Vascular endothelial growth factor accelerates healing of foot ulcers in diabetic rats via promoting M2 macrophage polarization

IF 3.2 3区医学 Q2 ENDOCRINOLOGY & METABOLISM

Diabetic Medicine Pub Date : 2024-06-27 DOI:10.1111/dme.15388

Fei Liu, Xianrui Xu, Tao Sun

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引用次数: 0

Abstract

Aim

The objective was to investigate the specific role and the regulatory mechanism of vascular endothelial growth factor (VEGF) during wound healing in diabetic foot ulcer (DFU).

Methods

Streptozotocin-induced diabetic rats were used to establish a DFU animal model. VEGF and Axitinib (a specific inhibitor of VEGFR) were used for treatment in vivo. The wounds at different time points were imaged and histological analysis of the wounds were performed by haematoxylin and eosin (H&E) staining and Masson's trichrome staining. Immunohistochemical staining was conducted to examine CD31 and eNOS expression in the wounds. Immunofluorescence assay and quantitative real-time PCR were performed to examine macrophage markers. In addition, THP-1 was differentiated to macrophages, and then treated with interleukin (IL)-4 to induce M2 macrophages, followed by VEGF treatment. The conditional medium (CM) from VEGF-mediated macrophages were collected to culture human dermal fibroblasts (HDFs). Cell viability and migration were measured by Cell Counting Kit (CCK)-8, wound-healing and Transwell assays, respectively.

Results

VEGF treatment remarkably accelerated wound healing of DFU rats. VEGF promoted collagen deposition and elevated CD31 and eNOS expression, confirming the pro-angiogenesis of VEGF around diabetic wound in rats. Meanwhile, VEGF restricted pro-inflammatory cytokines and increased F4/80 and CD206 expression, highlighting the activated macrophages and enhanced M2 macrophages following VEGF treatment in diabetic wounds of DFU rats. However, Axitinib exerted an opposite function to VEGF in DFU rats. Moreover, VEGF directly promoted macrophage polarization toward M2 phenotype in vitro, and the CM from VEGF-mediated M2 macrophages markedly promoted HDFs proliferation, migration and collagen deposition.

Conclusion

VEGF might accelerate the wound healing of DFU through promoting M2 macrophage polarization and fibroblast migration.

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血管内皮生长因子通过促进 M2 巨噬细胞极化加速糖尿病大鼠足部溃疡的愈合。

目的：研究血管内皮生长因子（VEGF）在糖尿病足溃疡（DFU）伤口愈合过程中的特殊作用及其调控机制：方法：利用链脲佐菌素诱导的糖尿病大鼠建立糖尿病足溃疡动物模型。方法：用链脲佐菌素诱导的糖尿病大鼠建立 DFU 动物模型，使用血管内皮生长因子和阿昔替尼（血管内皮生长因子受体的特异性抑制剂）进行体内治疗。对不同时间点的伤口进行成像，并通过血栓素和伊红（H&E）染色和马森三色染色对伤口进行组织学分析。通过免疫组化染色检查伤口中 CD31 和 eNOS 的表达。免疫荧光检测和定量实时 PCR 检测巨噬细胞标记物。此外，将 THP-1 分化为巨噬细胞，然后用白细胞介素（IL）-4 诱导 M2 巨噬细胞，再用 VEGF 处理。收集血管内皮生长因子介导的巨噬细胞的条件培养基（CM）来培养人真皮成纤维细胞（HDFs）。细胞活力和迁移分别通过细胞计数试剂盒（CCK）-8、伤口愈合和 Transwell 试验进行测定：结果：VEGF 治疗显著加速了 DFU 大鼠的伤口愈合。结果：VEGF 能显著加速 DFU 大鼠伤口的愈合，促进胶原蛋白沉积，提高 CD31 和 eNOS 的表达，证实 VEGF 能促进糖尿病大鼠伤口周围的血管生成。同时，VEGF 限制了促炎细胞因子，增加了 F4/80 和 CD206 的表达，突出了 VEGF 处理 DFU 大鼠糖尿病伤口后巨噬细胞的活化和 M2 巨噬细胞的增强。然而，Axitinib 在 DFU 大鼠中发挥的作用与 VEGF 相反。此外，VEGF在体外直接促进巨噬细胞向M2表型极化，VEGF介导的M2巨噬细胞产生的CM明显促进了HDFs的增殖、迁移和胶原沉积：结论：VEGF可通过促进M2巨噬细胞极化和成纤维细胞迁移加速DFU的伤口愈合。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Diabetic Medicine 医学-内分泌学与代谢

CiteScore

7.20

自引率

5.70%

发文量

229

审稿时长

3-6 weeks

期刊介绍： Diabetic Medicine, the official journal of Diabetes UK, is published monthly simultaneously, in print and online editions. The journal publishes a range of key information on all clinical aspects of diabetes mellitus, ranging from human genetic studies through clinical physiology and trials to diabetes epidemiology. We do not publish original animal or cell culture studies unless they are part of a study of clinical diabetes involving humans. Categories of publication include research articles, reviews, editorials, commentaries, and correspondence. All material is peer-reviewed. We aim to disseminate knowledge about diabetes research with the goal of improving the management of people with diabetes. The journal therefore seeks to provide a forum for the exchange of ideas between clinicians and researchers worldwide. Topics covered are of importance to all healthcare professionals working with people with diabetes, whether in primary care or specialist services. Surplus generated from the sale of Diabetic Medicine is used by Diabetes UK to know diabetes better and fight diabetes more effectively on behalf of all people affected by and at risk of diabetes as well as their families and carers.”