miR-6824-3p suppresses hepatitis B virus replication by targeting NRAS to regulate TNF-α secretion in macrophages
10.13303/j.cjbt.issn.1004-549x.2026.04.008
- VernacularTitle: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
- Author:
Simin LIN
1
;
Limin CHEN
1
;
Yujia LI
1
;
Shilin LI
1
Author Information
1. Institute of Blood Transfusion, Chinese Academy of Medical Sciences & Peking Union Medical College, Chengdu 610052, China
- Publication Type:Journal Article
- Keywords:
HBV;
miR-6824-3p;
macrophage;
TNF-α;
NRAS;
MAPK signalling pathway
- From:
Chinese Journal of Blood Transfusion
2026;39(4):465-477
- CountryChina
- Language:Chinese
-
Abstract:
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 immune regulatory mechanisms underlying persistent HBV infection. Methods: miR-6824-3p mimic and inhibitor were transfected into human THP-1-induced macrophages. Real-time quantitative PCR (qRT-PCR), enzyme-linked immunosorbent assay (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 target 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-associated 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). Conclusion: 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 modulating the host immune microenvironment.
