miR-6824-3p通过靶向NRAS调控巨噬细胞TNF-α分泌抑制乙型肝炎病毒复制OA

Objective To investigate the regulatory role of miR-6824-3p in macrophage function and its molecular mechanism in inhibiting hepatitis B virus(HBV)replication,thereby providing experimental evidence to elucidate the im-mune regulatory mechanisms underlying persistent HBV infection.Methods miR-6824-3p mimic and inhibitor were trans-fected into human THP-1-induced macrophages.Real-time quantitative PCR(qRT-PCR),enzyme-linked immunosorbent as-say(ELISA),neutral red uptake,reactive oxygen species(ROS)production,and fluorescent latex particle phagocytosis assays were employed to evaluate the effects of miR-6824-3p on macrophage phenotype and function.Through a combination of bioinformatics analysis,dual luciferase reporter assays,western blot,and siRNA interference techniques,we identified the tar-get gene of miR-6824-3p and examined their effects on downstream signaling pathways.qRT-PCR and western blot analyses were performed to assess the impact of miR-6824-3p-regulated macrophages on HBV DNA,pgRNA,cccDNA,and HBV-asso-ciated antigen levels in HepAD38 cells.Key effector molecules were identified through neutralization assays.Results miR-6824-3p mimic significantly promoted the expression and secretion of proinflammatory factors,such as TNF-α and IL-1β,in macrophages(P＜0.001),while concurrently reducing ROS production and phagocytosis(P＜0.05).Furthermore,miR-6824-3p downregulated NRAS expression in macrophages,which was accompanied by a reduction in MAPK signalling path-way activity(p-MEK,p-ERK).Compared to the control group,the medium of macrophages with overexpressed miR-6824-3p inhibited the expression of HBV DNA,pgRNA,cccDNA,and HBV-associated antigens HBsAg,HBeAg,and HBcAg in HepAD38 cells(P＜0.01).Similar results were also observed in the co-culture system of macrophages with HepAD38 cells.The addition of TNF-α neutralizing antibodies markedly attenuated the aforementioned antiviral effects(P＜0.001).Conclu-sion miR-6824-3p targets NRAS to affect the downstream MAPK signaling pathway,regulating the immune function of macrophages.The TNF-α induced by miR-6824-3p is one of the key molecules that suppress HBV replication.This study provides evidence for further elucidating the molecular mechanisms by which miRNAs influence HBV replication via modula-ting the host immune microenvironment.