Exosomes derived from umbilical cord‐derived mesenchymal stem cells exposed to diabetic microenvironment enhance M2 macrophage polarization and protect against diabetic nephropathy

Abstract The role of mesenchymal‐stem‐cell‐derived exosomes (MSCs‐Exo) in the regulation of macrophage polarization has been recognized in several diseases. There is emerging evidence that MSCs‐Exo partially prevent the progression of diabetic nephropathy (DN). This study aimed to investigate whether exosomes secreted by MSCs pre‐treated with a diabetic environment (Exo‐pre) have a more pronounced protective effect against DN by regulating the balance of macrophages. Exo‐pre and Exo‐Con were isolated from the culture medium of UC‐MSCs pre‐treated with a diabetic mimic environment and natural UC‐MSCs, respectively. Exo‐pre and Exo‐Con were injected into the tail veins of db/db mice three times a week for 6 weeks. Serum creatinine and serum urea nitrogen levels, the urinary protein/creatinine ratio, and histological staining were used to determine renal function and morphology. Macrophage phenotypes were analyzed by immunofluorescence, western blotting, and quantitative reverse transcription polymerase chain reaction. In vitro, lipopolysaccharide‐induced M1 macrophages were incubated separately with Exo‐Con and Exo‐pre. We performed microRNA (miRNA) sequencing to identify candidate miRNAs and predict their target genes. An miRNA inhibitor was used to confirm the role of miRNAs in macrophage modulation. Exo‐pre were more potent than Exo‐Con at alleviating DN. Exo‐pre administration significantly reduced the number of M1 macrophages and increased the number of M2 macrophages in the kidney compared to Exo‐Con administration. Parallel outcomes were observed in the co‐culture experiments. Moreover, miR‐486‐5p was distinctly expressed in Exo‐Con and Exo‐pre groups, and it played an important role in macrophage polarization by targeting PIK3R1 through the PI3K/Akt pathway. Reducing miR‐486‐5p levels in Exo‐pre abolished macrophage polarization modulation. Exo‐pre administration exhibited a superior effect on DN by remodeling the macrophage balance by shuttling miR‐486‐5p , which targets PIK3R1.