METHODS: Herein, we investigated the antiviral properties of IFN-α by introducing IFN-α expression plasmids into a well-established HBV Hydrodynamic Injection (HDI) mouse model and examined the impact of IFN-α or hepcidin treatment on macrophages derived from THP-1 cells. The cytokine profiles were analyzed using the cytometry microsphere microarray technology, and flow cytometry was used to analyze the polarization of macrophages. Additionally, the IL-6/JAK2/STAT3 signaling pathway and the hepcidin-ferroportin axis were analyzed to better understand the macrophage polarization mechanism.
RESULTS: As evidenced by the suppression of HBV replication, injection of an IFN-α expression plasmid and supernatants of IFN-α-treated macrophages exerted anti-HBV effects. The IFN-α treatment up-regulated IL-6 in mice with HBV replication, as well as in IFN-α-treated HepG2 cells and macrophages. Furthermore, JAK2/STAT3 signaling and hepcidin expression was promoted, inducing iron accumulation via the hepcidin-ferroportin axis, which caused the polarization of M1 macrophages. Furthermore, under the effect of IFN-α, IL-6 silencing or blockade downregulated the JAK2/STAT3 signaling pathway and hepcidin, implying that increased hepcidin expression under IFN-α treatment was dependent on the IL-6/JAK2/STAT3 pathway.
CONCLUSIONS: The IL-6/JAK2/STAT3 signaling pathway is activated by IFN-α which induces hepcidin expression. The resulting iron accumulation then induces the polarization of M1 macrophages via the hepcidin-ferroportin axis, yielding an immune response which exerts antiviral effects against HBV replication.
方法:这里,我们通过将IFN-α表达质粒引入建立良好的HBV流体动力学注射(HDI)小鼠模型来研究IFN-α的抗病毒特性,并检查IFN-α或铁调素治疗对THP-1细胞衍生的巨噬细胞的影响。使用细胞计数微球微阵列技术分析细胞因子谱,用流式细胞仪分析巨噬细胞的极化情况。此外,分析IL-6/JAK2/STAT3信号通路和铁调素-铁转运蛋白轴,以更好地了解巨噬细胞极化机制.
结果:HBV复制的抑制证明,注射IFN-α表达质粒和IFN-α处理的巨噬细胞上清液发挥抗HBV作用。IFN-α治疗上调小鼠HBV复制的IL-6,以及IFN-α处理的HepG2细胞和巨噬细胞。此外,JAK2/STAT3信号和铁调素表达被促进,通过铁调素-亚铁转运蛋白轴诱导铁积累,引起M1巨噬细胞的极化。此外,在IFN-α的作用下,IL-6沉默或阻断下调JAK2/STAT3信号通路和铁调素,这意味着IFN-α治疗下铁调素表达增加依赖于IL-6/JAK2/STAT3途径。
结论:IL-6/JAK2/STAT3信号通路被诱导铁调素表达的IFN-α激活。然后,所产生的铁积累通过铁调素-铁转运蛋白轴诱导M1巨噬细胞的极化,产生对HBV复制发挥抗病毒作用的免疫反应。