Study on the role of FGF13 in the repair of inferior alveolar nerve injury by regulating mitochondrial function

doi:10.12439/kqhm.1005-4979.2024.05.003

Abstract

Abstract: [Abstract] Objective: To explore the role and mechanism of fibroblast growth factor 13 (FGF13) in the repair of inferior alveolar nerve injury. Methods: To establish the model of inferior alveolar nerve injury in SD (Sprague-Dawley) rats. RNA from inferior alveolar tissue samples was collected 1, 3, 7 days after injury. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to detect the transcription expression levels of FGF13 gene at different time points after injury. Primary trigeminal nerve cells of neonatal SD mice were extracted and divided into experimental group and control group, transfected with FGF13 overexpressed plasmid and control plasmid, respectively. 3 days after successful transfection, cell RNA was extracted to detect the gene expression level of neurotrophin by RT-qPCR. Trigeminal nerve cells were stained with Neun and βⅢ-Tubulin nerve immunofluorescence, and the axon length of nerve cells was observed by laser scanning confocal microscope. ND7/23 nerve cells were divided into overexpression group (ND7/23-FGF13) and control group (ND7/23-vector), transfected with FGF13 overexpression lentivirus and control virus, respectively. Stable transmutation strains were screened out by purinomycin, and FGF13 protein immunofluorescence staining and JC-1 mitochondrial membrane potential fluorescence probe staining were performed. Extract cell RNA, then RT-qPCR was used to detect the expression of mitophagy related genes. Results: The expression level of FGF13 was significantly increased at 1 day after inferior alveolar nerve injury, decreased at 3 days after injury, and decreased to an equivalent level to the control group at 7 days after injury. Compared with the control group, the expression of FGF13, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and other factors increased in the experimental group. In the lentivirus overexpression group, FGF13 protein was more densely distributed in the nucleus, mitochondrial membrane potential was significantly increased, and the expression of mitochondrial autophagy related genes was increased. Conclusion: After inferior alveolar nerve injury, the expression level of FGF13 is transiently increased, which may have potential significance for the repair process of inferior alveolar nerve. Overexpression of FGF13 in trigeminal nerve cells can promote axonal elongation of nerve cells. The underlying mechanism may be related to the regulation of mitochondrial function and the promotion of mitochondrial homeostasis.

Key words: inferior alveolar nerve injuries, fibroblast growth factor 13, mitochondria

摘要：

目的：探究成纤维细胞生长因子 13（fibroblast growth factor 13，FGF13）在下牙槽神经损伤修复中的作用及机制。方法：建立 SD（Sprague-Dawley）大鼠下牙槽神经损伤模型，在损伤后 1、3、7 d 时收取下牙槽神经组织样本 RNA，实时定量聚合酶链反应（real-time quantitative polymerase chain reaction，RT-qPCR) 检测损伤后不同时间 FGF13 基因的表达水平；提取新生 SD 大鼠的原代三叉神经细胞，并将其分为实验组和对照组，分别转染 FGF13 过表达质粒和对照质粒，转染成功后 3 d 提取细胞 RNA，利用 RT-qPCR 检测神经营养因子的基因表达水平，并对三叉神经细胞进行 Neun 和βⅢ-Tubulin 免疫荧光染色，使用激光扫描共聚焦显微镜摄片，观测神经细胞的轴突长度；将 ND7/23 神经细胞分为过表达组（ND7/23-FGF13）和对照组（ND7/23-vector），分别转染 FGF13 过表达慢病毒和对照病毒，嘌呤霉素筛出稳转株，进行 FGF13 蛋白免疫荧光染色和 JC-1 线粒体膜电位荧光探针染色，提取细胞 RNA，利用 RT-qPCR 检测线粒体自噬相关基因的表达。结果：下牙槽神经损伤后 1 d，该组织的 FGF13 表达量显著升高，损伤后 3 d 表达量降低，损伤后 7 d表达量降至对照组水平；实验组与对照组相比，其 FGF13、神经生长因子（nerve growth factor，NGF）、脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）等表达量升高，实验组细胞轴突长度大于对照组；慢病毒过表达组与对照组相比，FGF13 蛋白在细胞核中分布更密集，线粒体膜电位明显升高，线粒体自噬相关基因表达量升高。结论：下牙槽神经损伤后，FGF13 表达量出现一过性升高，其对下牙槽神经的修复过程可能具有潜在意义；在三叉神经细胞中过表达 FGF13 可以促进神经细胞的轴突伸长，其内在机制可能与调节线粒体功能、促进线粒体稳态有关。

关键词:

下牙槽神经损伤, 成纤维细胞生长因子13, 线粒体

CLC Number:

R782

JIAO Yi, SUN Xinrong, LIU Weicai.

Study on the role of FGF13 in the repair of inferior alveolar nerve injury by regulating mitochondrial function

[J]. 《Journal of Oral and Maxillofacial Surgery》, 2024, 34(5): 342-349.

焦祎, 孙欣荣, 刘伟才.

FGF13通过调节线粒体功能对下牙槽神经损伤修复的作用研究

[J]. 《口腔颌面外科杂志》, 2024, 34(5): 342-349.

/ / Recommend / Download Citations

URL: https://journal06.magtech.org.cn/Jweb_joms/EN/10.12439/kqhm.1005-4979.2024.05.003

https://journal06.magtech.org.cn/Jweb_joms/EN/Y2024/V34/I5/342

References

[1] Li JW, Wang QQ, Wang HL, et al. Lentivirus mediating FGF13 enhances axon regeneration after spinal cord injury by stabilizing microtubule and improving mitochondrial function[J]. J Neurotrauma, 2018, 35(3): 548-559.
[2] 王艳颖, 宫苹, 张健. 种植体周围神经再生的研究进展 [J]. 现代口腔医学杂志, 2019, 33(2): 107-110.
[3] Arthur-Farraj P, Coleman MP. Lessons from injury: How nerve injury studies reveal basic biological mechanisms and therapeutic opportunities for peripheral nerve diseases[J]. Neurotherapeutics, 2021, 18(4): 2200-2221.
[4] Wan QQ, Qin WP, Ma YX, et al. Crosstalk between bone and nerves within bone[J]. Adv Sci, 2021, 8(7): 2003390.
[5] Zang YX, Chaudhari K, Bashaw GJ. New insights into the molecular mechanisms of axon guidance receptor regulation and signaling[J]. Curr Top Dev Biol, 2021, 142: 147-196.
[6] 刘姗姗, 林玮, 周剑辉, 等. 成纤维细胞生长因子及其在代谢性疾病中的作用与临床应用研究进展 [J]. 药学进展, 2019, 43(1): 19-28.
[7] Zhang X, Bao L, Yang L, et al. Roles of intracellular fibroblast growth factors in neural development and functions[J]. Sci China Life Sci, 2012, 55(12): 1038-1044.
[8] Yang L, Dong F, Yang Q, et al. FGF13 selectively regulates heat nociception by interacting with Nav1.7[J]. Neuron, 2017, 93(4): 806-821.e9.
[9] Wu QF, Yang L, Li S, et al. Fibroblast growth factor 13 is a microtubule-stabilizing protein regulating neuronal polarization and migration[J]. Cell, 2012, 149(7): 1549-1564.
[10] Li R, Tao XT, Huang MH, et al. Fibroblast growth factor 13 facilitates peripheral nerve regeneration through maintaining microtubule stability[J]. Oxid Med Cell Longev, 2021, 2021: 5481228.
[11] Li RM, Xiao L, Zhang T, et al. Overexpression of fibroblast growth factor 13 ameliorates amyloid-β-induced neuronal damage[J]. Neural Regen Res, 2023, 18(6): 1347-1353.
[12] Cheng XT, Sheng ZH. Developmental regulation of microtubule-based trafficking and anchoring of axonal mitochondria in health and diseases[J]. Dev Neurobiol, 2021, 81(3): 284-299.
[13] Wang BY, Huang MH, Shang DH, et al. Mitochondrial behavior in axon degeneration and regeneration[J]. Front Aging Neurosci, 2021, 13: 650038.
[14] Mishra E, Thakur MK. Mitophagy: A promising therapeutic target for neuroprotection during ageing and age-related diseases[J]. Br J Pharmacol, 2023, 180(12): 1542-1561.
[15] Ravera S, Colombo E, Pasquale C, et al. Mitochondrial bioenergetic, photobiomodulation and trigeminal branches nerve damage, what's the connection? A review[J]. Int J Mol Sci, 2021, 22(9): 4347.

Please choose a citation manager

Content to export