miR-361-5p regulates SLC25A24 to maintain mitochondrial function and alleviate granulosa cell dysfunction in diminished ovarian reserve.

Abstract

Background: The aim of this study was to investigate the role of miR-361-5p (a tumor suppressor) in regulating granulosa cell function by targeting SLC25A24, a key mitochondrial protein, to uncover potential therapeutic targets for diminished ovarian reserve (DOR).

Methods: This study included patients undergoing assisted reproductive technology treatment at our hospital. Granulosa cells were isolated from follicular fluid, and KGN cells were used for in vitro experiments. miR-361-5p and SLC25A24 expression levels were manipulated using miRNA mimics and inhibitors, and their effects on cell viability, apoptosis, and mitochondrial function were assessed. Techniques employed included qRT-PCR, Western blot analysis, ELISA, JC-1 staining, and dual-luciferase reporter assays. Key quantitative metrics included changes in mitochondrial DNA (mtDNA), ATP production, and reactive oxygen species (ROS) levels.

Results: miR-361-5p expression was significantly lower in DOR patients' granulosa cells compared to controls (P < 0.01). miR-361-5p inhibition markedly decreased KGN cells viability and increased apoptosis (P < 0.01), while miR-361-5p overexpression had the opposite effects (P < 0.01). SLC25A24 expression was inversely correlated with miR-361-5p levels, and its knockdown reversed the effects of miR-361-5p inhibition. Additionally, miR-361-5p modulation significantly affected mitochondrial function, with its overexpression reducing ROS levels and increasing ATP production (P < 0.01).

Conclusion: miR-361-5p plays a pivotal role in maintaining mitochondrial function and reducing KGN cells dysfunction by targeting SLC25A24. These findings offer new insights into the molecular mechanisms of DOR and highlight miR-361-5p as a potential therapeutic target to enhance ovarian reserve and improve fertility outcomes.