›› 2010, Vol. 30 ›› Issue (4): 415-.

• 论著(基础研究) • 上一篇    下一篇

福辛普利拉对脂多糖诱导的单核细胞TLR4表达的抑制作用

李若谷, 方唯一, 曲新凯, 杨栓锁, 沙慧芳, 包国良   

  1. 上海交通大学 |胸科医院心内科, 上海 200030
  • 出版日期:2010-04-25 发布日期:2010-04-26
  • 通讯作者: 方唯一, 电子信箱: fwychest@163.com。
  • 作者简介:李若谷 (1976—), 主治医师, 博士生;电子信箱: pengyimin1977@163.com。

Inhibiting effect of Fosinoprilat on expression of Toll-like receptor 4 in lipopolysaccharide activated leucocythemia monocytic series cells

LI Ruo-gu, FANG Wei-yi, QU Xin-kai, YANG Shuan-suo, SHA Hui-fang, BAO Guo-liang   

  1. Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai 200030, China
  • Online:2010-04-25 Published:2010-04-26

摘要:

目的 研究福辛普利拉(Fos)对人白血病单核细胞系THP1 Toll样受体4(TLR4)表达的抑制作用。方法 培养THP1细胞,并将其浓度调整为1×106/mL,分为8组:对照组(正常THP1细胞)、脂多糖(LPS)刺激组(THP1细胞+ LPS)、DMSO组(THP1细胞+LPS+DMSO)及Fos干预组(THP1细胞+LPS+Fos,Fos质量浓度分别为0.25、0.5、1、5、10 μmol/L)。分别采用Real-time PCR法检测THP1细胞TLR4 mRNA的表达;Western blotting检测TLR4和NF-κB蛋白的表达;流式细胞仪检测TLR4蛋白的表达。结果 LPS刺激组THP1细胞TLR4 mRNA和蛋白的表达均高于对照组(P<0. 05);Fos干预组TLR4 mRNA和蛋白的表达均低于LPS刺激组, 其中1、0.5 μmol/L的Fos作用最明显。LPS刺激组NFκB蛋白表达明显高于各Fos干预组(P<0.05)。结论 Fos能有效下调LPS诱导的THP1细胞中TLR4的表达,并可有效抑制NF-κB的活性。

关键词: Toll样受体4, 福辛普利拉, 脂多糖, 白血病单核细胞系

Abstract:

Objective To investigate the inhibiting effect of Fosinoprilat (Fos) on expression of Toll-like receptor 4 (TLR4)in lipopolysaccharide (LPS) activated human leucocythemia monocytic series cells (THP1). Methods THP1 cells were cultured, and the concentration was adjusted to 1×106/mL. Eight groups were divided: control group (normal THP1 cells), LPS stimulation group (THP1 cells+LPS), DMSO group (THP1 cells+LPS+DMSO) and Fos intervention groups (THP1 cells+LPS+Fos, with Fos concentrations of 10,5,1,0.5,0.25 μmol/L, respectively). Real-time PCR was employed to detect the expression of TLR4 mRNA in THP1 cells, Western blotting was used to detected the expression of TLR4 and NF-κB protein, and the expression of TLR4 protein was detected by flow cytometry. Results The expression of TLR4 mRNA and protein of THP1 cells in LPS stimulation group was significantly higher than that in control group (P<0.05). The expression of TLR4 mRNA and protein of THP1 cells in Fos intervention groups (especially for 1 μmol/L Fos and 0.5 μmol/L Fos) was lower than that in LPS stimulation group. The expression of NF-κB protein in LPS stimulation group was significantly higher than that in Fos intervention groups (P<0.05). Conclusion Fos can downregulate the expression of TLR4 and inhibit the activity of NF-κB in LPS activated THP1 cells.

Key words: Toll-like receptor 4, Fosinoprilat, lipopolysaccharide, leucocythemia monocytic series