血管紧张素Ⅱ-1型受体基因静默对肝枯否细胞NF-κB活性的影响  

Effect of AT-1 α receptor gene silencing on nuclear factor-κB activity in hepatic Kupffer cells

作　　者：李旭[1] 孟莹[2] 周高速[3] 张振书[4] 

机构地区：[1]南方医科大学南方医院急诊科,广东广州510515 [2]南方医科大学南方医院呼吸科,广东广州510515 [3]北京军区总医院急诊科,北京100700 [4]南方医科大学南方医院消化科,广东广州510515

出　　处：《南方医科大学学报》2009年第1期20-22,共3页Journal of Southern Medical University

基　　金：国家自然科学基金(30500243)

摘　　要：目的探讨肝枯否细胞血管紧张素Ⅱ1型受体(AT-1受体)基因静默对肝枯否细胞NF-κB信号通路的影响。方法采用原代分离培养的肝枯否细胞,免疫组化检测AT-1受体在肝枯否细胞的表达。构建pSilencer/AT-1α受体siRNA质粒,将其转染肝枯否细胞,予10-6mol/L血管紧张素Ⅱ(AngⅡ)刺激60min,设立阴性对照。凝胶电泳移动抑制实验检测NF-κBDNA结合活性。结果肝枯否细胞可以表达AT-1受体。AngⅡ可刺激肝枯否细胞NF-κBDNA结合活性增强;pSilencer/AT-1α受体siRNA转染细胞后可显著抑制AngⅡ诱导的NF-κBDNA结合活增强。结论AngⅡ可经肝枯否细胞AT-1α受体激活肝枯否细胞NF-κB活性。Objective To investigate the effect of angiotensin Ⅱ type-1 （AT-1）α receptor gene silencing on nuclear factor-κB （NF-κB） activity in hepatic Kupffer cells. Methods The expression of AT-1 α receptors in primary isolated cultured hepatic Kupffer cells was detected by immunohistochemistry, pSilencer/AT-1α receptor siRNA plasmids were transfected into Kupffer cells, which were subsequently exposed to 10^-6 mol/L angiotensin Ⅱ （Ang Ⅱ） for 60 min. The changes in the DNA binding activity of NF-κB in the cells was assessed using electrophoretic gel mobility shift assay （EMSA）. Results AT-1α receptor expression was detected in Kupffer cells. NF-κB DNA binding activity was markedly increased in Kupffer cells after Ang Ⅱ stimulation, and obviously inhibited by transfectiom with pSilencer/AT-1α receptor siRNA plasmid. Conclusion Ang Ⅱ stimulation of Kupffer cell results in increased activation of NF-κB via AT-1α receptor.

关 键 词：肝枯否细胞:血管紧张素Ⅱ1型受体 细胞粒因子-κB 

分 类 号：R575.2[医药卫生—消化系统]