AT-1受体、ACE基因静默对肝星状细胞NF-κB活性的影响

Effect of angiotensin II type 1 receptor and angiotensin-converting enzyme gene silencing on nuclear factor-κB activity in hepatic stellate cells

目的探讨肝星状细胞血管紧张素Ⅱ1型(AT-1)受体及血管紧张素转换酶(ACE)基因静默对肝星状细胞NF-κB活性的影响。方法采用HSC-T6细胞系。构建pSilencer/AT-1α受体siRNA、pSilencer/ACE siRNA质粒,将其转染HSC-T6,予10-6mol/L血管紧张素Ⅱ(AngⅡ)或血管紧张素转换酶抑制剂(ACEI)干预,设立对照组。凝胶电泳移动抑制实验(EMSA)检测NF-κB DNA结合活性。结果EMSA显示:AngⅡ可刺激肝星状细胞NF-κB DNA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-κB DNA结合活性增强。pSilencer/ACE siRNA转染细胞后可抑制NF-κB DNA结合活性。结论AT-1受体、ACE基因静默可抑制肝星状细胞NF-κB的活性。

Objective To investigate the effect of angiotensin Ⅱ （AngⅡ） type 1 （AT-1） receptor and angiotensin-converting enzyme （ACE） gene silencing on nuclear factor-κB （NF-κB） activity in hepatic stcllatc cells （HSCs）. Methods pSilencer/AT-1 α receptor siRNA and pSilencer/ACE siRNA plasmids were transfected into cultured HSC-T6 cells, which were subsequently stimulated by 10-6 mol/L AngII or ACE inhibitor （ACEI）. The DNA binding activity of NF-κB in the transfected cells was analyzed using electrophoretic gel mobility shift assay （EMSA）. Results Gel shift studies showed that stimulation of the HSCs by AngII markedly increased the DNA-binding activity of NF-κB, which was inhibited by the transfection with pSilencer/ AT- 1 α receptor siRNA plasmid or pSilencer/ACE siRNA plasmid. Conclusion AT- 1 α receptor and ACE gene silencing result in inhibition of NF-κB activity in HSCs in vitro.