南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (3): 571-577.doi: 10.12122/j.issn.1673-4254.2024.03.19

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miR-132-3p/CAMTA1对I-125粒子处理的面神经损伤大鼠施万细胞的调控作用

朱 瑾，欧阳欣，刘 屿，钱叶梅，夏 斌，施延安，俞力夫   

  1. 昆明医科大学附属口腔医院颌面外科，云南 昆明 650106；云南省第一人民医院口腔医学中心，云南 昆明 6500321
  • 出版日期:2024-03-20 发布日期:2024-04-03

MiR-132-3p negatively regulates CAMTA1 to promote Schwann cell proliferation and migration and alleviates I-125 seeds-induced exacerbation of facial nerve injury in rats

ZHU Jin, OUYANG Xin, LIU Yu, QIAN Yemei, XIA Bin, SHI Yanan, YU Lifu   

  1. Department of Maxillofacial Surgery, Stomatological Hospital of Kunming Medical University, Kunming 650106, China; Stomatology Center, First People's Hospital of Yunnan Province, Kunming 650032, China
  • Online:2024-03-20 Published:2024-04-03

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摘要： 目的 探讨miR-132-3p通过钙调素结合转录因子1（CAMTA1）对I-125粒子处理的面神经损伤大鼠（FNI）中施万细胞的调控作用。方法 用I-125粒子辐射大鼠施万细胞，并在细胞中转染miR-132-3p mimic、miR-132-3p inhibitor以及sh-CAMTA1。免疫荧光检测S100B和β-Tubulin Ⅲ荧光强度。RT-qPCR检测miR-132-3p的表达，Western blot检测CAMTA1蛋白表达。EdU染色评估细胞增殖，Transwell检测细胞迁移。同时，构建FNI大鼠模型并在大鼠面部植入I-125粒子，HE、LFB染色以及IF染色评估大鼠面神经组织的病理损伤。StarBase v2.0数据库和双荧光素酶报告实验验证miR-132-3p和CAMTA1的靶向关系。结果 S100B和β-Tubulin Ⅲ在大鼠施万细胞中显著表达。I-125粒子辐射组中miR-132-3p表达降低（P<0.001），抑制细胞的增殖（P<0.001）和迁移（P<0.001）。过表达miR-132-3p或敲降CAMTA1显著促进施万细胞的增殖（P<0.001）和迁移（P<0.05），敲降miR-132-3p则具有相反的作用。机制研究显示，miR-132-3p靶向负调控CAMTA1。体内实验结果显示，过表达miR-132-3p通过抑制CAMTA1的蛋白表达，减弱I-125粒子对FNI大鼠面神经损伤的促进作用。结论 过表达miR-132-3p抑制CAMTA1的表达，促进施万细胞的增殖和迁移，抑制I-125对FNI大鼠面神经损伤的促进作用。

关键词: 面神经损伤；miR-132-3p；施万细胞；钙调素结合转录因子1；增殖；迁移

Abstract: Objective To investigate the regulatory effect of miR-132-3p on calmodulin-binding transcription activator 1 (CAMTA1) and Schwann cell activity in rats with facial nerve injury (FNI) treated with I-125 seeds. Methods Rat Schwann cells were irradiated with I-125 seeds and transfected with miR-132-3p mimic, miR-132-3p inhibitor or sh-CAMTA1. The expressions of S100B and β-tubulin III in the cells were detected with immunofluorescence assay, and the expressions of miR-132-3p and CAMTA1 protein were determined using RT-qPCR and Western blotting, respectively. EdU staining and Transwell assay were used to evaluate the changes in cell proliferation and migration ability. In a rat model of FNI, I-125 seeds were implanted into the facial tissues near the facial nerve 2 weeks before modeling, and miR-132-3p mimic was injected subcutaneously in the face after modeling. The pathologies of the facial nerve was assessed by HE, LFB and immunofluorescence staining. The targeting relationship between miR-132-3p and CAMTA1 was verified using StarBase v2.0 database and dual-luciferase reporter assay. Results Rat Schwann cells showed high expressions of S100B and β- tubulin III. I-125 seeds radiation significantly decreased miR-132-3p expression and repressed proliferation and migration of the cells (P<0.001). Overexpression of miR-132-3p or CAMTA1 knockdown obviously enhanced proliferation and migration of the Schwann cells, while miR-132-3p knockdown produced the opposite effect. MiR-132-3p negatively regulated CAMTA1 expression. In the rat models of FNI, miR-132-3p injection significantly inhibited CAMTA1 expression and attenuated I-125 seeds-induced exacerbation of FNI. Conclusion Overexpression of miR-132-3p suppresses CAMTA1 expression and promotes Schwann cell proliferation and migration to alleviate I-125 seeds-induced exacerbation of FNI in rats.

Key words: facial nerve injury; miR-132-3p; schwann cells; calmodulin binding transcription activator 1; proliferation; migration