| 引用本文: | 吴亚军,苏洁,黄浦俊,陈国杨,陈素红,吕圭源.蒙花苷对TNF-α诱导的血管内皮细胞炎症损伤及TLR4/IκBα/NF-κB信号通路的影响[J].中国现代应用药学,2017,34(5):637-643. |
| WU Yajun,SU Jie,HUANG Pujun,CHEN Guoyang,CHEN Suhong,LYU Guiyuan.Buddleoside Prevents TNF-α-induced Human Aortic Endothelial Cells Inflammatory Injury Through Inhibiting TLR4/IκBα/NF-κB Signaling Pathway[J].Chin J Mod Appl Pharm(中国现代应用药学),2017,34(5):637-643. |
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| 蒙花苷对TNF-α诱导的血管内皮细胞炎症损伤及TLR4/IκBα/NF-κB信号通路的影响 |
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吴亚军1, 苏洁1, 黄浦俊2, 陈国杨2, 陈素红1,2,3, 吕圭源1
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1.浙江中医药大学, 杭州 310053;2.温州医科大学, 温州 325035;3.浙江工业大学, 杭州 310014
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| 摘要: |
| 目的 探讨蒙花苷对血管内皮细胞的保护作用和机制。方法 MTT法检测肿瘤坏死因子（TNF-α）和蒙花苷对人脐静脉内皮细胞（EA.hy926）活性的影响；采用TNF-α（50 ng·mL-1）刺激EA.hy926，加入不同浓度（5，15和25 mmol·L-1）蒙花苷处理24 h，DAPI染色法测定EA.hy926细胞与单核细胞（THP-1）的黏附能力；Western blot法检测细胞中Toll样受体（TLR4）、核因子-kB抑制蛋白（IκBα）、核因子-kB（NF-κB）、细胞间黏附分子-1（ICAM-1）和血管细胞黏附分子-1（VCAM-1）蛋白的表达；RT-PCR法检测ICAM-1、VCAM-1 mRNA的表达；ELISA法检测细胞上清液中白介素-1（IL-1）、白介素-6（IL-6）和白介素-8（IL-8）的含量。结果 蒙花苷可显著抑制TNF-α刺激EA.hy926细胞与THP-1细胞间黏附作用，降低EA.hy926细胞分泌的炎症因子IL-1、IL-6和IL-8水平。Western blot和RT-PCR结果表明蒙花苷能减少EA.hy926细胞中TLR4、NF-κB、ICAM-1、VCAM-1的蛋白表达量及ICAM-1、VCAM-1 mRNA的表达量，同时能升高细胞中IκBα的蛋白表达量。结论 蒙花苷能缓解血管内皮细胞炎症损伤，其作用机制可能主要是通过抑制TLR4/IκBα/NF-κB信号通路发挥效用。 |
| 关键词: 蒙花苷 TNF-α EA.hy926 炎症 IκBα/NF-κB |
| DOI：10.13748/j.cnki.issn1007-7693.2017.05.001 |
| 分类号:R284.1；R917.101 |
| 基金项目:国家自然科学基金（81274123）；浙江省重点实验室项目（2012E10002）；浙江省教育厅项目（Y201431439） |
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| Buddleoside Prevents TNF-α-induced Human Aortic Endothelial Cells Inflammatory Injury Through Inhibiting TLR4/IκBα/NF-κB Signaling Pathway |
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WU Yajun1, SU Jie1, HUANG Pujun2, CHEN Guoyang2, CHEN Suhong1,2,3, LYU Guiyuan1
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1.Zhejiang Chinese Medical University, Hangzhou 310053, China;2.Wenzhou Medical University, Wenzhou 325035, China;3.Zhejiang University of Technology, Hangzhou 310014, China
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| Abstract: |
| OBJECTIVE To study the protective effect and mechanism of buddleoside on human aortic endothelial cells. METHODS MTT assay was used to detect the viability of EA.hy926 in TNF-α and buddleoside. EA.hy926 cells were treated with TNF-α(50 ng·mL-1) for inflammatory injury model. After being pretreated with different concentrations(5, 15, 25 mmol·L-1) of buddleoside for 24 h, DAPI staining was used to evaluate the effect of buddleoside on adhesion of EA.hy926 and THP-1. The protein expression of TLR4, IκBα, NF-κB, ICAM-1 and VCAM-1 were tested by Western blot method respectively. The mRNA expression of ICAM-1 and VCAM-1 were tested by RT-PCR method respectively. ELISA method was used to measure the levels of IL-1, IL-6 and IL-8. RESULTS Buddleoside could inhibit the adhesion of TNF-α stimulated EA.hy926 cell and THP-1 cell. ELISA results showed that buddleoside reduced IL-1, IL-6 and IL-8 production in TNF-α-stimulated EA.hy926. Buddleoside significantly decreased TLR4, NF-κB, ICAM-1 and VCAM-1 protein expression in TNF-α-stimulated EA.hy926 cell. In addition, buddleoside increased IκBα protein expression. RT-PCR result showed buddleoside decreased ICAM-1 and VCAM-1 mRNA expression in TNF-α-stimulated EA.hy926 cell. CONCLUSION Buddleoside can alleviate the inflammatory injury of vascular endothelial cells, and its mechanism may be mainly through the inhibition of TLR4/IκBα/NF-κB signaling pathway. |
| Key words: buddleoside TNF-α EA.hy926 inflammation IκBα/NF-κB |
