Human umbilical cord mesenchymal stem cell-derived exosomes promote microcirculation in aged diabetic mice by TGF-β1 signaling pathway

• Published in: Diabetology and metabolic syndrome 2023-11, Vol.15 (1), p.1-234, Article 234
• Main Authors: Fan, Weijian, Zhou, Mengdie, Zheng, Shaoqiu, Liu, Yang, Pan, Songsong, Guo, Peng, Xu, Minjie, Hu, Chao, Ding, Anle, Wang, Zan, Yin, Shiwu, Zuo, Keqiang, Xie, Xiaoyun
• Format: Article
• Language: English
• Subjects: Angiogenesis; Antibodies; Apoptosis; Biotechnology; Body weight; CD105; CD105 antigen; CD81 antigen; CD9 antigen; Cell proliferation; Centrifugation; Diabetes; Diabetes mellitus (non-insulin dependent); Diabetic peripheral angiopathy; Endothelial cells; Exosomes; Growth factors; Hyperglycemia; Immunofluorescence; Mesenchymal stem cells; Microvascular; Microvasculature; Molecular modelling; Morphology; Particle size; Permeability; Protein expression; Proteins; Proteomics; Signal transduction; Skeletal muscle; Stem cells; TGF-β1; Transforming growth factor-b1; Transmission electron microscopy; Umbilical cord
• ISSN: 1758-5996; 1758-5996
• DOI: 10.1186/s13098-023-01191-x

BackgroundMicrovascular dysfunction is one of the most common pathological characteristics in Type 2 diabetes. Human mesenchymal stem cell-derived exosomes (hUCMSCs-Exo) have diverse functions in improving microcirculation; however, the molecular mechanism of hUCMSCs-Exo in regulating burn-induced inflammation is not well understood.MethodshUCMSCs-Exo were extracted by hypervelocity centrifugation method, and exosome morphology was observed by transmission electron microscopy, exosome diameter distribution was detected by particle size analysis, and exosome specific proteins were identified by Western blot.2. DB/DB mice were randomly divided into exosomes group and PBS group. Exosomes and PBS were injected into the tail vein, respectively, and the calf muscle tissue was taken 28 days later. 0.5% Evans blue fluorescence assessment microvascular permeability. The expression of CD31 was detected by immunofluorescence.The morphology and function of microvessels in muscle tissue of lower limbs was evaluated by transmission electron microscopy.3. TMT proteomics was used to detect the changes of differential protein expression in lower limb muscle tissues of the PBS group and the exosome group, and data analysis was performed to screen key signal molecules and their involved biological pathways. Key signal molecules CD105 were verified by Western blot. The expression of TGF-β1 in exosomes were evaluated by Western blot.ResultsElectron microscopy showed that hUCMSCs-Exo presented a uniform vesicle structure, and NTA showed that its diameter was about 160 nm. Western blot showed positive expression of specific proteins CD9, CD81 and TSG101 on exosomes.2. There is no significant change in blood glucose and body weight before and after the exosome treatment. The exosome group can significantly reduce the exudation of Evans blue. Compared with the PBS group. Meanwhile, CD31 immunofluorescence showed that the red fluorescence of exosome treatment was significantly increased, which was higher than that of PBS group. Transmission electron microscopy showed smooth capillary lumen and smooth and complete surface of endothelial cells in the exosome group, while narrow capillary lumen and fingerlike protrusion of endothelial cells in the PBS group.3.Quantitative analysis of TMT proteomics showed that there were 82 differential proteins, including 49 down-regulated proteins and 33 up-regulated proteins. Go enrichment analysis showed that the differential proteins were involve