Shang Ruo-Yu, Yang Jia-Cai, Hu Wen-Gang, Xiao Rong, Hu Dong-Sheng, Lin Zhi-Chen, Li Song, Wang Nan-Nan, Zheng Yin, Liu Zhi-Hui, Chen Yun-Xia, Wang Min-Jie, Wang Chao, Jiang Bo, Lin Guo-An, Li Xiao-Liang, Shang Xin-Zhi, Yan Tian-Tian, Luo Gao-Xing, He Wei-Feng
State Key Laboratory of Trauma and Chemical Poisoning, Institute of Burn Research, First Affiliated Hospital of Army Medical University (Third Military Medical University), Chongqing 400038, China; Chongqing Key Laboratory for Wound Repair and Tissue Regeneration, Chongqing 400038, China.
Department of Plastic and Burn Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400038, China.
Phytomedicine. 2025 May;140:156498. doi: 10.1016/j.phymed.2025.156498. Epub 2025 Feb 14.
Background Hypertrophic scarring is an abnormal condition involving excessive fibroblast activation, aberrant extracellular matrix deposition, and persistent inflammation. Current treatments have limited efficacy and potential adverse effects, necessitating the development of new approaches. Purpose In this study, we investigated the effects of artesunate (ART) on hypertrophic scar (HS) formation and explored the underlying cellular and molecular mechanisms. Methods ART was local injected in rabbit ear HS model to study its effect on HS formation. Cell viability was assessed using the CCK8 assay. Cell proliferation and targeted protein expression were detected by flow cytometry, immunofluorescence and immunohistochemistry staining. Scratch assays were performed to evaluate cell migration, while western blotting analysis was used to detect changes in protein expression. Results Local injection of ART significantly reduced scar protrusion and thickness, improved the immune microenvironment, and attenuated collagen deposition. ART suppressed fibroblast activation, endothelial-mesenchymal transition (EndMT), and angiogenesis in HS tissues. In vitro, ART inhibited TGF-β1-triggered fibroblasts activation and EndMT of human umbilical vein endothelial cells. Mechanistically, ART attenuated the activation of PI3K/AKT/mTOR and TGF-β/Smad pathways in both fibroblasts and human umbilical vein endothelial cells. Notably, the mTOR activator 740 Y-P reversed the fibrosis-inhibiting effects of ART in vitro and in vivo, highlighting the critical and intriguing role of PI3K/AKT/mTOR signaling in mediating the effects of ART. Furthermore, we first uncovered a crosstalk between PI3K/AKT/mTOR and TGF-β/Smad pathways, wherein PI3K/AKT/mTOR inactivation by ART partially contributed to the inhibition of TGF-β/Smad signaling. Conclusion In addition to fibroblast activation, our findings first demonstrate that ART effectively mitigates HS formation by modulating the immune microenvironment and inhibiting EndMT and fibroblast activation. These results provide new perspectives into the development of HS and underscore the promising potential of ART as a therapeutic option for debilitating condition.
肥厚性瘢痕是一种异常状况，涉及成纤维细胞过度活化、细胞外基质异常沉积和持续性炎症。目前的治疗方法疗效有限且存在潜在不良反应，因此需要开发新的治疗方法。目的：在本研究中，我们研究了青蒿琥酯（ART）对肥厚性瘢痕（HS）形成的影响，并探讨了其潜在的细胞和分子机制。方法：将ART局部注射到兔耳HS模型中，以研究其对HS形成的影响。使用CCK8法评估细胞活力。通过流式细胞术、免疫荧光和免疫组织化学染色检测细胞增殖和靶向蛋白表达。进行划痕试验以评估细胞迁移，同时使用蛋白质印迹分析检测蛋白表达变化。结果：局部注射ART可显著降低瘢痕突出度和厚度，改善免疫微环境，并减少胶原沉积。ART抑制HS组织中的成纤维细胞活化、内皮-间充质转化（EndMT）和血管生成。在体外，ART抑制TGF-β1诱导的成纤维细胞活化和人脐静脉内皮细胞的EndMT。机制上，ART减弱了成纤维细胞和人脐静脉内皮细胞中PI3K/AKT/mTOR和TGF-β/Smad信号通路的活化。值得注意的是，mTOR激活剂740 Y-P在体外和体内均逆转了ART的抗纤维化作用，突出了PI3K/AKT/mTOR信号在介导ART作用中的关键且有趣的作用。此外，我们首次发现PI3K/AKT/mTOR与TGF-β/Smad信号通路之间存在相互作用，其中ART导致的PI3K/AKT/mTOR失活部分促成了对TGF-β/Smad信号的抑制。结论：除了成纤维细胞活化外，我们的研究结果首次表明，ART通过调节免疫微环境、抑制EndMT和成纤维细胞活化，有效减轻HS形成。这些结果为HS的发展提供了新的视角，并强调了ART作为一种治疗衰弱性疾病的有前景的治疗选择的潜力。
