分类: 医学、药学 >> 基础医学 提交时间: 2017-12-27 合作期刊: 《南方医科大学学报》
摘要: Objective Intravitreal (IVT) injection has become one of the most commonly performed ophthalmologic procedures. We investigated the changes in retinal function and proteomics in rabbits receiving IVT injection of PBS to evaluate the safety of IVT injection. Methods Twenty Chinchilla rabbits were subjected to IVT injection of 50 μL PBS in the right eyes. On days 0, 4, 7 and day 14, the retinas of the rabbits were isolated after routine ophthalmic and electroretinogram examinations. The protein expressions in the retinas were quantified using tandem mass tag (TMT)-labeling coupled with LC-MS/MS, and bioinformatic analysis of the differentially expressed proteins (DEPs) was performed based on KEGG database to identify significantly enriched pathways. Functional network of the significant DEPs was analyzed using STRING. Results No noticeable fundus or functional changes occurred in the rabbit eyes following IVT injection of PBS. A total of 6042 retinal proteins were identified in the retina following the injection, among which 49 proteins (0.81%) exhibited over 5.0-fold up-regulation or over 80% downregulation relative to the control. Most of the distinctly up-regulated or down-regulated proteins were associated with the cytoskeleton. Significantly enriched pathways involved focal adhesion, tight junction, riboflavin metabolism, extracellular matrix-receptor interaction and regulation of actin cytoskeleton. Functional network analysis showed that ACTC1 and ISG15 played central roles in the protein interaction networks. Conclusion IVT PBS injection in rabbits causes alterations in proteins associated with cell adhesion, morphology, migration, differentiation, signal transduction and riboflavin metabolism, but the alterations of the retinal proteins appear not sufficient to cause observable pathology of the retina.
分类: 生物学 >> 植物学 >> 植物生物化学、植物生物物理学 提交时间: 2016-05-04
摘要: Molecular changes elicited by plants in response to fungal attack and how this affects plantpathogen interaction, including susceptibility or resistance, remain elusive. We studied the dynamics in root metabolism during compatible and incompatible interactions between chickpea and Fusarium oxysporum f. sp. ciceri (Foc), using quantitative label-free proteomics and NMR-based metabolomics. Results demonstrated differential expression of proteins and metabolites upon Foc inoculations in the resistant plants compared with the susceptible ones. Additionally, expression analysis of candidate genes supported the proteomic and metabolic variations in the chickpea roots upon Foc inoculation. In particular, we found that the resistant plants revealed significant increase in the carbon and nitrogen metabolism; generation of reactive oxygen species (ROS), lignification and phytoalexins. The levels of some of the pathogenesis-related proteins were significantly higher upon Foc inoculation in the resistant plant. Interestingly, results also exhibited the crucial role of altered Yang cycle, which contributed in different methylation reactions and unfolded protein response in the chickpea roots against Foc. Overall, the observed modulations in the metabolic flux as outcome of several orchestrated molecular events are determinant of plant's role in chickpeaFoc interactions.
分类: 生物学 >> 生物物理学 >> 生物物理、生物化学与分子生物学 提交时间: 2016-05-12
摘要: The integrins, a family of transmembrane proteins, function in cell-to-cell and cell-to-extracellular matrix (ECM) adhesive interactions, and influence cell signaling of cell growth and differentiation. Expression of integrin 6 in three bladder cancer cell lines, HCV29, KK47 and YST1 were quantitatively analyzed by LC-MS using stable isotope labeling by amino acids in cell culture (SILAC), a simple and powerful proteomic strategy. The results showed that the non-invasive bladder cancer cell line KK47 expressed the highest level of integrin alpha 6. The expression of integrin alpha 6 in invasive bladder cancer cell line YTS1 was also higher than in normal bladder epithelial cell line HCV29. Furthermore, these results were confirmed by Western blotting, qPCR, immunohistochemistry and flow cytometry. Clinical data of mRNA 1TGA6 expression pattern from open-access database (www.oncomine.org) showed the same result during bladder cancer progression. All these indicated that integrin alpha 6 is associated with the invasion progress of the bladder cancer. The preliminary data in this study may sparkle the fundamental role of integrin 6 in the research of bladder cancer.
分类: 生物学 >> 生物物理学 提交时间: 2016-05-12
摘要: SILAC is based on direct addition of selected stable isotope amino acids into the cell culture medium, allowing superior quantitative analysis of the cellular proteome compared to other labeling methods. The great advantages of SILAC lie in its straight-forward implementation, quantitative accuracy, and reproducibility over chemical labeling or label-free quantification strategies, favoring its adoption for proteomic research. SILAC has been widely applied to characterize the proteomic changes between different biological samples, to investigate dynamic changes of protein PTMs, to distinguish specific interacting proteins in interaction proteomic analysis, and to analyze protein turnover in the proteome-wide scale. The present review summarizes the principles of SILAC technology, its applications in biological research, and the present state of this technology.