肝复康对乙醛刺激肝星状细胞NF-кB通路活化的抑制作用

Effects of Gan-fu-kang on NF-кB signaling pathway in acetaldehyde treated hepatic stellate cells

目的分析核因子-кB(nuclear transcription factor-kappa B,NF-кB适通路在乙醛刺激肝星状细胞活化和肝复康调节中的变化,探究肝复康对乙醛刺激肝星状细胞NF-кB通路活化的抑制作用。方法将大鼠肝星状细胞株(HSC-T6)随机分为3组:正常对照组、乙醛刺激组及肝复康治疗组。免疫蛋白印迹法(Western-blot)在蛋白水平检测肝星状细胞中各组TNF-α的表达,逆转录-聚合酶链式反应(reverse transcriptase polymerase chain reaction,RT-PCR)在基因水平检测肝星状细胞各组TNF-α、IкB激酶IкK-α、NF-кBP65及NF-кB的抑制蛋白IкB-α的表达。结果 Western-blot结果显示:TNF-α在正常大鼠肝星状细胞中几乎不表达,而乙醛刺激组有显著阳性表达(P<0.01);肝复康治疗组TNF-α阳性表达则明显减少(P<0.05)。RT-PCR结果显示:与正常组相比,乙醛刺激组TNF-α、IкK-α、NF-кBP65的基因表达明显上调(P<0.01,P<0.05),IкB-α的表达降低(P<0.01),肝复康治疗组的各基因表达水平显著恢复。TNF-α和NF-кBP65的表达成正相关,和IкB-α的表达成负相关。结论 TNF-α介导的NF-кB信号转导通路在酒精性肝纤维化的发生和进展中呈活化状态,肝复康对其有显著性抑制作用。

Objective To explore the Effects of Gan-fu-kang（ GFK） on TNF-α mediated NF-кB signaling pathway in hepatic stellate cells. Methods Hepatic stellate cell（ HSC-T6） was randomly assigned to three groups： normal control group,acetaldehyde group and GFK group. The expression of TNF-α in HSCs was detected by Western blotting method. The m RNA expression of TNF-α,IKK-α,IкB-α,NF-кBP65 were observed by reverse transcriptase polymerase chain reaction（ RT-PCR）. Results Western blotting method assay for TNF-α protein expression： compared with the control group,the positive expression of TNF-α were increased significantly in the acetaldehyde group（ P〈0. 01）. While,the positive expression of TNF-α were decreased evidently in the GFK group（ P〈0. 05）. RT-PCR results showed： compared with the control group,the m RNA expression of TNF-α,IкK-αand NF-кBP65 in acetaldehyde group were increased（ P〈0. 05）. Above m RNA expression levels in GFK group were reduced significantly（ P〈0. 01）,in which the expression of IкB-α showed the opposite tendency. The expression of TNF-α and NF-кBp65 was positive correlation,and IкB-α was negative correlation. Conclusion TNF-α mediated NF-кB signaling pathway may be playing a promoting role in the occurrence and progression of liver fibrosis. The therapeutic effects of GFK may be involved with TNF-α mediated NF-кB signal transduction pathway.