Vaspin inhibited proinflammatory cytokine induced activation of nuclear factor-kappa B and its downstream molecules in human endothelial EA.hy926 cells
Vaspin inhibited proinflammatory cytokine induced activation of nuclear factor-kappa B and its downstream molecules in human endothelial EA.hy926 cells
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Vaspin 抑制人内皮 EA.hy926 细胞中促炎细胞因子诱导的核因子 kappa B 及其下游分子的激活
DOI：
10.1016/j.diabres.2013.12.002
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发表时间：
2014-03-01
影响因子：
5.1
通讯作者：
Zheng, Caihong
中科院分区：
文献类型：
--
作者：
Liu, Shiwei;Dong, Yanting;Zheng, Caihong
目的：在这项研究中，我们研究了内脏脂肪组织来源的丝氨酸蛋白酶抑制剂（vaspin），一种新发现的脂肪细胞因子，对核因子-κ B的影响（NF-κ B）及其下游分子在促炎细胞因子、肿瘤坏死因子-α（TNF-α）和白细胞介素-1（IL-1）刺激人内皮EA.hy926细胞，以阐明vaspin在内皮炎症状态中的作用。构建NF-κ B荧光素酶报告系统并稳定转染人内皮细胞株EA.hy926。转染后，在TNF-α和IL-1刺激前，用各种浓度的vaspin（0-320 ng/ml）预处理EA.hy926细胞。用荧光素酶报告基因检测NF-κ B的转录活性。通过酶联免疫吸附测定（ELISA）测量NF-κ B B下游炎性细胞因子TNF-α、IL-1和IL-6的表达水平。采用实时荧光定量PCR（RT-PCR）和蛋白质印迹法分别检测细胞间粘附分子-1（ICAM-1）、血管细胞粘附分子-1（VCAM-1）和单核细胞趋化蛋白-1（MCP-1）在mRNA和蛋白质水平的表达。结果显示vaspin浓度依赖性地抑制TNF-α和IL-1介导的NF-κ B及其下游分子的活化（P < 0.05）。结论：我们的结论是vaspin通过抑制NF-κ B及其下游分子保护内皮细胞免受促炎细胞因子诱导的炎症。(C)2013爱思唯尔爱尔兰有限公司版权所有。
Aims: In this study, we investigated the effects of visceral adipose tissue-derived serpin (vaspin), a newly discovered adipocytokine, on nuclear factor-kappa B (NF-kappa B) and its downstream molecules in proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha) and interleukine-1 (IL-1), stimulated human endothelial EA.hy926 cells to elucidate the role of vaspin in the inflammatory states of endothelium.Methods: A NF-kappa B luciferase reporter system was constructed and stably transfected into human endothelial cell line EA.hy926. Following transfection, EA.hy926 cells were pre-treated with various concentrations of vaspin (0-320 ng/ml) before TNF-alpha and IL-1 stimulation. The transcription activity of NF-kappa B was determined using luciferase reporter assay. Expression levels of NF-kappa B downstream inflammatory cytokines, TNF-alpha, IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Expressions of adhesion molecules and chemokines, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemotactic protein-1 (MCP-1) were determined by quantitative real-time PCR (RT-PCR) and western blot in mRNA and protein levels, respectively.Results: Results showed that vaspin inhibited TNF-alpha and IL-1 mediated activation of NF-kappa B and its downstream molecules in a concentration-dependent manner (P < 0.05). Conclusions: We conclude that vaspin protected endothelial cells from proinflammatory cytokines induced inflammation by inhibition of NF-kappa B and its downstream molecules. (C) 2013 Elsevier Ireland Ltd. All rights reserved.