Research Paper Volume 16, Issue 4 pp 3302—3331

Exosomes as nanostructures deliver miR-204 in alleviation of mitochondrial dysfunction in diabetic nephropathy through suppressing methyltransferase-like 7A-mediated CIDEC N6-methyladenosine methylation

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Figure 10. ADSC-derived Exos modified by miR-204 rescue OS-induced mitochondrial dysfunction in DN rats by inhibiting METTL7A-mediated CIDEC m6A methylation. (A, B) The levels of MDA, SOD, GPX, and CAT in the kidney tissues and plasma of rats in each group using ELISA. (C, D) The expression of Drp1, Fis1, OPA1, Mfn1, METTL7A, and CIDEC in the kidney tissues of rats in each group using western blotting. (E) CIDEC m6A methylation levels in HK-2 cells using MeRIP-qPCR. Rats in the Exo group were treated with 1.6 mg/kg ADSC-derived Exos via caudal vein injection every two days for eight successive weeks. Meanwhile, rats in the Exo-mimic NC and Exo-miR-204 mimic groups were treated with ADSC-derived Exos loaded with mimic-NC or miR-204 mimic, respectively. Rats in the DN group were injected with the same volume of saline. Data were expressed as mean ± standard deviation (n = 6/group). *p < 0.05 and **p < 0.01 vs. Control group; ^p < 0.05 and ^^p < 0.01 vs. DN + Exo-mimic NC group. Abbreviations: ADSC: adipose-derived stem cell; Exos: exosomes; miR-204: microRNA-204; OS: oxidative stress; DN: diabetic nephropathy; METTL: methyltransferase-like; CIDEC: cell death-inducing DFF45-like effector C; m6A: N6-methyladenosine; MDA: malondialdehyde; SOD: superoxide dismutase; GPX: glutathione peroxidase; CAT: catalase; ELISA: enzyme-linked immunosorbent assay; MeRIP-qPCR: methylated RNA immunoprecipitation-PCR.