基础医学与临床 ›› 2015, Vol. 35 ›› Issue (8): 1020-1024.

• 研究论文 • 上一篇    下一篇

Vaspin对HUVEC中TNF-α介导的NF-κB和PI3K/Akt信号通路的影响

刘师伟1,王明明1,2,王军1,楼晓华3,董艳婷2,张丽1,赵术君1,何玉洁4   

  1. 1. 山西医科大学附属大医院
    2. 山西医科大学
    3. 美国休斯敦大学
    4. 山西医科大学第一医院
  • 收稿日期:2015-02-03 修回日期:2015-05-29 出版日期:2015-08-05 发布日期:2015-07-15
  • 通讯作者: 刘师伟 E-mail:lswspring6@aliyun.com
  • 基金资助:
    衰老调控因子SIRT6与NF-κB对话在人参皂苷Rg1延缓造血干/祖细胞衰老中的作用;衰老调控因子SIRT6与NF-κB对话在人参皂苷Rg1延缓造血干/祖细胞衰老中的作用;山西省留学回国人员科技活动择优资助项目

The effect of Vaspin on TNF-α-induced NF-κB and PI3K/Akt signaling pathways in HUVEC

  • Received:2015-02-03 Revised:2015-05-29 Online:2015-08-05 Published:2015-07-15
  • Contact: Shi-wei LIU E-mail:lswspring6@aliyun.com

摘要: 目的 探讨Vaspin对TNF-α介导的人脐静脉内皮细胞(HUVEC)NF-κB和PI3K/Akt信号通路的影响。方法 体外分离并培养HUVEC。NF-κB荧光酶报告质粒瞬时转染HUVEC,用不同浓度(0~320 μg/L)Vaspin预培养,随后用10 μg/L的TNF-α作用于HUVEC,荧光酶报告分析法测定NF-κB的转录活性。Western blot测定磷酸化的Akt水平。ELISA测定细胞上清液中IL-1及IL-6的浓度。Real time-PCR、Western blot检测细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)及单核细胞趋化蛋白-1(MCP-1)的mRNA和蛋白表达水平。结果 在HUVEC中,Vaspin抑制TNF-α介导的NF-κB转录活性(P<0.05),并且NF-κB下游因子IL-1、IL-6、ICAM-1 、VCAM-1 和MCP-1的表达降低(P<0.05)。Vaspin增加了炎性因子TNF-α介导的磷酸化的Akt水平(P<0.05)。结论 Vaspin抑制HUVEC中TNF-α介导的NF-κB信号通路,增强TNF-α介导的PI3K/Akt信号通路的传导。

关键词: Vaspin, 核因子-κB, 磷脂酰肌醇3激酶(PI3K)/Akt, 胰岛素抵抗, 炎症, 血管内皮细胞

Abstract: Objective To investigate the effects of visceral adipose tissue-derived serpin (Vaspin) on nuclear factor-kappa B (NF-κB) and phosphatidylinositol 3-kinase(PI3K)/Akt signal pathways in human umbilical vein endothelial cells (HUVEC) stimulated by tumor necrosis factor-α (TNF-α) to elucidate the role of Vaspin in human endothelial cells of inflammation and insulin resistance. Methods HUVEC were isolated and cultured in vitro. A NF-κB luciferase reporter system was constructed and transiently transfected into HUVEC. Following transfection, HUVEC were pretreated with various concentrations of Vaspin (0~320 μg/L) before 10 μg/L TNF-α stimulation. The transcription activity of NF-κB was determined using luciferase reporter assay. The level of Akt phosphorylation was checked by Western blot. Expression levels of NF-κB downstream inflammatory cytokines IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). mRNA and protein expression levels of 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 (qRT-PCR) and Western blot respectively. Results Vaspin inhibited TNF-α mediated activation of NF-κB and its downstream molecules (P<0.05). Vaspin significantly increased Akt phosphorylation in TNF-α stimulated endothelial cells (P<0.05). Conculsions Vaspin protected endothelial cells from TNF-α induced inflammation and insulin resistance by combination the inhibition of NF-κB, its downstream molecules and the upregulation of the PI3-kinase/Akt signal pathway.

Key words: Vaspin, NF-κB, phosphatidylinositol 3-kinase(PI3K)/Akt, insulin resistance, inflammation, endothelium

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