【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。更多还原

【Abstract】 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-1 α 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-1 α 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-1 α receptor siRNA plasmid.Conclusion Ang II stimulation of Kupffer cell results in increased activation of NF-κB via AT-1 α receptor.更多还原