Li Jingyun, Zeng Siqi, Sun Yue, Zou Jijun, Zhang Enyuan, Yan Qiyue, Chen Ling, Li Jun
Nanjing Women and Children's Healthcare Institute, Women's Hospital of Nanjing Medical University (Nanjing Women and Children's Healthcare Hospital), 123rd Tianfei Street, Mochou Road, Nanjing 210004, China.
Department of Plastic&Cosmetic Surgery, Women's Hospital of Nanjing Medical University (Nanjing Women and Children's Healthcare Hospital), 123rd Tianfei Street, Mochou Road, Nanjing 210004, China.
Phytomedicine. 2025 Jul 25;143:156825. doi: 10.1016/j.phymed.2025.156825. Epub 2025 Apr 30.
Hypertrophic scarring represents a major clinical challenge worldwide, with current treatment strategies showing limited effectiveness. Gluconic acid (GLA), a naturally occurring glucose metabolite found in fruits, honey, kombucha tea, and wine, may provide new approach for scar treatment.
This study aimed to investigate the anti-scarring properties of GLA and underlying molecular mechanisms.
A comprehensive experimental study combined in vitro hypertrophic scar fibroblasts and in vivo rabbit ear scar model assays.
Hypertrophic scar fibroblasts were treated with GLA. Cell counting kit-8 (CCK-8) and flow cytometry were used to evaluate cell viability and apoptosis. The collagen and ACTA2 (actin alpha 2, smooth muscle) expressions were analyzed by qPCR and western blot. A rabbit ear scar model was applied to assess GLA's effects on scar formation and collagen deposition. Transcriptome sequencing, pull-down assays, western blotting and rescue experiments using AKT agonist SC79 were employed to identify GLA-regulated pathways. Molecular docking, pull-down, cellular thermal shift assays and co-localization studies were used to assess GLA's interaction with PLOD1 (procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1). E64d, MG132 and QX77 were added to analyze GLA's function mechanisms on PLOD1 protein expression. Autophagy activation was evaluated through autophagic flux assay, transmission electron microscopy and autophagy related protein expression analysis. Mitochondrial membrane potential was detected by JC-1 staining.
GLA suppresses collagen and ACTA2 expressions and exerted a mild inhibitory effect on cell proliferation or apoptosis in hypertrophic scar fibroblasts. And it diminishes scar formation and collagen content in the rabbit ear scar model. AKT (protein kinase B) and phosphorylated AKT (p-AKT) levels were significantly reduced after GLA treatment. Rescue experiments confirmed that GLA's effects are mediated through the AKT pathway. Moreover, GLA interacts with PLOD1, resulting in its autophagy-lysosomal degradation. Additionally, GLA treatment activated autophagy, reduced mTOR protein expressions, and had no significant effect on mitochondrial membrane depolarization, further contributing to its anti-scarring effects.
Our findings demonstrate that GLA attenuates hypertrophic scarring through multi-modal mechanisms involving PLOD1 targeting, AKT/mTOR pathway inhibition, and autophagy activation. This study provides both mechanistic insights and therapeutic potential for GLA in scar treatment.
肥厚性瘢痕是全球范围内一项重大的临床挑战，目前的治疗策略效果有限。葡萄糖酸（GLA）是一种天然存在的葡萄糖代谢产物，存在于水果、蜂蜜、康普茶和葡萄酒中，可能为瘢痕治疗提供新方法。
本研究旨在探究GLA的抗瘢痕特性及其潜在分子机制。
一项综合实验研究，结合体外肥厚性瘢痕成纤维细胞和体内兔耳瘢痕模型试验。
用GLA处理肥厚性瘢痕成纤维细胞。采用细胞计数试剂盒-8（CCK-8）和流式细胞术评估细胞活力和凋亡情况。通过qPCR和蛋白质免疫印迹分析胶原蛋白和ACTA2（肌动蛋白α2，平滑肌）的表达。应用兔耳瘢痕模型评估GLA对瘢痕形成和胶原蛋白沉积的影响。采用转录组测序、下拉试验、蛋白质免疫印迹以及使用AKT激动剂SC79进行的拯救实验来鉴定GLA调控的信号通路。运用分子对接、下拉试验、细胞热迁移试验和共定位研究来评估GLA与脯氨酰羟化酶1（PLOD1）的相互作用。加入E64d、MG132和QX77以分析GLA对PLOD1蛋白表达的作用机制。通过自噬通量试验、透射电子显微镜和自噬相关蛋白表达分析评估自噬激活情况。采用JC-1染色检测线粒体膜电位。
GLA抑制胶原蛋白和ACTA2的表达，并对肥厚性瘢痕成纤维细胞的细胞增殖或凋亡产生轻微抑制作用。它还能减少兔耳瘢痕模型中的瘢痕形成和胶原蛋白含量。GLA处理后，AKT（蛋白激酶B）和磷酸化AKT（p-AKT）水平显著降低。拯救实验证实GLA的作用是通过AKT信号通路介导的。此外，GLA与PLOD1相互作用，导致其通过自噬-溶酶体途径降解。此外，GLA处理激活了自噬，降低了mTOR蛋白表达，并且对线粒体膜去极化没有显著影响，这进一步促进了其抗瘢痕作用。
我们的研究结果表明，GLA通过多种机制减轻肥厚性瘢痕，这些机制包括靶向PLOD1、抑制AKT/mTOR信号通路和激活自噬。本研究为GLA在瘢痕治疗中的作用机制和治疗潜力提供了见解。
