PDF(Pubmed)
Abstract：
BACKGROUND: The purpose of this study is to identify the proteomic differences between the aqueous humour of diabetes patients with cataracts and that of non-diabetic sufferers of cataracts in a clinical setting.
METHODS: Patients were divided into the diabetic experimental group and the non-diabetic control group. Aqueous humour specimens were obtained via cataract surgery. Sample proteins were treated with a TMT reagent, separated using a cation chromatography column, and analysed using a C18 desalting column. Proteins were identified using HPLC-MS/MS. The differential proteins were identified using both a p value of < 0.05 and a fold change of > 1.2. GO classification enrichment analysis, KEGG pathway enrichment analysis, protein interaction network analysis, and ingenuity pathway analysis were all carried out. The expression level of four differential proteins were verified by Western blot, and GC and TTR expressions were further examined using an expanded sample pool.
RESULTS: The postprandial glucose levels between the experimental group (9.40 ± 1.35 mmol/L) and the control group (6.56 ± 0.81 mmol/L) were significantly different, with a p value of 1.16E-06. It is important to note, however, that the baseline levels of the parameters showed no statistical differences. In total, 397 aqueous humour proteins were identified; of these, 137 showed significant differences, with 63 upregulated ones and 74 down-regulated ones. The differential proteins play important roles in numerous biological processes and pathways, such as complement and coagulation cascades (p = 1.71E-09). Some of these differential proteins are associated with diabetic retinal degeneration and other diabetic complications. Differential proteins, such as HP, GC, and TTR, have high node degree in the protein interaction network. Western blot results further confirmed that GC were down-regulated while TTR was up-regulated in aqueous humour under diabetic condition.
CONCLUSIONS: A list of differential proteins in the human aqueous humour of diabetic patients was established. Proteins with high interaction scores as per protein interaction analysis, such as GC and TTR, were further verified and could potentially be used as early diagnostic markers for diabetic eye complications in clinical practice.
METHODS: Patients were divided into the diabetic experimental group and the non-diabetic control group. Aqueous humour specimens were obtained via cataract surgery. Sample proteins were treated with a TMT reagent, separated using a cation chromatography column, and analysed using a C18 desalting column. Proteins were identified using HPLC-MS/MS. The differential proteins were identified using both a p value of < 0.05 and a fold change of > 1.2. GO classification enrichment analysis, KEGG pathway enrichment analysis, protein interaction network analysis, and ingenuity pathway analysis were all carried out. The expression level of four differential proteins were verified by Western blot, and GC and TTR expressions were further examined using an expanded sample pool.
RESULTS: The postprandial glucose levels between the experimental group (9.40 ± 1.35 mmol/L) and the control group (6.56 ± 0.81 mmol/L) were significantly different, with a p value of 1.16E-06. It is important to note, however, that the baseline levels of the parameters showed no statistical differences. In total, 397 aqueous humour proteins were identified; of these, 137 showed significant differences, with 63 upregulated ones and 74 down-regulated ones. The differential proteins play important roles in numerous biological processes and pathways, such as complement and coagulation cascades (p = 1.71E-09). Some of these differential proteins are associated with diabetic retinal degeneration and other diabetic complications. Differential proteins, such as HP, GC, and TTR, have high node degree in the protein interaction network. Western blot results further confirmed that GC were down-regulated while TTR was up-regulated in aqueous humour under diabetic condition.
CONCLUSIONS: A list of differential proteins in the human aqueous humour of diabetic patients was established. Proteins with high interaction scores as per protein interaction analysis, such as GC and TTR, were further verified and could potentially be used as early diagnostic markers for diabetic eye complications in clinical practice.
摘要：
背景：本研究的目的是确定糖尿病白内障患者和非糖尿病白内障患者的房水之间的蛋白质组学差异。
方法：患者分为糖尿病实验组和非糖尿病对照组。房水标本是通过白内障手术获得的。样品蛋白用TMT试剂处理,使用阳离子色谱柱分离，并使用C18脱盐柱进行分析。使用HPLC-MS/MS鉴定蛋白质。使用<0.05的p值和>1.2的倍数变化鉴定差异蛋白。GO分类富集分析,KEGG途径富集分析,蛋白质相互作用网络分析，并进行了信号通路分析。通过Westernblot验证四种差异蛋白的表达水平，使用扩展的样品池进一步检查GC和TTR表达。
结果:实验组(9.40±1.35mmol/L)与对照组(6.56±0.81mmol/L)餐后血糖水平差异显著,p值为1.16E-06。重要的是要注意,然而,参数的基线水平没有统计学差异。总的来说,鉴定出397种水性体液蛋白；其中，137显示出显著差异，63个上调的和74个下调的。差异蛋白在许多生物过程和途径中起着重要作用,如补体和凝血级联(p=1.71E-09)。这些差异蛋白中的一些与糖尿病视网膜变性和其他糖尿病并发症有关。差异蛋白质，如HP,GC,和TTR,在蛋白质相互作用网络中具有较高的节点度。Westernblot结果进一步证实，在糖尿病状态下，房水中GC下调，而TTR上调。
结论：建立了糖尿病患者的人房水差异蛋白列表。根据蛋白质相互作用分析，具有高相互作用分数的蛋白质，如GC和TTR，进一步验证，并有可能在临床实践中用作糖尿病眼并发症的早期诊断标志物。
方法：患者分为糖尿病实验组和非糖尿病对照组。房水标本是通过白内障手术获得的。样品蛋白用TMT试剂处理,使用阳离子色谱柱分离，并使用C18脱盐柱进行分析。使用HPLC-MS/MS鉴定蛋白质。使用<0.05的p值和>1.2的倍数变化鉴定差异蛋白。GO分类富集分析,KEGG途径富集分析,蛋白质相互作用网络分析，并进行了信号通路分析。通过Westernblot验证四种差异蛋白的表达水平，使用扩展的样品池进一步检查GC和TTR表达。
结果:实验组(9.40±1.35mmol/L)与对照组(6.56±0.81mmol/L)餐后血糖水平差异显著,p值为1.16E-06。重要的是要注意,然而,参数的基线水平没有统计学差异。总的来说,鉴定出397种水性体液蛋白；其中，137显示出显著差异，63个上调的和74个下调的。差异蛋白在许多生物过程和途径中起着重要作用,如补体和凝血级联(p=1.71E-09)。这些差异蛋白中的一些与糖尿病视网膜变性和其他糖尿病并发症有关。差异蛋白质，如HP,GC,和TTR,在蛋白质相互作用网络中具有较高的节点度。Westernblot结果进一步证实，在糖尿病状态下，房水中GC下调，而TTR上调。
结论：建立了糖尿病患者的人房水差异蛋白列表。根据蛋白质相互作用分析，具有高相互作用分数的蛋白质，如GC和TTR，进一步验证，并有可能在临床实践中用作糖尿病眼并发症的早期诊断标志物。
