Abstract：
Hepcidin is a crucial regulator of iron homeostasis with protective effects on liver fibrosis. Additionally, gut microbiota can also affect liver fibrosis and iron metabolism. Although the hepatoprotective potential of Akkermansia muciniphila and Faecalibacterium duncaniae, formerly known as F. prausnitzii, has been reported, however, their effects on hepcidin expression remain unknown. We investigated the direct and macrophage stimulation-mediated effects of active, heat-inactivated, and cell-free supernatant (CFS) forms of A. muciniphila and F. duncaniae on hepcidin expression in HepG2 cells by RT-qPCR analysis. Following stimulation of phorbol-12-myristate-13-acetate (PMA) -differentiated THP-1 cells with A. muciniphila and F. duncaniae, IL-6 concentration was assessed via ELISA. Additionally, the resulting supernatant was treated with HepG2 cells to evaluate the effect of macrophage stimulation on hepcidin gene expression. The expression of genes mediating iron absorption and export was also examined in HepG2 and Caco-2 cells via RT-qPCR. All forms of F. duncaniae increased hepcidin expression while active and heat-inactivated/CFS forms of A. muciniphila upregulated and downregulated its expression, respectively. Active, heat-inactivated, and CFS forms of A. muciniphila and F. duncaniae upregulated hepcidin expression, consistent with the elevation of IL-6 released from THP-1-stimulated cells as a macrophage stimulation effect in HepG2 cells. A. muciniphila and F. duncaniae in active, inactive, and CFS forms altered the expression of hepatocyte and intestinal iron-mediated absorption /exporter genes, namely dcytb and dmt1, and fpn in HepG2 and Caco-2 cells, respectively. In conclusion, A. muciniphila and F. duncaniae affect not only directly but also through macrophage stimulation the expression of hepcidin gene in HepG2 cells. These findings underscore the potential of A. muciniphila and F. duncaniae as a potential therapeutic target for liver fibrosis by modulating hepcidin and intestinal and hepatocyte iron metabolism mediated gene expression.
摘要：
铁调素是铁稳态的关键调节剂，对肝纤维化具有保护作用。此外,肠道菌群也会影响肝纤维化和铁代谢。尽管Akkermansia粘虫和duncaniae的肝保护潜力，原名F.Prausnitzii,据报道，然而,它们对hepcidin表达的影响仍然未知。我们研究了直接和巨噬细胞刺激介导的活性，热灭活，和无细胞上清液（CFS）形式的粘虫A.muciniphila和F.duncaniae对hepcidin在HepG2细胞中表达的RT-qPCR分析。用A.muciniphila和F.duncaniae刺激佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）分化的THP-1细胞后，通过ELISA评估IL-6浓度。此外,用HepG2细胞处理所得上清液以评估巨噬细胞刺激对hepcidin基因表达的影响。还通过RT-qPCR在HepG2和Caco-2细胞中检测了介导铁吸收和输出的基因的表达。Duncaniae的所有形式都增加了hepcidin的表达，而A.muciniphila的活性和热灭活/CFS形式上调和下调了其表达，分别。活动,热灭活，和CFS形式的A.muciniphila和F.duncaniae上调hepcidin表达，与THP-1刺激细胞释放的IL-6升高一致，作为HepG2细胞中的巨噬细胞刺激作用。A.粘液虫和F.duncaniae活跃，不活跃,和CFS形式改变了肝细胞和肠道铁介导的吸收/输出基因的表达，即HepG2和Caco-2细胞中的dcytb和dmt1和fpn，分别。总之,粘虫和隐球菌不仅直接影响HepG2细胞中hepcidin基因的表达，而且还通过巨噬细胞刺激影响HepG2细胞中hepcidin基因的表达。这些发现强调了A.muciniphila和F.duncaniae作为通过调节铁调素和肠和肝细胞铁代谢介导的基因表达的肝纤维化的潜在治疗靶标的潜力。
