Objective: The effect of total flavonoids of litchi (TFL) on nuclear translocation of nuclear factor-kappa B (NF- kappa B) in rat hepatic stellate cell line (HSC-T6) induced by transforming growth factor - beta 1 (TGF- beta 1) in vitro was studied to explore the mechanism of action of anti-hepatic fibrosis drugs. Methods: HSC-T6 was cultured in vitro, induced by TGFβ1 for 24 h, and then treated with TFL at 125, 250 and 500 μg/ml for 48 h. The effect of TFL on NF-κB nuclear translocation in HSC-T6 was observed by confocal laser microscopy. The effects of TFL on the expression of TLR4, p-IκB ɑ, p-NF-κB p65, NF-κB and Collagen I protein were detected by western blot. The expressions of TLR4 and p-NF-κB p65 were detected by immunofluorescence. Data were presented as mean±SEM. Homogeneity test of variance was performed and then followed by one-way analysis of variance (ANOVA). The multiple comparisons between groups were performed by LSD test. P < 0.05 was considered statistically significant. Results: Confocal laser scanning microscopy showed TFL inhibited the nuclear translocation of NF-κB in activated HSC-T6 in a concentration-dependent manner and TFL down regulated the protein expression levels of TLR4, p-IκB ɑ, p-NF-κB p65, NF-κB and collagen I protein in HSC-T6 in a concentration-dependent manner. Conclusion: The mechanism of TFL against hepatic fibrosis may be related to the inhibition of nuclear translocation of NF-κb in the activated HSC-T6 and the expression of TLR4, P-iκbɑ, P-nf-κb p65, NF-κb and collagen I protein in HSC-T6.
目的： 研究荔枝核总黄酮（TFL）对体外由转化生长因子β1（TGFβ1）诱导活化的大鼠肝星状细胞T6细胞株（HSC-T6）中核因子κB（NF-κB）核转位及相关蛋白表达的影响，探究该药抗肝纤维化的作用机制。 方法： 体外培养HSC-T6，通过TGFβ1诱导24 h后以125、250、500 μg/ml的TFL干预48 h，采用激光共聚焦显微镜观察TFL对HSC-T6内NF-κB核转位的影响，并以Western blot法检测TFL对细胞中TLR4、p-IκB ɑ、p-NF-κB p65、NF-κB及CollagenⅠ蛋白表达水平的影响，免疫荧光法检测TLR4、p-NF-κB p65的表达。数据采用均数±标准差表示，先进行方差齐性检验，再行单因素方差分析（ANOVA），组间多重比较采用LSD检验，P<0.05为差异具有统计学意义。 结果： 激光共聚焦显微镜可见TFL抑制活化HSC-T6中NF-κB的核转位，并具有浓度依赖性。TFL可下调HSC-T6中TLR4、p-IκB ɑ、p-NF-κB p65、NF-κB及CollagenⅠ的蛋白表达水平，同时具有浓度依赖性。 结论： TFL的抗肝纤维化的机制可能与其抑制活化HSC-T6内NF-κB的核转位，下调HSC-T6中TLR4、p-IκB ɑ、p-NF-κB p65、NF-κB及CollagenⅠ的表达相关。.