Upregulation of Spinal miR-155-5p Contributes to Mechanical Hyperalgesia by Promoting Inflammatory Activation of Microglia in Bone Cancer Pain Rats
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bone Cancer Pain Model
2.2. Behavioral Assessment
2.3. Tissue Preparation
2.4. Sequencing and Analysis of miRNAs
2.5. N9 Cell Culture and Treatment
2.6. Real-Time Quantitative PCR (RT-qPCR)
2.7. Western Blot Analysis
2.8. Fluorescence In Situ Hybridization (FISH) and Immunofluorescence Assay
2.9. Dual-Luciferase Reporter Assay
2.10. Lentivirus Infection In Vitro and In Vivo
2.11. Statistical Analysis
3. Results
3.1. Mechanical Hyperalgesia Developed in BCP Rats and Differentially Expressed miRNAs Were Filtered Out Using RNA-seq
3.2. miR-155-5p Could Be Expressed in the Cytoplasm of Microglia in the Lumbar Spinal Cord of BCP Rats
3.3. Microglia Were Dramatically Activated and Accompanied by the Increase in Inflammatory Cytokines in BCP Rats
3.4. The Expressions of miR-155-5p, Tnf, Il1b, and Il6 Were Increased in N9 Cells Treated with LPS
3.5. Knockdown of miR-155-5p Alleviated the Upregulation of Tnf, Il1b, and Il6 in LPS-Treated N9 Cells In Vitro
3.6. Sgk3 May Have Been a Direct Target of miR-155-5p In Vitro
3.7. Inhibiting miR-155-5p Altered the Expression of Sgk3, Alleviated Spinal Cord Inflammation, and Attenuated Mechanical Hyperalgesia in BCP Rats
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Jian, Y.; Song, Z.; Ding, Z.; Wang, J.; Wang, R.; Hou, X. Upregulation of Spinal miR-155-5p Contributes to Mechanical Hyperalgesia by Promoting Inflammatory Activation of Microglia in Bone Cancer Pain Rats. Life 2022, 12, 1349. https://doi.org/10.3390/life12091349
Jian Y, Song Z, Ding Z, Wang J, Wang R, Hou X. Upregulation of Spinal miR-155-5p Contributes to Mechanical Hyperalgesia by Promoting Inflammatory Activation of Microglia in Bone Cancer Pain Rats. Life. 2022; 12(9):1349. https://doi.org/10.3390/life12091349
Chicago/Turabian StyleJian, Yanping, Zongbin Song, Zhuofeng Ding, Jian Wang, Ruike Wang, and Xinran Hou. 2022. "Upregulation of Spinal miR-155-5p Contributes to Mechanical Hyperalgesia by Promoting Inflammatory Activation of Microglia in Bone Cancer Pain Rats" Life 12, no. 9: 1349. https://doi.org/10.3390/life12091349