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基于Toll样受体4/核因子-κB信号通路研究莪术醇抗肝纤维化的分子机制
Molecular mechanism of the anti-liver fibrosis effect of curcumol: An analysis based on the TLR4/NF-κB signaling pathway
文章发布日期:2020年06月08日  来源:  作者:郑洋,王嘉孺,刘露露,等  点击次数:293次  下载次数:33次

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【摘要】:目的 研究莪术醇对肝窦内皮细胞TLR4/NF-κB信号通路的作用,以此来分析莪术醇抗肝纤维化的分子机制。方法 将50只清洁级KM小鼠随机分为空白组(n=10)、模型组(n=19)以及莪术醇组(n=21),除去建模过程中死亡的小鼠,最后每组各10只;细胞分为空白对照组、阳性对照组、莪术醇干预组以及PDTC(NF-κB特异性抑制剂)组。用HE和Masson染色观察各组小鼠肝脏结构的改变,用Western Blot和实时荧光定量PCR(RT-PCR)检测各组细胞TLR4/NF-κB信号通路上关键分子TLR4和NF-κB mRNA和蛋白的表达水平,用免疫荧光检测各组细胞NF-κB的表达及入核情况。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 RT-PCR检测发现莪术醇干预组TLR4 mRNA和NF-κB mRNA表达水平较阳性对照组下降,差异均有统计学意义(P值均<0.05);Western Blot检测发现莪术醇干预组TLR4和磷酸化NF-κB的表达水平较阳性对照组下降,差异均有统计学意义(P值均<0.05);免疫荧光检测发现莪术醇干预组NF-κB入核较阳性对照组明显改善。结论 莪术醇抑制TLR4/NF-κB信号通路的活动,可能是其抗肝纤维化作用机制之一。
【Abstract】:Objective To investigate the molecular mechanism of the anti-liver fibrosis effect of curcumol by observing the effect of curcumol on the TLR4/NF-κB signaling pathway in liver sinusoidal endothelial cells. Methods A total of 50 mice were randomly divided into blank group, model group, and curcumol group, and cells were divided into blank control group, LPS positive control group, curcumol intervention group, and PDTC group. HE staining and Masson staining were used to observe the change in liver structure; Western blot and quantitative real-time PCR (RT-PCR) were used to measure the protein and mRNA expression of the key molecules TLR4 and NF-κB in the TLR4/NF-κB signaling pathway; immunofluorescence assay was used to observe the expression and nuclear import of NF-κB in cells. A one-way analysis of variance was used for comparison between multiple  groups, and the least significant difference t-test was used for further comparison between two groups. Results RT-PCR showed that compared with the positive control group, the curcumol intervention group had significant reductions in the mRNA expression of TLR4 and NF-κB (both P<0.05). Western blot showed that compared with the positive control group, the curcumol intervention group had significant reductions in the expression of TLR4 and NF-κB (both P<0.05). Immunofluorescence assay showed that compared with the positive control group, the curcumol intervention group had significant improvement in NF-κB nuclear import. Conclusion Curcumol can exert an anti-liver fibrosis effect possibly by inhibiting the activity of the TLR4/NF-κB signaling pathway.
【关键字】:肝硬化; 莪术醇; 肝窦内皮细胞; Toll样受体4; NF-κB; 信号传导
【Key words】:liver cirrhosis; curcumol; liver sinusoidal endothelial cells; Toll-like receptor 4; NF-kappa B; signal transduction
【引证本文】:ZHENG Y, WANG JR, LIU LL, et al. Molecular mechanism of the anti-liver fibrosis effect of curcumol: An analysis based on the TLR4/NF-κB signaling pathway[J]. J Clin Hepatol, 2020, 36(7): 1508-1513. (in Chinese)
郑洋, 王嘉孺, 刘露露, 等. 基于Toll样受体4/核因子-κB信号通路研究莪术醇抗肝纤维化的分子机制[J]. 临床肝胆病杂志, 2020, 36(7): 1508-1513.

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