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Updated: Sep 29, 2025
Exploring the Regulation of Lipid Droplet Catabolism through Lipophagy
Published on: January 31, 2025
miR-296-5p promotes autophagy in mouse LS8 cells under excessive fluoride via AMPK/ULK1 pathways
Yinyue Luo1, Dongxin Da1, Qingqing Weng1
1Department of Preventive Dentistry, Shanghai Stomatological Hospital, Fudan University, China; Shanghai Key Laboratory of Craniomaxillofacial Development and Diseases, Fudan University, China.
Abstract:
Numerous microRNAs participate in regulating the pathological process of autophagy. We have found miR-296-5p is one of the most significantly down-regulated microRNAs in a high concentration of sodium fluoride. However, it is not clear whether miR-296-5p augments autophagy in dental fluorosis. Our purpose is to explore the function of miR-296-5p in regulating autophagy of excessive fluoride development. Thus, the cell line of ameloblasts LS8 was exposed to a 1.5 mM dose of NaF and miR-296-5p-mimics, Real-time qPCR, CCK-8 assays, Fluorescence imaging and Western blot analysis were performed. Autophagy was observed. As our results indicated, miR-296-5p overexpression in mouse LS8 cells significantly accelerated autophagy. The autophagy inhibition effect of miR-296-5p underexpression was consistent with the effect of the AMPK inhibitor. And we found that the expression of LC3II was decreased via down-regulation of AMPK. The change of ULK1 by miR-296-5p may be accomplished through AMPK. Thus, miR-296-5p may improve the secretion of autophagic mediators by activating AMPK/ULK1 expression in fluorosis, suggesting that miR-296-5p, AMPK/ULK1 may be potential therapeutic targets under the higher fluoride stimulation.
Insights
MicroRNA miR-296-5p accelerates autophagy in dental fluorosis by activating the AMPK/ULK1 pathway. This suggests miR-296-5p and AMPK/ULK1 are potential therapeutic targets for fluoride toxicity.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- MicroRNAs (miRNAs) are key regulators of cellular processes, including autophagy.
- Dental fluorosis is linked to excessive fluoride exposure, involving pathological changes in ameloblasts.
- miR-296-5p is significantly downregulated in high sodium fluoride conditions, but its role in fluorosis-related autophagy is unclear.
Purpose of the Study:
- To investigate the function of miR-296-5p in regulating autophagy during excessive fluoride exposure.
- To explore the potential therapeutic implications of miR-296-5p in dental fluorosis.
Main Methods:
- Ameloblast-like LS8 cells were treated with sodium fluoride (NaF) and miR-296-5p mimics.
- Techniques included Real-time quantitative PCR (qPCR), CCK-8 assays, fluorescence imaging, and Western blot analysis.
- Autophagy markers (e.g., LC3II, ULK1) and signaling pathways (AMPK) were assessed.
Main Results:
- Overexpression of miR-296-5p significantly accelerated autophagy in LS8 cells.
- miR-296-5p downregulation inhibited autophagy, mimicking the effect of an AMPK inhibitor.
- miR-296-5p appears to regulate ULK1 expression via the AMPK pathway, affecting LC3II levels.
Conclusions:
- miR-296-5p promotes autophagy in fluorosis by activating the AMPK/ULK1 signaling pathway.
- This miRNA may enhance the secretion of autophagic mediators under high fluoride conditions.
- miR-296-5p and the AMPK/ULK1 pathway represent potential therapeutic targets for managing dental fluorosis.
