Effect Of Age Factors On APC-exos Ameliorate The Aging Of Human Skin Fibroblasts And Its Mechanisms

Objective:To study the effects of age factors on the adipose progenitor cells proliferation and biological characteristics of exosomes(e.g.exosomes morphology,particle size distribution,exosomes markers,etc.);APC-exos Ameliorate the Aging of Human Skin Fibroblasts and its mechanisms;To compare the effects of age factors:to compare the effects APC-exo anti-aging of skin fibroblasts from different age groups.Methods:A total of 17 cases of subcutaneous fat were collected in this study.Sample donors were assigned to two groups according to their age:Young(18–25 years),Old(60–67years).the adipose vascular matrix components(SVF)were first obtained by enzymatic digestion,and then the adipose progenitor cells(adipose promoter cell,APC)were selected by magnetic sorting.then,Compare the proliferation capacity of the two groups of adipocytes.Two groups of APC were expanded to collect culture medium to obtain exosomes by differential centrifugation.the obtained two groups of exosomes were compared for particle size;electron microscopy;western blot detection APC-exos membrane surface iconic protein CD63?TSG101?CD9 etc.The aging model of skin fibroblasts was established by hydrogen peroxide induced method,and the adipose progenitors of different age groups were co-cultured with aging skin fibroblasts.The related indexes of aging(p21,p53,sirt1)were detected by western blot,immunofluorescence staining,beta-galactosidase staining to detect the improvement of aging and the content of reactive oxygen species(ROS)in cells.Study on signaling pathway,pathway analysis:The aging model of skin fibroblasts was induced by hydrogen peroxide(H₂O₂),and the adipose-derived progenitor cells were cultured together with the aged skin fibroblasts in four groups.(1)normal skin fibroblasts treated without hydrogen peroxide;(2)aging skin fibroblasts treated with pbs;(3)aging skin fibroblasts treated with o APC-Exos;(4)aging skin fibroblasts treated with y APC-Exos.To explore the real effect of fat age factors on skin aging,to explore the composition differences of exosomes bodies of fat progenitor cells in different age groups,and to analyze the reasons.Results:1.The proliferation ability of adipose progenitor cells in the younger group was better than the older group.2.Human umbilical vein endothelial cells(huvecs)were cultured with different age groups of APC exosomes,the ability of huvec tube and migration was enhanced(p<0.05).In the younger group,adipose-derived exosomes increased the function of human umbilical vein endothelial cells(p<0.05).3.The results of transwell experiment showed that the migration ability of human skin fibroblasts was increased after co-culture with exosomes from young and old H-APC,Moreover,the migratory capacity of HDFs was significantly greater in the y APC-Exo group than in the o APC-Exos group(p<0.05).4.The normal skin fibroblasts induced by hydrogen peroxide showed significant senescence characterization including?-galactosidase staining positive and(p21?p53?sirt1)changes(p<0.05).5.Senescent cell were co-cultured with different age groups of APC exosomes,Signs of ageing reversed,Moreover,y APC-Exo was significantly greater of than the o APC-Exos.6.Exosomes of adipose progenitor cells reduces ROS levels of senescent cells Conclusion The exosomes derived from adipose progenitor cells has some characteristics of progenitor cells and plays an important role in wound repair and anti-aging and so on.The results show that the exosomes of adipose progenitor cells can enhance the function of fibroblasts and human umbilical vein endothelial cells,moreover,The exosomes derived from Adipose progenitor cells can reverse the aging of skin fibroblasts by acting on the p53/p21 signaling pathways.More important,The results showed that the exosomes derived from the young group was more effective than the old group.which may provide a potential new therapeutic idea for reversing skin aging.