Recombinant Human Fibronectin Mediates Macrophage Polarization via NF-κB/TGF-β1 Pathway to Enhance Fibroblast Proliferation

IF 2.4 4区生物学 Q4 CELL BIOLOGY

Biology of the Cell Pub Date : 2025-09-17 DOI:10.1111/boc.70025

Zongbao Zuo, Zizheng Yang, Jun Zhao, Lei Han, Jing Yang, Yueping Wang, Ziyun Zhang, Daoping Zhou

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引用次数: 0

Abstract

Objective

This research aims to explore the molecular mechanism where recombinant human fibronectin (rhFN) regulates macrophage polarization and then affects fibroblast proliferation via the nuclear factor kappa B (NF-κB)/transforming growth factor β1 (TGF-β1) signaling pathway.

Methods

Macrophages RAW 264.7 were activated with LPS and subsequently treated with rhFN, followed by flow cytometry to assess macrophag polarization. Cytokine levels of interleukin (IL)-10 tumor necrosis factor alpha (TNF-α), IL-6, and Arg-1, as well as TGF-β1, were measured using enzyme-linked immunosorbent assay (ELISA). Fibroblast NIH 3T3 was cultured with macrophage-conditioned media (CM), and CCK-8, cell adhesion, and wound healing assays were used to evaluate their proliferation, adhesion, and migration capacities. Western blot was conducted to detect the changes of proteins related to TGF-β1/Smad2/3 and NF-κB signaling.

Results

RhFN significantly promoted macrophage M2 polarization and increased TGF-β1 secretion while reducing pro-inflammatory cytokines TNF-α and IL-6, increasing IL-10 and Arg-1 levels. Fibroblasts cultured with rhFN-treated macrophage-CM showed increased Smad2/3 phosphorylation, causing improved proliferation, adhesion, and migration abilities. Inhibition of NF-κB signaling promoted an anti-inflammatory macrophage profile, while NF-κB activation partially reversed rhFN's effects on fibroblast function. Inhibition of TGF-β1 resulted in reduced fibroblast proliferation, adhesion, and migration abilities, confirming its pivotal role in rhFN-mediated effects.

Conclusion

RhFN modulates macrophage polarization through NF-κB inhibition and promotes fibroblast proliferation, adhesion, and migration via TGF-β1/Smad2/3 signaling.

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重组人纤维连接蛋白通过NF-κB/TGF-β1通路介导巨噬细胞极化促进成纤维细胞增殖

目的：本研究旨在探讨重组人纤维连接蛋白（rhFN）通过核因子κB (NF-κB)/转化生长因子β1 （TGF-β1）信号通路调控巨噬细胞极化进而影响成纤维细胞增殖的分子机制。方法：用LPS激活巨噬细胞RAW 264.7，然后用rhFN处理巨噬细胞，然后用流式细胞术评估巨噬细胞极化。采用酶联免疫吸附法（ELISA）检测白细胞介素(IL)-10、肿瘤坏死因子α (TNF-α)、IL-6、Arg-1及TGF-β1的细胞因子水平。用巨噬细胞条件培养基（CM）培养成纤维细胞NIH 3T3，用CCK-8、细胞粘附和伤口愈合试验评估其增殖、粘附和迁移能力。Western blot检测TGF-β1/Smad2/3、NF-κB信号通路相关蛋白的变化。结果：RhFN显著促进巨噬细胞M2极化，增加TGF-β1分泌，降低促炎细胞因子TNF-α、IL-6，升高IL-10、Arg-1水平。用rhfn处理的巨噬细胞- cm培养成纤维细胞显示Smad2/3磷酸化增加，导致增殖、粘附和迁移能力改善。NF-κB信号的抑制促进了抗炎巨噬细胞的分布，而NF-κB的激活部分逆转了rhFN对成纤维细胞功能的影响。抑制TGF-β1导致成纤维细胞增殖、粘附和迁移能力降低，证实了其在rhfn介导的作用中的关键作用。结论：RhFN通过抑制NF-κB调控巨噬细胞极化，并通过TGF-β1/Smad2/3信号通路促进成纤维细胞增殖、粘附和迁移。

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来源期刊

Biology of the Cell 生物-细胞生物学

CiteScore

5.30

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： The journal publishes original research articles and reviews on all aspects of cellular, molecular and structural biology, developmental biology, cell physiology and evolution. It will publish articles or reviews contributing to the understanding of the elementary biochemical and biophysical principles of live matter organization from the molecular, cellular and tissues scales and organisms. This includes contributions directed towards understanding biochemical and biophysical mechanisms, structure-function relationships with respect to basic cell and tissue functions, development, development/evolution relationship, morphogenesis, stem cell biology, cell biology of disease, plant cell biology, as well as contributions directed toward understanding integrated processes at the organelles, cell and tissue levels. Contributions using approaches such as high resolution imaging, live imaging, quantitative cell biology and integrated biology; as well as those using innovative genetic and epigenetic technologies, ex-vivo tissue engineering, cellular, tissue and integrated functional analysis, and quantitative biology and modeling to demonstrate original biological principles are encouraged.