Journal of Shandong University (Health Sciences) ›› 2024, Vol. 62 ›› Issue (6): 17-29.doi: 10.6040/j.issn.1671-7554.0.2024.0163

• Preclinical Medicine • Previous Articles    

MicroRNA-210-3p inhibits inflammatory pain in rats by regulation ten-eleven translocation 2 expression

WEI Jiacheng1, YANG Baozhong2, WEI Wei1, XUE Yating1, CUI Chenlong1, FANG Jun1   

  1. 1. College of Anesthesiology, Shanxi Medical University, Taiyuan 030001, Shanxi, China;
    2. Department of Anaesthesia and Surgery, Taiyuan Central Hospital, Taiyuan 030006, Shanxi, China
  • Published:2024-07-15

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Abstract: Objective To investigate the roles and mutual regulatory mechanisms of microRNA-210-3p(miR-210-3p)and ten-eleven translocation 2(TET2)in a rat model of inflammatory pain induced by complete Freunds adjuvant(CFA). Methods Bioinformatics and dual-luciferase reporter assays were used to analyse and identify target genes regulated by miR-210-3p in rats. The combinations of plasmid and miR-210-3p cotransfection in the experiments were grouped as follows: pmirGLO+mimics-NC group, pmirGLO+mimics-miR-210-3p group, TET2-WT-pmirGLO+mimics-NC group, TET2-WT-pmirGLO+mimics-miR-210- 3p group, TET2-MT-pmirGLO+mimics-NC group and TET2-MT-pmirGLO+mimics-miR-210-3p group; 60 rats were divided into 4 groups according to the randomised numerical table method: Normal control(CON)group(n=20), Complete Freunds adjuvant(CFA)group(n=20 ), Complete Freunds adjuvant + adeno-associated virus vector negative control(CFA + AAV NC)group(n=10 ), Complete Freunds adjuvant + adeno-associated virus miR-210-3p inhibitor(CFA + AAVi)group(n=10 ). The rat inflammatory pain model was established by subcutaneous injection of CFA into the underside of the left hind paw; the intervention model was established by tail vein injection of AAV with miR-210-3p inhibitor; the behaviour of the rats was observed and measured; the expression of miR-210-3p was detected by RT-qPCR; Western blotting and immunofluorescence staining were used to detect changes in the expression level and fluorescence intensity of TET2 protein in the spinal cord of lumbar extension segments from L4 to L6; and immunofluorescence staining was used to observe the cellular expression localisation of TET2 protein in the rat spinal cord. Results Dual-luciferase assays confirmed a negative regulatory relationship between miR-210-3p and TET2, as evidenced by a binding site. CFA injection significantly decreased the mechanical paw withdrawal mechanical threshold(PWMT)and the thermal paw thermal withdrawal latency(PTWL)(P<0.05). An increase in miR-210-3p and a concomitant decrease in TET2 protein expression were observed in the CFA group(P<0.05). Immunofluorescence showed that TET2 protein mainly colocalised with neuronal cells and its expression in the spinal cord was decreased in the CFA group(P<0.05). After AAVi treatment, PWMT and PTWL were significantly higher in the CFA+AAVi group than in the CFA+AAV NC group(P<0.05), with increased TET2 protein levels(P<0.05). Conclusion miR-210-3p downregulates TET2 protein expression; its inhibition in rats with inflammatory pain significantly alleviates pain symptoms.

Key words: microRNA-210-3p, Ten-eleven translocation methylcytosine dioxygenase 2, Inflammatory pain, Complete Freunds adjuvant: Adeno-associated virus vector

CLC Number: 

  • R741
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