HucMSC-Ex调控自噬增强高糖环境下真皮成纤维细胞功能

HucMSC-Ex Regulates Autophagy to Enhance the Function of Dermal Fibroblasts in a High-Glucose Environment

该研究旨在探讨在高糖损伤模型中,人脐带间充质干细胞源外泌体(human umbilical cord mesenchymal stem cell-derived exosome,hucMSC-Ex)对真皮成纤维细胞(dermal fibroblasts,DFs)的影响,并探究自噬在糖尿病足溃疡(diabetic foot ulcer,DFU)修复中的作用。通过多种方法(包括形态分析、粒度测量、表面标记表征等)对hucMSC和hucMSC-Ex进行全面的鉴定。同时,使用Western blot和免疫荧光分析鉴定原代DFs表面标志物。通过流式细胞术、qRT-PCR、活性氧(reactive oxygen species,ROS)检测以及线粒体膜电位的检测,分别评估高糖损伤模型中DFs的凋亡情况、炎症因子表达水平、活性氧产生情况和线粒体功能。为了深入研究自噬在修复DFU中的作用,该研究通过免疫组织化学染色和Western blot技术检测DFU患者组织中LC3BII/I的表达情况,并提取组织蛋白进行自噬相关蛋白的表达差异情况分析。此外,在使用自噬诱导剂和抑制剂后,对DFs给予hucMSC-Ex处理,观察其增殖和迁移情况。研究结果表明,hucMSC-Ex可能通过调控自噬反应来增强高糖损伤下DFs的功能,从而促进DFU的修复。

The objective of this study was to investigate the impact of hucMSC-Ex(human umbilical cord mesenchymal stem cell-derived exosome)on DFs(dermal fibroblasts)in a high-glucose damage model and to explore the role of autophagy in the repair of DFU(diabetic foot ulcer).A comprehensive identification of hucMSC and hucMSC-Ex was conducted using various methods,including morphological analysis,particle size measurement,and surface marker characterization.Additionally,surface markers of primary DFs were identified through Western blot and immunofluorescence analyses.The assessment of apoptosis,inflammatory factor expression,ROS(reactive oxygen species)production,and mitochondrial function in DFs under high-glucose damage conditions separately were performed using flow cytometry,qRT-PCR,ROS detection,and measurement of mitochondrial membrane potential.To further investigate the role of autophagy in DFU repair,the expression of LC3BII/I in DFU patient tissues was examined through immunohistochemistry and Western blot techniques.Differences in the expression of autophagy-related proteins were analyzed by extracting tissue proteins.Moreover,hucMSC-Ex was administered to DFs after treatment with autophagy inducers and inhibitors to observe their proliferation and migration capabilities.The research results suggest that hucMSC-Ex may enhance the function of DFs under high glucose damage by modulating the autophagic response,thereby promoting the repair of DFU.