Electrical stimulation induces miR-741-3p to regulate Radil and promote Schwann cell migration

doi:10.12307/2025.080

Abstract

Abstract: BACKGROUND: More and more animal experiments and clinical studies have confirmed that electrical stimulation can promote the repair of peripheral nerve injury, but the specific mechanism is not yet fully understood.
OBJECTIVE: To investigate the effect of electrical stimulation-induced miR-741-3p regulating Radil on Schwann cell migration.
METHODS: (1) Twelve male SD rats were randomly divided into electrical stimulation group and control group. The electrical stimulation group received continuous electrical stimulation for 7 days after sciatic nerve compression injury, while the control group was not treated after sciatic nerve compression. The injured nerves were taken on day 7 after operation. The expression difference of miR-741-3p between the two groups was verified by fluorescence in situ hybridization. (2) The target genes of miR-741-3p were predicted by miRDB, TargetScan, and miRWalk databases. (3) Schwann cells were transfected with miR-741-3p mimetic and its control, miR-741-3p inhibitor and its control, Radil siRNA and its control, miR-741-3p inhibitor+Radil siRNA and miR-741-3p inhibitor+siRNA control. The transfection efficiency was detected by RT-PCR. The migration ability of Schwann cells was detected by Transwell chamber.
RESULTS AND CONCLUSION: (1) The fluorescence intensity of miR-741-3p in the electrical stimulation group was lower than that in the control group. (2) The results of database prediction showed that 69 genes might be the target genes of miR-741-3p. Radil was one of the predicted target genes, which was mainly involved in cell adhesion and migration. (3) Compared with the miR-741-3p inhibitor control group, the number of Schwann cell migration increased in the miR-741-3p inhibitor group (P < 0.05). Compared with the miR-741-3p mimic control group, the number of Schwann cell migration in the miR-741-3p mimic group decreased (P < 0.05). Compared with the siRNA control group, the number of Schwann cell migration was decreased in the Radil siRNA group (P < 0.05). (4) Compared with miR-741-3p inhibitor control group, the expression level of Radil was increased in miR-741-3p inhibitor group. Compared with miR-741-3p mimic control group, the expression level of Radil was decreased in miR-741-3p mimic group. (5) Compared with miR-741-3p inhibitor+siRNA control group, the number of Schwann cell migration was reduced in miR-741-3p inhibitor+Radil siRNA group (P < 0.05). The results showed that electrical stimulation promoted the migration of Schwann cells by down-regulating miR-741-3p and targeting Radil gene.

Key words: Schwann cell, electrical stimulation, miR-741-3p, Radil, cell migration, nerve regeneration

CLC Number:

Liu Qing, Gao Bo, Yang Xiao, Jiang Yu, Wang Pei. Electrical stimulation induces miR-741-3p to regulate Radil and promote Schwann cell migration[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(19): 4038-4043.

Figures/Tables 1

2.1 电刺激可抑制神经残端 miR-741-3p的表达图1中蓝色代表细胞核，绿色代表miR-741-3p，荧光强度越高代表表达水平越高，用Image J软件定量分析荧光强度得出电刺激组绿色的平均荧光强度为13±5，对照组绿色的平均荧光强度为34±11，差异有显著性意义(P < 0.05)，提示电刺激可抑制神经残端 miR-741-3p的表达。 2.2 miR-741-3p靶基因预测结果运用TargetScan、miRWalk、miRDB 3个数据库进行miR-741-3p的靶基因预测，最后取3个数据库的交集作为靶基因的最终预测结果，共69个基因可能为miR-741-3p的靶基因，Radil是预测的靶基因之一。另外，还查询了miR-741-3p和Radil的基因序列，发现miR-741-3p和Radil的3’端具有互补碱基序列，提示miR-741-3p与Radil可能存在靶向关系，见图2。 2.3 miR-741-3p在体外抑制施万细胞的迁移并可负向调节Radil的表达 RT-PCR检测结果显示miR-741-3p模拟物组miR-741-3p的表达明显上调，miR-741-3p抑制物组miR-741-3p的表达显著下调，与相应对照组相比差异有显著性意义(P < 0.001)。RT-PCR及Transwell实验结果显示与miR-741-3p抑制物对照组相比，miR-741-3p抑制物组施万细胞的迁移能力增加，同时Radil的表达水平升高，差异有显著性意义(P < 0.01)；与miR-741-3p模拟物对照组相比，miR-741-3p模拟物组施万细胞的迁移能力降低，同时Radil的表达降低，差异有显著性意义(P < 0.001)，见图3。 2.4 Radil在施万细胞迁移中起到重要的作用 RT-PCR检测结果显示Radil siRNA能够下调施万细胞Radil的表达(P < 0.001)。Transwell实验显示，与siRNA对照组相比，Radil siRNA组施万细胞的迁移能力明显减弱，siRNA对照组与Radil siRNA组施万细胞的迁移数分别为137±33，95±12，差异有显著性意义(P < 0.01)，见图4。 2.5 miR-741-3p抑制物促进施万细胞迁移的作用被Radil siRNA逆转如图5所示，miR-741-3p抑制物+Radil siRNA组和miR-741-3p抑制物+siRNA对照组施万细胞迁移数分别为87±19，133±25，差异有显著性意义(P < 0.01)。结果表明，miR-741-3p抑制物促进施万细胞迁移的作用可被Radil siRNA逆转，进一步确定miR-741-3p抑制物促进施万细胞的迁移依赖于其促进Radil的表达。"

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