单侧前牙反𬌗对三叉神经感觉主核背内侧部及丘脑腹后内侧核VGLUT表达水平影响的研究

doi:10.3969/j.issn.1005-4979.2020.06.004

《口腔颌面外科杂志》 ›› 2020, Vol. 30 ›› Issue (6): 362-368. doi: 10.3969/j.issn.1005-4979.2020.06.004

单侧前牙反𬌗对三叉神经感觉主核背内侧部及丘脑腹后内侧核VGLUT表达水平影响的研究

时铭鸿¹(), 刘鑫²^,³, 谢勉姣², 王冬梅¹, 李金莲⁴, 王美青¹^,²()

¹ 新乡医学院第三附属医院口腔科，河南新乡 453000
² 第四军医大学口腔医学院口腔解剖生理学教研室，陕西西安 710032
³ 中国人民解放军第960医院口腔科，山东济南 250031
⁴ 第四军医大学人体解剖与组织胚胎学教研室，陕西西安 710032

收稿日期:2020-02-05 修回日期:2020-06-06 出版日期:2020-12-28 发布日期:2024-03-20
通讯作者: 王美青，教授. E-mail: mqwang@fmmu.edu.cn
作者简介:
时铭鸿（1993—），女，山东人，硕士. E-mail: s1144632@163.com
基金资助:
国家自然科学基金项目(81870866)

Effect of Unilateral Anterior Crossbite on VGLUT Expression in the Dorsomedial Part of the Trigeminal Nucleus and Ventral Posteromedial Thalamic Nucleus

SHI Minghong¹(), LIU Xin²^,³, XIE Mianjiao², WANG Dongmei¹, LI Jinlian⁴, WANG Meiqing¹^,²()

¹ Department of Stomatology, The Third Affiliated Hospital, Xinxiang Medical College, Xinxiang 453000, Henan Province
² Department of Oral Anatomy and Physiology, School of Stomatology,The Fourth Military Medical University, Xi′an 710032, Shaanxi Province
³ Department of Stomatology, the 960th Hospital of People′s Liberation Army, Jinan 250031, Shandong Province
⁴ Department of Anatomy, Histology and Embryology, The Fourth Military Medical University, Xi′an 710032, Shaanxi Province, China

Received:2020-02-05 Revised:2020-06-06 Online:2020-12-28 Published:2024-03-20

摘要/Abstract

摘要：

目的：异常咬合信息可以通过牙周本体感受器、三叉神经系向高级中枢传递，三叉神经感觉主核背内侧部（dorsomedial part of the trigeminal nucleus, Vpdm）-丘脑腹后内侧核（ventral posteromedial thalamic nucleus, VPM）是重要的三叉神经系上传通路，Vpdm是否会受到异常咬合影响，是否继续上传至VPM值得深入研究。本文通过检测单侧前牙反𬌗（unilateral anterior crossbite, UAC）大鼠，Vpdm及VPM中兴奋性神经传导标志物—囊泡膜谷氨酸转运体1,2 （vesicular glutamate transporter1,2, VGLUT1,2）的表达变化，探讨该问题。方法：将雌性大鼠42只分为UAC组和对照组，造模后2、4和8周取材，以免疫荧光、Western blot及实时定量PCR技术，检测目标神经核团中VGLUT1,2的表达差异。结果：UAC组Vpdm内VGLUT1蛋白和 mRNA的表达在各个时间点均明显高于对照组，而VGLUT2 mRNA的表达则无显著性差异。在VPM内，VGLUT1,2的蛋白水平和mRNA水平两组间均无显著差异。结论：UAC促进了Vpdm的VGLUT1表达，但对VPM的VGLUT1,2表达水平没有显著影响。

关键词: 咬合, 三叉神经感觉主核背内侧部, 丘脑腹后内侧核, 囊泡膜谷氨酸转运体, 三叉神经传导通路

Abstract:

Objective: Periodontal proprioceptional message derived from abnormal occlusion is transmitted via projections from mesencephalic trigeminal nucleus to dorsomedial part of the trigeminal nucleus(Vpdm) and then to ventral posteromedial thalamic nucleus(VPM). To elucidate whether abnormal occlusion will send impact on Vpdm and then on VPM, the expression levels of vesicular glutamate transporter1,2(VGLUT1,2), markers of excitatory nucleus, were detected in Vpdm and VPM of the unilateral anterior crossbite(UAC) treated rats. Methods: The female rats were divided into UAC and control group, and were killed at 2, 4 and 8 week respectively. The expression levels of VGLUT1,2 were detected through immunofluorescence, Western blot and real-time PCR assays. Results: In UAC group, the VGLUT1 protein and mRNA expression levels in Vpdm were increased at all the three time points, while that of VGLUT2 showed no significant differences from the age-matched controls. No such a difference was noticed in the expression levels of VGLUT1,2 in VPM at both protein and mRNA levels. Conclusions: UAC promoted VGLUT1 expression in Vpdm, but showed no significant impact on the expression of VGLUT1,2 in VPM.

Key words: occlusion, dorsomedial part of the trigeminal nucleus(Vpdm), ventral posteromedial thalamic nucleus (VPM), vesicular glutamate transporter (VGULT), trigeminal pathway

时铭鸿, 刘鑫, 谢勉姣, 王冬梅, 李金莲, 王美青. 单侧前牙反𬌗对三叉神经感觉主核背内侧部及丘脑腹后内侧核VGLUT表达水平影响的研究[J]. 《口腔颌面外科杂志》, 2020, 30(6): 362-368.

SHI Minghong, LIU Xin, XIE Mianjiao, WANG Dongmei, LI Jinlian, WANG Meiqing. Effect of Unilateral Anterior Crossbite on VGLUT Expression in the Dorsomedial Part of the Trigeminal Nucleus and Ventral Posteromedial Thalamic Nucleus[J]. 《Journal of Oral and Maxillofacial Surgery》, 2020, 30(6): 362-368.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2020/V30/I6/362

图/表 10

图1 下牙槽神经注射CTb动物的Vpdm处CTb、VGLUT1和NeuN三标情况注：A-D. 对照组和实验组CTb，VGLUT1和NeuN免疫荧光染色；C、D. 分别为A、B图中方框部位的放大成像。

Figure 1 Triple-immunofluorescence histochemistry for cholera toxin B subunit (CTb), vesicular glutamate transporter 1 (VGLUT1) and NeuN in Vpdm from animals injected with CTb in the inferior alveolar nerve

图2 UAC实验组和对照组Vpdm染色结果比较注：A. 实验组和对照组阳性细胞数的比较；B. 实验组和对照组阳性细胞百分比的比较。与对照组比较，* P<0.05，** P<0.01，*** P<0.001。

Figure 2 Comparison of the Vpdm staining between UAC and control group

图3 Vpdm定点注射BDA（A）后对侧VPM处BDA的表达情况（B）注：A. BDA注射于Vpdm；B. BDA在VPM的纤维投射。

Figure 3 Injection of biotinylated dextran BDA at Vpdm (A) and the BDA positive fibers at the contra-lateral VPM (B)

图4 UAC 实验组和对照组之间Vpdm处注射BDA后对侧VPM处BDA、VGLUT1和NeuN三标图比较注：A-D. 对照组和实验组BDA，VGLUT1和NeuN免疫荧光染色；C、D. 分别为A、B图中方框部位的放大成像。

Figure 4 Triple-immunofluorescence histochemistry for biotinylated dextran (BDA), vesicular glutamate transporter 1 (VGLUT1) and neuronal nuclei (NeuN) at contralateral side VPM, which were compared between the UAC group and the control group after BDA injected at Vpdm

图5 UAC实验组和对照组VPM染色结果比较注：A. 实验组和对照组阳性细胞数的比较；B. 实验组和对照组阳性细胞百分比的比较。

Figure 5 Comparison of the VPM staining between UAC and control groups

图6 2、4和8周时间点对照组和UAC组的Vpdm中实时定量PCR检测的VGLUT1，2的mRNA表达水平比较注：A. 实时定量PCR检测实验组和对照组中VGLUT1 mRNA表达水平；B. VGLUT2 mRNA表达水平。与对照组比较，* P<0.05，** P<0.01，*** P<0.001。

Figure 6 Comparison of the mRNA expression level of VGLUT1,2 in Vpdm between control and UAC group at 2, 4, and 8 weeks detected by real-time PCR

图7 2、4和8周时间点对照组和UAC组的VPM中实时定量PCR检测的VGLUT1，2 mRNA表达水平比较注：A. 实验组和对照组VPM中VGLUT1 mRNA表达水平；B. VGLUT2 mRNA表达水平。

Figure 7 Comparison of the mRNA expression level of VGLUT1,2 in VPM between control and UAC group at 2, 4, and 8 weeks detected by real-time PCR

图8 2、4和8周时间点对照组和UAC组的Vpdm中Western blot 检测的VGLUT1蛋白表达水平比较注：A. 用Western blot检测VGLUT1蛋白表达情况；B. Western blot条带分析。与对照组相比，**P<0.01，*** P<0.001。

Figure 8 Comparison of the expression level of VGLUT1,2 protein in Vpdm between control and UAC group at 2, 4, and 8 weeks detected by Western blot assay

图9 2、4和8周时间点对照组和UAC组的VPM中Western blot 检测的VGLUT1，2蛋白表达水平比较注：A. 应用Western blot检测VGLUT1蛋白表达情况；B. 应用Western blot检测VGLUT2蛋白表达情况；C. Western blot检测VGLUT1条带分析；D. Western blot检测VGLUT2条带分析。

Figure 9 Comparison of the expression level of VGLUT1,2 protein in VPM between control and UAC group at 2, 4, and 8 weeks detected by Western blot assay

图10 本文针对Vme-Vpdm-VPM神经传导通路研究的结论示意图注：UAC通过牙周本体感受器兴奋三叉神经中脑核（Vme），继而通过囊泡膜谷氨酸转运体1（VGLUT1）向三叉神经感觉主核背内侧部（Vpdm）发出兴奋性信号（本文结果），并通过VGLUT向第Ⅴ, Ⅶ, Ⅺ, Ⅻ对脑神经运动核发出兴奋性信号^[10-11]，而没有证据显示UAC 大鼠的Vpdm以VGLUT1,2向丘脑腹后内侧核（VPM）发出兴奋性信号。

Figure 10 Diagram for the obtaining from the present data focusing on Vme-Vpdm-VPM pathway

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单侧前牙反𬌗对三叉神经感觉主核背内侧部及丘脑腹后内侧核VGLUT表达水平影响的研究

Effect of Unilateral Anterior Crossbite on VGLUT Expression in the Dorsomedial Part of the Trigeminal Nucleus and Ventral Posteromedial Thalamic Nucleus

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