木犀草素对肝星状细胞活化和CCl4诱导小鼠肝纤维化的抑制作用及机制研究

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摘要：

[目的]该研究为探讨木犀草素对肝星状细胞活化和CCl4诱导小鼠肝纤维化的抑制作用及其可能的作用机制.[方法](1)体外利用CCK-8及划痕实验检测木犀草素对人肝星状细胞LX-2的增殖及迁移的影响;(2)qRT-PCR及Western blot检测木犀草素对CollagenⅠ、α-SMA、Smad2和p-Smad2表达的影响;(3)体内利用CCl4诱导小鼠肝纤维化模型,天狼猩红染色观察小鼠肝组织纤维胶原表达的变化;(4)应用label-free蛋白定量技术分析蛋白质的差异表达情况.[结果](1)体外实验结果显示木犀草素对LX-2细胞增殖及迁移有显著抑制作用,并下调CollagenⅠ、α-SMA、p-Smad2的蛋白表达;(2)体内实验结果证明木犀草素可显著缓解CCl4诱导的肝纤维化的发病进程;(3)GO分析结果显示木犀草素所调控的差异蛋白主要涉及线粒体电子传递、氧化磷酸化等调节过程,KEGG通路分析显示差异蛋白主要参与Metabolic、ECM-receptor interaction、PI3K-Akt等信号通路.[结论]木犀草素通过下调LX-2细胞中TGF-β1/Smad2信号通路抑制纤维化进程,并可能还存在其他多种机制,未来深入研究此类调控机制将为木犀草素的临床应用提供支持.

[Objective]In order to investigate the effect and mechanism of luteolin on liver fibrosis in vitro and in vivo.[Methods](1)CCK-8 and scratch experiment was used to detect the actvity of LX-2 cells under normal conditions and TGF-β1 activation;(2)qRT-PCR and western blot detected the effects of luteolin on CollagenⅠ,α-SMA,Smad2 and p-Smad2 expression;(3)CCl_4-induced liver fibrosis in mice.Masson staining was used to observe the changes of fibrous collagen expression in mice before and after administration.(4)Label-free protein quantification was used to analyze differential expression of proteins.[Results](1)The vitro experiments showed that luteolin inhibited LX-2 proliferation,and significantly inhibited the expression of CollagenⅠ,α-SMA,Smad2,p-Smad2.(2)Animal experiments showed that luteolin could effectively improve CCl_4-induced liver injury and liver function damage.(3)GO analysis indicated down-regulated differential proteins mainly involved in mitochondrial electron transfer,oxidative phosphorylation regulation,KEGG pathway analysis showed that the differential proteins were mainly involved in Metabolic,ECM-receptor interaction,PI3K-Akt and other signaling pathways.[Conclusion]The result shows that luteolin can inhibits fibrosis of human hepatic stellate cells by inhibiting the TGF-β/smad2 signaling pathway.

作者：

刁莹王仲娟李敏

Diao Ying;Wang Zhongjuan;Li Min(Institutes of Biology and Medical Sciences,Soochow University,Suzhou 215123,China)

机构地区：

苏州大学生物医学研究院

出处：

《betway官方app 学报:自然科学版》 CAS 北大核心 2023年第5期131-137,I0011,I0012,共9页

Journal of Henan Normal University(Natural Science Edition)

基金：

国家自然科学基金(82171813)。

关键词：

木犀草素肝星状细胞 TGF-β1/Smad2 质谱