黄芪皂苷Ⅲ抑制小鼠动脉平滑肌细胞TNFR1介导信号转导  

Inhibition of Astragaloside Ⅲ on TNFR1-mediated Signaling Pathways in Mouse Arterial Smooth Muscle Cells

作　　者：王海芳[1] 张琳萍[2] 霍雪萍[1] 赵向绒[1] 赵苗苗[1] 曹情雯 胡军[1] 刘勤社[4] 

机构地区：[1]陕西省人民医院中心实验室,陕西西安710068 [2]西安交通大学医学部,陕西西安710061 [3]陕西省中医医院急诊科,陕西西安710003 [4]陕西中医药大学,陕西西安712046

出　　处：《云南中医学院学报》2017年第3期1-6,共6页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81573823)

摘　　要：目的研究黄芪皂苷Ⅲ(astragaloside Ⅲ,ASⅢ)对小鼠动脉平滑肌细胞(arterial smooth muscle cells,ASMCs)中I型TNF-α受体(TNF-αreceptor type 1,TNFR1)介导信号通路的抑制作用,为中药黄芪的抗炎作用和抗动脉粥样硬化作用提供证据。方法以不同浓度ASⅢ预处理ASMCs,之后以TNF-α诱导细胞。采用WST-1实验检测细胞活力变化;采用实时定量RT-PCR技术检测细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)、凝集素样氧化低密度脂蛋白受体(lectin-like oxidized LDL receptor-1,Lox-1)和ATP结合盒转运蛋白A1(ATP-binding cassette transporter A1,ABCA1)m RNA表达变化;采用Western Blot方法检测ICAM-1、Lox-1和ABCA1蛋白表达变化,并检测ASⅢ对转录因子NF-κB(nuclear factor-κB)上游抑制蛋白IκBα(NF-κB inhibitor-α)、以及对信号分子AKT、细胞外受体活化激酶(extracellular receptor-activated kinases,ERKs)和p38磷酸化的影响。结果 (1)ASⅢ在30 n M^10μM范围内不抑制细胞活力;(2)ASⅢ剂量依赖性抑制TNF-α对ICAM-1、Lox-1和ABCA1 m RNA表达的调控;(3)ASⅢ剂量依赖性抑制TNF-α对ICAM-1、Lox-1和ABCA1蛋白表达的调控;(4)ASⅢ显著抑制TNFR1介导的IκBα磷酸化和降解;(5)ASⅢ显著抑制TNFR1介导的AKT、ERKs和p38磷酸化。结论 ASⅢ可全面抑制ASMCs中TNFR1介导信号通路激活,参与其抗炎和抗动脉粥样硬化作用。Objective To investigate the inhibitory effect of astragaloside Ⅲ （ASⅢ） on TNFR1-mediated signaling pathways in mouse arterial smooth muscle cells （ASMCs） and provide experimental evidence for the anti-inflammatory and anti-atherosclerotic actions of Chinese herbal medicine Radix Astragali （Huangqi）. Methods The effect of ASⅢ on the cell viability of ASMCs was evaluated by conducting a WST-1 assay. The mRNA levels of ICAM-1 （intercellular adhesion molecule-1）, Lox-1 （lectin-like oxidized LDL receptor-1） and ABCA1 （ATP-binding cassette transporter A1） in ASMCs were measured using quantitative RT-PCR technique. Western Blot was performed to detect the effect of ASⅢ on TNF-α-induced modulation of ICAM-1 , Lox-1 and ABCA1 protein levels. TNFR1-mediated phosphorylation and degradation of I Bα, and phosphorylation of AKT, ERKs and p38 were also measured by using Western Blot analysis. Results （1） ASⅢ, when used at a concentration between 30 nM and 10 μM, did not reduce the cell viability of ASMCs; （2） The TNFα-stimulated up-regulation of ICAM-1 and Lox-1 , and down-regulation of ABCA1 expression level were inhibited by ASⅢ in a concentration-dependent manner; （3） ASⅢ suppressed the TNFR1-mediated phosphorylation and degradation of IκBα protein; （4） ASⅢ suppressed TNFR1-mediated phosphorylation of AKT, ERKs and p38. Conclusion ASⅢ inhibits the TNFR1-mediated intracell ular signaling pathways in ASMCs , which may in part explain its anti-inflammatory and anti-antherosclerostic effects.

关 键 词：黄芪皂苷Ⅲ 动脉平滑肌细胞 肿瘤坏死因子-α TNFR1 NF—κB 抗炎作用 

分 类 号：R285.5[医药卫生—中药学]