Injury - 2026-04-15 - Journal Article
Naringin targets JAK1-mediated M2 polarization of macrophages to promote the osteogenic effect of induced membrane technique.
Li S, Ye J, Yang D, Cai Q, Zeng Z, Zhou Q
Topics
Key Takeaway
Naringin accelerates induced membrane technique bone defect repair in rats by directly binding JAK1 to promote M2 macrophage polarization via the JAK1/STAT6 pathway, with si-JAK1 knockdown reversing this osteogenic effect.
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Summary
This study investigated whether Naringin enhances osteogenesis in the Masquelet (induced membrane technique) bone graft area by modulating macrophage polarization in a rat femoral critical-size defect model. RNA-seq identified JAK/STAT pathway upregulation; in vivo Naringin and IL-4 treatment increased M2 markers, osteogenic factor expression, and bone defect healing at 8 weeks. Molecular docking, MDS, and CETSA confirmed direct Naringin-JAK1 binding, and si-JAK1 knockdown reversed the pro-osteogenic effect in vitro.
Key Limitation
The study uses only a rat femoral defect model with no biomechanical endpoint, making it impossible to determine whether the observed increase in M2 polarization and osteogenic factor secretion translates to structurally competent bone regeneration.
Original 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.