Study on the Application of iTRAQ Technology in Retinal Apoptosis Protein in Diabetic Retinopathy Mice
QUE Li-dan1, LIN Yi-long1, QIU Xiu-ping1, LIU Bin2
1. Longyan First Affiliated Hospital of Fujian Medical University, Longyan Fujian 364000, China; 2. Quanzhou Maternal and Child Health Hospital Children's Hospital, Quanzhou Fujian 362000, China
Study on the Application of iTRAQ Technology in Retinal Apoptosis Protein in Diabetic Retinopathy Mice
QUE Li-dan1, LIN Yi-long1, QIU Xiu-ping1, LIU Bin2
1. Longyan First Affiliated Hospital of Fujian Medical University, Longyan Fujian 364000, China; 2. Quanzhou Maternal and Child Health Hospital Children's Hospital, Quanzhou Fujian 362000, China
摘要Objective: Using isobaric tags for relative and absolute quantitation(iTRAQ) technology to study differential protein expression in the retinal tissue of DR mouse models, providing proteomic evidence at the protein level for the pathogenesis of DR. Methods: Firstly, establish diabetic mice and DR mice models, use iTRAQ technology to detect the retinal tissue samples of normal control group and DR model group mice, label the total retinal proteins of mice with iTRAQ reagent, and analyze them using mass spectrometry technology. Evaluate the differential proteins of the two groups using BioWorks TM 3.0 software, and conduct interaction feature analysis on the different proteins using the STRING website. Results: The iTRAQ technology detected a total of 406 different proteins between the diabetes and normal control groups, with 19 of them closely related to retinal cell apoptosis. Among them, significantly different proteins include acyl-CoA dehydrogenase short chain specific(ACADS), ataxin-10(ATXN10), BCL-2-associated X protein(BAX), caspase-3(CASP3), collagen type II α1 chain(COL4A2), glycyl-tRNA synthetase(GARS), glial fibrillary acidic protein(GFAP), legumain(LGMN), mucin-4(MUC4), N-myc downstream-regulated gene 1 protein (NDRG1), with ratios to internal controls in the normal group of 1.67, 2.06, 1.76, 2.16, 1.53, 1.87, 1.24, 1.61, 0.42, 0.56, respectively; and there is a potential functional association between GFAP, CASP3, and BAX proteins. Conclusion: In DR mice retinas, there are abnormal changes in the expression of a large number of apoptosis-related proteins. iTRAQ technology can effectively screen out key apoptosis proteins, among which GFAP, CASP3, and BAX may have adverse effects on the progression of DR by participating in the apoptosis process. The application of iTRAQ technology can provide new technical support for proteomic research on apoptosis proteins.
Abstract：Objective: Using isobaric tags for relative and absolute quantitation(iTRAQ) technology to study differential protein expression in the retinal tissue of DR mouse models, providing proteomic evidence at the protein level for the pathogenesis of DR. Methods: Firstly, establish diabetic mice and DR mice models, use iTRAQ technology to detect the retinal tissue samples of normal control group and DR model group mice, label the total retinal proteins of mice with iTRAQ reagent, and analyze them using mass spectrometry technology. Evaluate the differential proteins of the two groups using BioWorks TM 3.0 software, and conduct interaction feature analysis on the different proteins using the STRING website. Results: The iTRAQ technology detected a total of 406 different proteins between the diabetes and normal control groups, with 19 of them closely related to retinal cell apoptosis. Among them, significantly different proteins include acyl-CoA dehydrogenase short chain specific(ACADS), ataxin-10(ATXN10), BCL-2-associated X protein(BAX), caspase-3(CASP3), collagen type II α1 chain(COL4A2), glycyl-tRNA synthetase(GARS), glial fibrillary acidic protein(GFAP), legumain(LGMN), mucin-4(MUC4), N-myc downstream-regulated gene 1 protein (NDRG1), with ratios to internal controls in the normal group of 1.67, 2.06, 1.76, 2.16, 1.53, 1.87, 1.24, 1.61, 0.42, 0.56, respectively; and there is a potential functional association between GFAP, CASP3, and BAX proteins. Conclusion: In DR mice retinas, there are abnormal changes in the expression of a large number of apoptosis-related proteins. iTRAQ technology can effectively screen out key apoptosis proteins, among which GFAP, CASP3, and BAX may have adverse effects on the progression of DR by participating in the apoptosis process. The application of iTRAQ technology can provide new technical support for proteomic research on apoptosis proteins.
QUE Li-dan, LIN Yi-long, QIU Xiu-ping, LIU Bin. Study on the Application of iTRAQ Technology in Retinal Apoptosis Protein in Diabetic Retinopathy Mice[J]. 中国生物医学工程学报(英文版）, 2023, 32(4): 161-169.
QUE Li-dan, LIN Yi-long, QIU Xiu-ping, LIU Bin. Study on the Application of iTRAQ Technology in Retinal Apoptosis Protein in Diabetic Retinopathy Mice. Chinese Journal of Biomedical Engineering, 2023, 32(4): 161-169.
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