| BackgroundDiabetic skin ulcers are ulcers and gangrene that occur in diabetic patients due to microcirculatory disorders caused by neurovascular disease.It is often delayed and difficult to heal,and the life expectancy and quality of life significantly reduced.However,the causes of delayed wound healing or lack of healing in diabetes are still unclear,and there are still limited treatments that can accelerate or facilitate wound healing in diabetes.Endothelial cells as an important role in angiogenesis.During angiogenesis,vascular endothelial cells are rapidly activated and migrated to distant sites for proliferation.New primary capillaries are formed from existing capillaries to promote tissue repair.Endothelial dysfunction is the basis of diabetic microvascular and macrovascular complications.Exosomes and mi RNAs are important molecules involved in the pathogenesis of diabetes.These molecules can play their roles by targeting cellular and molecular pathways involved in different stages of the pathogenesis of diabetes.It has been reported that endothelial progenitor cells secrete exosomes to repair vascular endothelial injury,but the mechanism is not clear.Objective1.To investigate the expression profiles of mi RNA in endothelial progenitor cell exosomes and human umbilical vein exosomes.2.To clarify the repair effect of endothelial progenitor cell exosome and mi RNA-221-3p on diabetic skin injury.3.To explore the repair mechanism of mi RNA contained in endothelial progenitor cell exosome on diabetic skin injury.Method1.Cellular immunofluorescence assayThe induced endothelial progenitor cells were incubated with CD133 and Fl K-1antibodies,and then the second antibody was incubated and identified by fluorescence microscopy.2.Detection mi RNA of endothelial progenitor exosomes and human umbilical vein exosomes using high-throughput techniquesThe exosomes extracted from endothelial progenitor cells and human umbilical vein cells were used to construct mi RNAs library and sequenced on the computer.The original data were filtered and the corresponding software was used to qualitatively and quantitatively analyze the exosomes.3.Cell scratch testThe endothelial cells of thoracic aorta were treated with mi RNA-221-3p for 12 hours.The changes of cell movement and migration ability were observed under a microscope.4.Immunohistochemical experimentControl group and experimental group mice were sacrificed.The dermal tissue at the wound healing site was taken for immunohistochemistry and the expression of CD31,Ki67,VEGF and PCNA in the dermis was observed.Results1.Immunofluorescence results showed that both marker proteins CD133 and Fl K-1were positive,indicating that EPCs were successfully induced to differentiate.2.The results of high-throughput sequencing of mi RNAs from human umbilical vein endothelial cells and endothelial progenitor cell exosomes showed that there were significant differences in the expression profiles of mi RNAs between endothelial progenitor cell exosomes and human umbilical vein endothelial endothelial cell exosomes.The main functions of mi RNAs from endothelial progenitor cell exosomes were revealed through the analysis of the functions and signaling pathways.3.Animal experiment results showed that exosomes and mi RNA-221-3p could promot e wound healing in normal mice and diabetic mice.4.Immunohistochemical results showed that CD31,Ki67,VEGF and PCNA increased significantly and neovascularization increased in diabetic skin injury models i ncubated with endothelial progenitor exosomes and mi RNAs.5.The results of cell scratch test showed that mi RNA-221-3p could significantly enhan ce the migration ability of endothelial cells.Conclusion1.Endothelial progenitor cell exosomes and mi RNA-221-3p can promote wound healin g in diabetic mice,promote neovascularization and lead to the increase of CD31,Ki67,VEGF and PCNA proteins.2.mi RNA-221-3p can promote the migration of endothelial cells. |
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