Abstract
Background/aim
Metformin is a drug for treating type 2 diabetes mellitus (T2DM). Recently, metformin has been shown to reduce the risks of asthma-associated outcomes and asthma deterioration, thereby holding promise as a superior medicine for diabetic patients with asthma. However, the mechanism by which metformin reduces diabetic asthma is yet to be clarified. This study aimed at ascertaining the downstream molecules underlying the effect of metformin on the activation of mast cells (MCs) and airway reactivity in a concomitant diabetic and asthmatic rat model.
Methods
A T2DM model was induced utilizing a high-fat diet and streptozotocin. Then, 10% ovalbumin was utilized to stimulate asthma-like pathology in the T2DM rats. RBL-2H3 cells were induced by anti-dinitrophenyl-specific immunoglobulin E for constructing an in vitro model. Luciferase assay and RNA immunoprecipitation (IP) assay were conducted to identify the interaction between microRNA-152-3p (miR-152-3p) and DNA methyltransferase 1 (DNMT1), while chromatin IP to identify the binding of DNMT1 to insulin receptor (IR) and insulin-like growth factor 1 receptor (IGF-1R) promoters. The effects of metformin on both pathological changes in vivo and biological behaviors of cells were evaluated. Using gain- and loss-of-function approaches, we assessed the role of the two interactions in the metformin-induced effect.
Results
It was suggested that metformin could impede the MC activation and airway resistance in the concomitant diabetic and asthmatic rats. Additionally, metformin downregulated IR and IGF-1R through DNMT1-dependent methylation to repress MC activation and airway resistance. DNMT1 was testified to be a target gene of miR-152-3p. Furthermore, miR-152-3p-induced silencing of DNMT1 was blocked by metformin, hence restraining MC activation and airway resistance.
Conclusion
The findings cumulatively demonstrate that metformin downregulates IR/IGF-1R to block MC activation and airway resistance via impairing the binding affinity between miR-152-3p and DNMT1.
Graphical Abstract
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Data availability
The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.
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Funding
This research was funded by the grants from the National Natural Science Foundation of China (No. 81860239) and Guangxi Provincial Natural Science Foundation (No. AD19110125).
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HL and FD conceived the ideas. HL and FD designed the experiments. FD, ZHL, HY, LJJ, FHF, and LAG performed the experiments. ZHL, HY, LJJ, FHF, and LYF analyzed the data. FD and ZHL provided critical materials. FHF, LAG, and LYF wrote the manuscript. HL supervised the study. All the authors have read and approved the final version for publication.
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All experimental procedures were performed based on the Guide for the Care and Use of Laboratory Animals (National Institutes of Health) and the animal experimental protocol approved by the Animal Ethics Committee of Guilin Medical University (GLMC202203061).
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Graphical headlights
Metformin reduces IR and IGF-1R in diabetic rats with asthma by regulating DNMT1.
miR-152-3p can target DNMT1.
DNMT1 enhances the methylation levels of IR and IGF-1R.
Metformin represses MC activation via regulating the miR-152-3p/DNMT1/IR/IGF-1R axis.
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Fu, D., Zhao, H., Huang, Y. et al. Metformin regulates the effects of IR and IGF-1R methylation on mast cell activation and airway reactivity in diabetic rats with asthma through miR-152-3p/DNMT1 axis. Cell Biol Toxicol 39, 1851–1872 (2023). https://doi.org/10.1007/s10565-022-09787-1
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DOI: https://doi.org/10.1007/s10565-022-09787-1