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
- 1 Department of Nephrology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), Hangzhou, Zhejiang 310000, China
- 2 Clinical School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang 310004, China
- 3 Department of Nephrology, Ningbo First Hospital, Ningbo, Zhejiang 315010, China
Received: October 13, 2023 Accepted: January 3, 2024 Published: February 8, 2024
https://doi.org/10.18632/aging.205535How to Cite
Copyright: © 2024 Jin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Objective: The exosomal cargo mainly comprises proteins, lipids, and microRNAs (miRNAs). Among these, miRNAs undertake multiple biological effects of exosomes (Exos). Some stem cell-derived exosomal miRNAs have shown the potential to treat diabetic nephropathy (DN). However, there is little research into the therapeutic effects of adipose-derived stem cell (ADSC)-derived exosomal miRNAs on DN. We aimed to explore the potential of miR-204-modified ADSC-derived Exos to mitigate DN.
Methods: Exos were extracted and identified from ADSCs. Histopathological injury, oxidative stress (OS), mitochondrial function, cell viability, and apoptosis were assessed to explore the effects of ADSC-derived Exos on DN. For mechanism exploration, quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to measure miR-204, methyltransferase (METTL3, METTL14, and METTL7A), and CIDEC. Also, CIDEC m6A methylation and miR-204-METTL7A, and METTL7A-CIDEC interactions were determined.
Results: Initially, OS-induced mitochondrial dysfunction was observed in DN rats. ADSC-derived Exos inhibited histopathological injury, cell apoptosis, OS, and mitochondrial dysfunction in DN rats. The similar therapeutic effects of ADSC-derived Exos were detected in the in vitro model. Intriguingly, miR-204 was released by ADSC-derived Exos and its upregulation enhanced the anti-DN effects of Exos. Mechanically, miR-204 reduced METTL7A expression to CIDEC m6A methylation, thus suppressing OS and mitochondrial dysfunction.
Conclusions: ADSC-derived exosomal miR-204 rescued OS-induced mitochondrial dysfunction by inhibiting METTL7A-mediated CIDEC m6A methylation. This study first revealed the significant role of ADSC-derived exosomal miR-204 in DN, paving the way for the development of novel therapeutic strategies to improve the clinical outcomes of DN patients.
Abbreviations
ATP: adenosine triphosphate; ADSC: adipose-derived stem cell; ACR: albumin-to-creatinine ratio; α-SMA: alpha-smooth muscle actin; BUN: blood urea nitrogen; CAT: catalase; CCK-8: cell counting kit-8; DN: Diabetic nephropathy; ESRD: end-stage renal disorder; ELISA: enzyme-linked immunosorbent assay; Exos: exosomes; FBG: fasting blood glucose; FBS: fetal bovine serum; FITC: fluorescein isothiocyanate; GPX: glutathione peroxidase; HE: hematoxylin-eosin; MDA: malondialdehyde; METTL: methyltransferase-like; MEM: Minimum Essential Medium; miR: miRNA; m6A: N6-methyladenosine; NAC: N-Acetylcysteine; OD: optical density; OS: oxidative stress; PAS: periodic acid-Schiff; PBS: phosphate-buffered saline; qRT-PCR: quantitative real-time polymerase chain reaction; ROS: reactive oxygen species; Scr: serum creatinine; SOD: superoxide dismutase; TC: total cholesterol; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; TGF: transforming growth factor; TEM: transmission electron microscopy; TG: triglyceride; UM: urinary microalbumin; WTAP: Wilms tumor 1-associated protein.