Amadori-modified glycated albumin predominantly induces E-selectin expression on human umbilical vein endothelial cells through NADPH oxidase activation
Amadori-modified glycated albumin predominantly induces E-selectin expression on human umbilical vein endothelial cells through NADPH oxidase activation
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DOI：
10.1016/j.cca.2005.12.008
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发表时间：
2006-05-01
影响因子：
5
通讯作者：
Matsumoto, K
中科院分区：
文献类型：
--
作者：
Higai, K;Shimamura, A;Matsumoto, K
背景：蛋白糖化与糖尿病患者的内皮细胞功能障碍和血管并发症密切相关。糖化白蛋白诱导的细胞信号转导类似于晚期糖基化内皮细胞(AGEs)，但细胞信号转导机制尚不清楚。方法：用糖化人血清白蛋白(GLC-HSA)刺激人脐静脉内皮细胞(HUVECs)，用实时定量聚合酶链式反应(RT-PCR)和免疫计量学方法检测E-选择素的表达，用多种信号分子抑制剂和共聚焦显微镜检测细胞信号转导。结果：GLC-HSA诱导的E-选择素表达是3种糖基化内皮细胞(AGEs-HSA)的10~20倍，而抗AGEs受体(RAGE)抗体不能抑制其表达。NADPH氧化酶抑制剂二苯基碘氯和活性氧清除剂N-乙酰-L半胱氨酸可完全抑制Glc-HSA诱导的E-选择素表达。共聚焦显微镜分析也显示ROS在细胞内积聚。磷脂酰肌醇3激酶(PI3K)抑制剂Wortmannin和LY294002、蛋白激酶B(PKB)抑制剂ML-9、I kappaB(IKK)抑制剂Bay 117082和Jun N末端激酶(JNK)抑制剂SP600125均可抑制Glc-HSA诱导的E-选择素表达，而蛋白激酶C(Calphostin C)和H-7不抑制Glc-HSA诱导的E-选择素表达。结论：Glc-HSA可诱导NADPH氧化酶、PKB-IKK和JAP-1的激活，进而上调E-选择素基因转录。(C)2005爱思唯尔B.V.保留所有权利。
Background: Protein glycation is closely linked to endothelial-cell dysfunction and vascular complications in diabetes. Glycated albumin is reported to induce cellular signaling similar to advanced glycation endoproducts (AGEs), however, cellular signaling remains obscure.Method: We stimulated human umbilical vein endothelial cells (HUVECs) by glycated human serum albumin (Glc-HSA), determined E-selectin expression by real-time PCR and immunometric methods, and estimated cellular signaling by using various signaling molecule inhibitors and confocal microscopy.Results: Glc-HSA-induced E-selectin expression was 10 or 20 times more than that induced with 3 kinds of AGEs-HSAs, which was not suppressed by anti-receptor for AGEs (RAGE) antibody. Glc-HSA-induced E-selectin expression was completely suppressed by the NADPH oxidase inhibitor diphenylene iodonium chloride and the reactive oxygen species (ROS) scavenger N-acetyl-L-cysteine. Confocal microscopic analysis also revealed intracellular accumulation of ROS. Glc-HSA-induced E-selectin expression was suppressed by the phosphatidylinositol 3 kinase (PI3K) inhibitors wortmannin and LY294002, the protein kinase B (PKB) inhibitor ML-9, the I kappa B kinase (IKK) inhibitor BAY 117082, and the Jun N-terminal kinase (JNK) inhibitor SP600125, On the other hand, the protein kinase C inhibitors calphostin C and H-7 did not suppress Glc-HSA-induced E-selectin expression.Conclusion: Glc-HSA induces activation of NADPH oxidase, PKB-IKK and JNK, then E-selectin gene transcription is upregulated by nuclear-translocated NF-kappa B and AP-1. (c) 2005 Elsevier B.V All rights reserved.