Naringin targets JAK1-mediated M2 polarization of macrophages to promote the osteogenic effect of induced membrane technique
Injury. 2026 Apr 15;57(6):113290. doi: 10.1016/j.injury.2026.113290. Online ahead of print.
ABSTRACT
BACKGROUND: Induced membrane technique (IMT), a novel approach for reconstructing critical-size bone defect, encounters the challenge of lengthy mineralization time after bone grafting. This study is to explore the effect of Naringin on M2 macrophage polarization-mediated osteogenesis in the induced membrane's bone graft area.
METHODS: The IMT model was established in SD rats. After 8 weeks of treatment with Naringin and interleukin-4 (IL-4), the repair effect of femoral bone defects was evaluated. Meanwhile, RNA sequencing (RNA-seq) was performed on the bone tissue from rats treated with Naringin to detect changes in gene transcription levels. In vitro, Macrophages were divided into four groups: Control group, si-JAK1 + Naringin group, Naringin group and IL-4 group. At corresponding stages, cell proliferation, cell phenotype (M1 or M2), factors related to the JAK/STAT6 pathway, and osteogenic factors secreted by macrophages were assessed. Additionally, a macrophage-osteoblast coculture system was established to analyze the effects of osteogenic factors derived from M2 macrophages on osteoblasts' viability and mineralization.
RESULTS: The result of RNA-seq on the bone tissue in the bone graft area revealed that genes upregulated by Naringin were significantly enriched in biological processes related to immune regulation and the JAK/STAT pathway. The in vivo study indicated that there is an increase in markers of M1 macrophages and a decrease in markers of M2 macrophages in the bone grafting area of IMT. Treatment with Naringin and IL-4 could stimulate the polarization of M0 macrophages towards M2, accelerate the healing of bone defects, and increase expression of osteogenic factors and JAK1/STAT6 pathway factors. The in vitro experiments showed that treatment of primitive macrophages (M0) with Naringin and IL-4 led to an increase in the number of M2 macrophages, enhanced secretion of osteogenic factors, upregulation of the JAK1/STAT6 pathway. Conversely, the number of M1 macrophages decreased. Additionally, si-JAK1 was able to reverse the positive effect of Naringin on M2 macrophage polarization. Furthermore, after co-culturing macrophages and osteoblasts, it was found that osteogenic factors derived from polarization of M2 macrophages could stimulate the activity and mineralization of osteoblasts. Finally, Molecular docking, molecular dynamics simulation (MDS) and CETSA results indicated that Naringin can directly bind to JAK1 protein in macrophages and maintain its thermal stability.
CONCLUSIONS: JAK1-mediated polarization of M0 macrophages towards M2 has a positive regulatory function in osteoblasts' mineralization. Naringin targets JAK1 in macrophages within the IMT's bone graft area, maintaining its stability, promoting the activation and phosphorylation of the JAK1/STAT6 pathway, stimulating M2 polarization of macrophages, and thereby facilitating bone graft growth and accelerating the repair of large bone defects.
PMID:41997096 | DOI:10.1016/j.injury.2026.113290












