Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune 411008, India; Academy of Scientific and Innovative Research (AcSIR), India.
Proteomics Facility, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Pune 411008, India.
J Proteomics. 2018 Sep 15;187:69-79. doi: 10.1016/j.jprot.2018.06.009. Epub 2018 Jun 21.
Endothelial dysfunction is one of the primary steps in the development of diabetes associated cardiovascular diseases. Hyperglycemic condition in diabetes promotes accumulation of advanced glycation end products (AGEs) in the plasma, that interact with the receptor for AGEs (RAGE) present on the endothelial cells and negatively affect their function. Using Human umbilical vascular endothelial cells (HUVECs) in culture, the effect of glycated human serum albumin on global proteomic changes was studied by SWATH-MS, a label free quantitative proteomic approach. Out of the 1860 proteins identified, 161 showed higher abundance while 123 showed lesser abundance in cells treated with glycated HSA. Bioinformatic analysis revealed that the differentially regulated proteins were involved in various processes such as apoptosis, oxidative stress etc. that are associated with endothelial dysfunction. Furthermore, the iRegulon analysis and immunofuorescence studies indicated that several of the differentially regulated proteins were transcriptionally regulated by NF-κB, that is downstream to AGE-RAGE axis. Some of the important differentially regulated proteins include ICAM1, vWF, PAI-1 that affect important endothelial functions like cell adhesion and blood coagulation. qPCR analysis showed an increase in expression of the AGE receptor RAGE along with other genes involved in endothelial function. AGE treatment to HUVEC cells led to increased oxidative stress and apoptosis. This is the first proteomics study that provides insight into proteomic changes downstream to AGE-RAGE axis leading to endothelial dysfunction and predisposing to cardiovascular complications.
Cardiovascular disease (CVD) is a major pathological outcome in diabetic patients and it is important to address ways that target its development before the onset. Elevated plasma AGEs in diabetes can affect endothelial function and can continue to show their effects even after blood glucose levels are back to normal. Since endothelial dysfunction acts as one of the initiating factors for the development of CVD, understanding how AGEs affect the endothelial cell proteome to cause dysfunction will provide insight into the mechanisms involved and aid designing new therapeutic approaches.
探讨糖化白蛋白对人脐静脉内皮细胞（HUVEC）整体蛋白质组变化的影响。
采用 SWATH-MS 无标记定量蛋白质组学方法，研究了糖化人血清白蛋白对培养的 HUVEC 中蛋白质组变化的影响。
在鉴定出的 1860 种蛋白质中，有 161 种在糖化 HSA 处理的细胞中丰度较高，而 123 种丰度较低。生物信息学分析表明，差异调节蛋白参与了与内皮功能障碍相关的各种过程，如细胞凋亡、氧化应激等。此外，iRegulon 分析和免疫荧光研究表明，几种差异调节蛋白受 NF-κB 的转录调控，即 AGE-RAGE 轴的下游。一些重要的差异调节蛋白包括影响细胞黏附和血液凝固等重要内皮功能的 ICAM1、vWF 和 PAI-1。qPCR 分析显示 AGE 受体 RAGE 及其它参与内皮功能的基因表达增加。AGE 处理 HUVEC 细胞导致氧化应激和细胞凋亡增加。这是第一项关于 AGE-RAGE 轴下游导致内皮功能障碍和易患心血管并发症的蛋白质组学研究。
心血管疾病（CVD）是糖尿病患者的主要病理后果，在发病前确定针对其发展的方法非常重要。糖尿病患者血浆 AGEs 升高会影响内皮功能，即使血糖水平恢复正常，其影响仍会持续存在。由于内皮功能障碍是 CVD 发展的起始因素之一，了解 AGE 如何影响内皮细胞蛋白质组以导致功能障碍，将为相关机制提供深入了解，并有助于设计新的治疗方法。
