen the differentially expressed proteins before and after administration of Cerebralcare granules to zebrafish central nerve injury (CNI) models and search practical markers and explore the molecular mechanism of the treatment. METHODS: Isobaric tags for relative and absolute quantitation (iTRAQ) coupled with nano liquid chromatography-tandem mass spectrometry (Nano-LC-MS/MS) were used to analyze and identify differentially expressed serum proteins in the two groups. Bioinformatics was used to analyze the identified differentially expressed proteins, and the expression of representative differential proteins was verified by Western blotting. RESULTS: With the high throughput proteomic technology of iTRAQ coupled with Nano-LC-MS/MS, 1 933 unique proteins were identified, and 130 proteins showed ≥ 1.50 or ≤ 0.70 folds of changes during differentiation. The proteins detected in the zebrafish neuroendocrine brain had roles in the biological processes of translation, metabolic process and neuronal ion channel clustering. Ef1-α (elongation factor 1-alpha) and α-6-F (Na+/K+ transporting ATPase alpha 1 polypeptide) were validated by Western blotting. The two sets of data showed a statistically significant difference (P<0.05). These RESULTS were consistent with those from quantitative mass spectrometry. CONCLUSION: A high-throughput screen for zebrafish central nerve injury proteins can be performed by a combined use of iTRAQ and Nano-LC-MS/MS, and the molecular mechanism of Cerebralcare granules treatment can thus be preliminarily explored.