Abstract
The current study tries to discuss the functional role of microRNA-497 (miR-497) in diabetic neuropathic pain (DNP) and the related downstream mechanism. Bioinformatics analysis was implemented for the identification of differentially expressed miRNAs and genes. DNP was simulated in rats through intraperitoneal injection of streptozotocin. The expression patterns of miR-497, USP15, NRF2, and G6PD were then determined. The binding of miR-497 and USP15 was confirmed. Using gain- and loss-of-function assays, we analyzed the critical role of miR-497-mediated USP15 in DNP through the NRF2/G6PD axis. Downregulated miR-497 and elevated USP15 were observed in the dorsal root ganglion neurons isolated from spinal cord tissues of STZ-induced DNP rats. miR-497 could alleviate DNP, which was associated with suppression of USP15, a confirmed target of miR-497. USP15 enhanced the degradation and ubiquitination of NRF2 and induced G6PD expression, leading to the progression of DNP. We highlighted the crucial role of miR-497-mediated USP15 in DNP through the NRF2/G6PD axis.
Graphical abstract
1. miR-497 is downregulated in DRG neurons from spinal cord tissues of STZ-induced DNP rats.
2. miR-497 inhibits the expression of USP15, thereby alleviating STZ-induced DNP in rats.
3. USP15 promotes ubiquitination and degradation of NRF2, reducing the expression of G6PD.
4. miR-497 alleviates STZ-induced DNP in rats by regulating the USP15/NRF2/G6PD axis.
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Data availability
The data that supports the findings of this study are available in the manuscript and supplementary materials.
Abbreviations
- DNP :
-
diabetic neuropathic pain
- DM :
-
diabetes mellitus
- T2DM :
-
type 2 diabetes mellitus
- T1DM :
-
type 1 diabetes mellitus
- miRNAs or miRs :
-
microRNAs
- USP15 :
-
ubiquitin-specific protease 15
- STZ :
-
streptozotocin
- TGF-β :
-
transforming growth factor-β
- Keap1 :
-
Kelch-like ECH-associated protein 1
- NRF2 :
-
nuclear factor-erythroid 2-related factor 2
- G6PD :
-
glucose-6-phosphate dehydrogenase
- ATCC :
-
American Type Culture Collection
- GEO :
-
Gene Expression Omnibus
- DMEM :
-
Dulbecco’s modified Eagle’s medium
- NC :
-
negative control
- cDNA :
-
complementary DNA
- ECL :
-
enhanced chemiluminescence
- FBS :
-
fetal bovine serum
- HRP :
-
horseradish peroxidase
- 3’-UTR :
-
3’-untranslated region
- MUT :
-
mutant
- oe :
-
overexpression
- PBS :
-
phosphate-buffered saline
- EP :
-
Eppendorf
- RT-qPCR :
-
reverse transcription-quantitative polymerase chain reaction
- DRG :
-
dorsal root ganglion
- GAPDH :
-
glyceraldehyde-3-phosphate dehydrogenase
- shRNA :
-
short hairpin RNA
- Co-IP :
-
co-immunoprecipitation
- TBST :
-
Tris-buffered saline Tween-20
- WT :
-
wild-type
- PWT :
-
paw withdraw threshold
- PWL :
-
paw withdraw latency
- MNCV :
-
motor nerve conduction velocity
- ANOVA :
-
analysis of variance
- USPs :
-
ubiquitin-specific peptidases
- NADPH :
-
nicotinamide adenine dinucleotide phosphate
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Tonghui Zhang and Ling Chen designed the study. Ling Wang and Ling Chen collated the data, carried out data analyses, and produced the initial draft of the manuscript. Tonghui Zhang and Ling Wang contributed to drafting the manuscript. All authors have read and approved the final submitted manuscript.
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The study was implemented following the ratification of the Ethics Committee of The First Affiliated Hospital of Jinzhou Medical University and in the light of the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health.
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Zhang, T., Wang, L. & Chen, L. Alleviative effect of microRNA-497 on diabetic neuropathic pain in rats in relation to decreased USP15. Cell Biol Toxicol 39, 1–16 (2023). https://doi.org/10.1007/s10565-022-09702-8
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DOI: https://doi.org/10.1007/s10565-022-09702-8