microRNA-15b-5p shuttled by mesenchymal stem cell-derived extracellular vesicles protects podocytes from diabetic nephropathy via downregulation of VEGF/PDK4 axis

• Published in: Journal of bioenergetics and biomembranes 2022-02, Vol.54 (1), p.17-30
• Main Authors: Zhao, Tiantian, Jin, Qingsong, Kong, Lili, Zhang, Dongdong, Teng, Yaqin, Lin, Liangyan, Yao, Xiaoyan, Jin, Yongjun, Li, Minglong
• Format: Article
• Language: English
• Subjects: 3' Untranslated regions; Animal Anatomy; Animal Biochemistry; Animals; Apoptosis; Assaying; Binding; Biochemistry; Bioorganic Chemistry; Chemistry; Chemistry and Materials Science; Cholecystokinin; Chromosome 5; Diabetes; Diabetes mellitus; Diabetes Mellitus - metabolism; Diabetes Mellitus - pathology; Diabetic Nephropathies - genetics; Diabetic Nephropathies - metabolism; Diabetic Nephropathies - pathology; Diabetic nephropathy; Down-Regulation; End-stage renal disease; Extracellular vesicles; Extracellular Vesicles - metabolism; Extracellular Vesicles - pathology; Flow cytometry; Gene expression; Glucose - metabolism; Histology; Inflammation; Kidney diseases; Kinases; Mesenchymal stem cells; Mesenchymal Stem Cells - metabolism; Mesenchymal Stem Cells - pathology; Mice; MicroRNAs; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Morphology; Nephropathy; Organic Chemistry; Pathogenesis; Podocytes - metabolism; Podocytes - pathology; Protein Kinases; Pyruvic acid; Reporter gene; Ribonucleic acid; RNA; Stem cell transplantation; Stem cells; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism; Vesicles
• ISSN: 0145-479X; 1573-6881
• DOI: 10.1007/s10863-021-09919-y

Diabetic nephropathy (DN) is a severe complication of diabetes lethal for end-stage renal disease, with less treatment methodologies and uncertain pathogenesis. In the current study, we determined the role of mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) containing microRNA (miR)-15b-5p in DN. After extraction and identification of MSC-derived EVs, mouse podocyte line MPC5 was selected to establish an in vitro high-glucose (HG) cell model, where expression of miR-15b-5p, pyruvate dehydrogenase kinase 4 (PDK4) and VEGFA expression in tissues and cells were determined. The loss- and gain- function assays were conducted to determine the roles of miR-15b-5p, PDK4 and VEGFA. MPC5 cells were then co-cultured with MSC-derived EVs and their biological behaviors were detected by Western blot, CCK-8 assay, and flow cytometry. The binding relationship between miR-15b-5p and PDK43 by dual luciferase reporter gene assay. The expression of miR-15b-5p was downregulated in podocytes under HG environment, but highly expressed in mouse MSCs-derived EVs. EVs-derived miR-15b-5p could protect MPC5 cell apoptosis and inflammation. miR-15b-5p inhibited the expression of PDK4 by directly bound to the 3’UTR region of PDK4 gene. miR-15b-5p inhibits VEGF expression by binding to PDK4. Inhibition of PDK4 decreased VEGFA expression and reduced apoptosis and inflammation. Collectively, miR-15b-5p shuttled by MSC-derived EV can play protective roles in HG-induced mouse podocyte injury, possibly by targeting PDK4 and decreasing the VEGFA expression.