Formononetin enhances angiogenesis in diabetic wounds by inhibiting ferroptosis through suppression of mtROS-mediated xCT/GPX4 upregulation

IF 2.5 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-09-18 DOI:10.1016/j.tice.2025.103141

Xinying Li , Ding Zhu , Yanguo Wang, Chuanqi Zhao, Yuangang Lu

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引用次数: 0

Abstract

Background

Impaired angiogenesis critically contributes to delayed wound healing in diabetic patients. Mitochondrial damage in endothelial cells worsens this vascular dysfunction. Formononetin (FMN), a phytoestrogen from Astragalus roots, is traditionally used to enhance blood function and microcirculation; however, its mechanism remains unclear. This study investigates whether FMN protects human umbilical vein endothelial cells (HUVECs) from high glucose (HG)-induced ferroptosis, and we also evaluated its potential to enhance vascular repair and accelerate diabetic wound healing.

Methods

We used Western Blotting to measure ferroptosis markers including the cystine/gluta-mate antiporter system (xCT) and glutathione peroxidase 4 (GPX4) in both HUVECs and diabetic mouse wound tissue. To assess oxidative stress, we measured glutathione content (GSH), malondialdehyde production (MDA), and intracellular iron accumulation. Mitochondrial changes were evaluated through MitoSOX, TOMM20 protein expression, and JC-1 fluorescence. Wound tissue sections were processed for H&E staining to examine healing stages, and CD31 immunofluorescence was performed to visualize angiogenesis in the wound tissue.

Results

FMN effectively reduced ferroptosis markers in HG-treated HUVECs, and Erastin treatment abolished this protective effect. The compound appeared to block ferroptosis through two mechanisms: restoration of mitochondrial integrity and reactivation of the xCT/GPX4 antioxidant system. When we tested FMN in diabetic mice, wound closure rates improved substantially, the expression of xCT and GPX4 was increased, and CD31 expression in wound vessels increased, which matched what we observed in vitro.

Conclusion

Ferroptosis is critically involved in the pathogenesis of diabetic wounds. FMN exerts protective effects against HG-induced ferroptosis in HUVECs via restoration of mitochondrial function and mitigation of mitochondrial reactive oxygen species (mtROS) accumulation through xCT/GPX4 activation. This evidence suggests FMN could serve as a promising natural therapeutic agent for diabetic wound treatment.

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刺芒柄花素通过抑制mtros介导的xCT/GPX4上调来抑制铁下垂，从而促进糖尿病创口血管生成

背景：糖尿病患者的血管生成受损是导致伤口延迟愈合的关键因素。内皮细胞的线粒体损伤加重了这种血管功能障碍。刺芒柄花素（FMN）是一种来自黄芪根的植物雌激素，传统上用于增强血液功能和微循环；然而，其机制尚不清楚。本研究探讨了FMN是否能保护人脐静脉内皮细胞（HUVECs）免受高糖（HG）诱导的铁凋亡，并评估了其增强血管修复和加速糖尿病伤口愈合的潜力。方法采用Western Blotting检测HUVECs和糖尿病小鼠伤口组织中胱氨酸/谷胱甘肽反转运系统（xCT）和谷胱甘肽过氧化物酶4 （GPX4）的凋亡标志物。为了评估氧化应激，我们测量了谷胱甘肽含量（GSH）、丙二醛产量（MDA）和细胞内铁积累。通过MitoSOX、TOMM20蛋白表达和JC-1荧光检测线粒体变化。创面组织切片进行H&；E染色检查愈合阶段，CD31免疫荧光观察创面组织血管生成情况。结果fmn有效降低hg处理的huvec中铁凋亡标志物，而Erastin处理则消除了这种保护作用。该化合物似乎通过两种机制阻止铁凋亡：恢复线粒体完整性和重新激活xCT/GPX4抗氧化系统。当我们在糖尿病小鼠中测试FMN时，伤口愈合率明显提高，xCT和GPX4的表达增加，伤口血管中CD31的表达增加，与我们在体外观察到的结果一致。结论上睑下垂在糖尿病创面的发病机制中起重要作用。FMN通过激活xCT/GPX4，恢复线粒体功能，减少线粒体活性氧（mtROS）积累，对hg诱导的HUVECs铁凋亡具有保护作用。这表明FMN可以作为一种很有前途的天然治疗剂用于糖尿病伤口治疗。

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

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.