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目的:通过探讨HL-60细胞被TNF-α激活后NF-κB及IκBα活化的改变,了解银杏叶提取物(GbE)在治疗哮喘中部分可能的分子机理。方法:以HL-60细胞作为工具细胞,分別经TNF-α、GbE刺激,转染带有报告基因的质粒pNF-κB-Luc,通过荧光素酶的表达情况检测NF-κB活性变化;用免疫印迹方法检测IκBα的活化状况。结果:检测结果显示,以GbE预处理HL-60细胞可以明显抑制TNFα诱导的NF-κB转录激活的活性;但用GbE预处理细胞并不能拮抗TNF-α诱导的IκBα的磷酸化和降解,将GbE处理细胞时间延长和加大GbE的浓度同样不能阻止IκBα的磷酸化和降解。结论:GbE能抑制NF-κB激活的基因转录但并不是通过干扰经典的NIK/IKK/IκBα活化途径,而是通过干扰其它途径或采用其它方式来实现的。
OBJECTIVE: To investigate the possible molecular mechanism of Ginkgo biloba extract (GbE) in the treatment of asthma by exploring the activation of NF-κB and IκBα after HL-60 cells are activated by TNF-α. METHODS: HL-60 cells were used as the tool cells, stimulated with TNF-α and GbE, and transfected with the reporter gene plasmid pNF-κB-Luc. The activity of NF-κB was detected by luciferase expression. Blot method to detect the activation of IκBα. Results: The results showed that pretreatment of HL-60 cells with GbE could significantly inhibit the activity of NF-κB-induced transcriptional activation of TNF-α, but pretreatment with GbE did not antagonize the phosphorylation and degradation of IκBα induced by TNF-α. The prolongation of GbE-treated cells and increasing the concentration of GbE also failed to prevent the phosphorylation and degradation of IκBα. Conclusion: GbE can inhibit NF-κB-activated gene transcription but not by interfering with the classical NIK/IKK/IκBα activation pathway but by interfering with other pathways or using other approaches.