A型有髓纤维DRG神经元内Cav3.2通道N-糖基化介导糖尿病触诱发痛的研究

doi:10.3969/j.issn.1674-8115.2025.11.005

上海交通大学学报（医学版） ›› 2025, Vol. 45 ›› Issue (11): 1458-1465.doi: 10.3969/j.issn.1674-8115.2025.11.005

• 论著 · 基础研究 • 上一篇

A型有髓纤维DRG神经元内Cav3.2通道N-糖基化介导糖尿病触诱发痛的研究

廖陈龙, 李烁, 许新, 张越, 张文川()

上海交通大学医学院附属第九人民医院神经外科，上海 200011

收稿日期:2025-06-02 接受日期:2025-07-17 出版日期:2025-11-28 发布日期:2025-12-03
通讯作者: 张文川，教授，主任医师，博士；电子信箱：zhangwench88@sjtu.edu.cn。
基金资助:
国家自然科学基金(81801219);国家自然科学基金(82271389);上海市“科技创新行动计划”医学创新研究专项(21Y11906300);上海市自然科学基金(21ZR1438100)

N-glycosylation of Cav3.2 channel in DRG neurons of myelinated A-fiber contributes to diabetic mechanical allodynia

LIAO Chenlong, LI Shuo, XU Xin, ZHANG Yue, ZHANG Wenchuan()

Department of Neurosurgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China

Received:2025-06-02 Accepted:2025-07-17 Online:2025-11-28 Published:2025-12-03
Contact: ZHANG Wenchuan, E-mail: zhangwench88@sjtu.edu.cn.
Supported by:
National Natural Science Foundation of China(81801219);Medical Innovation Research Project of Science and Technology Commission of Shanghai Municipality(21Y11906300);Natural Science Foundation of Shanghai(21ZR1438100)

摘要/Abstract

摘要：

目的·探索A型有髓纤维相关背根神经节（dorsal root ganglion，DRG）感觉神经元内Cav3.2通道N-糖基化与糖尿病触诱发痛（mechanical allodynia，MA）的关系。方法·通过单次腹腔注射链脲佐菌素建立1型糖尿病大鼠模型，每周进行疼痛行为学评估，包括MA和热痛觉过敏（thermal hyperalgesia，TH）评估。于第3周将筛选出的TH（+）糖尿病大鼠，进一步分为MA（+）与MA（-）2个亚组。采用树脂毒素（resiniferatoxin，RTX）选择性阻断无髓纤维后，通过先后使用神经氨酸酶（neuraminidase，NEU）和T型钙通道选择性阻断剂（T-type calcium channel inhibitor，TTA-P2），观察N-糖基化对A型有髓纤维相关DRG神经元Cav3.2通道表达和疼痛行为的影响。通过分析去糖基化后Cav3.2的N-末端片段，定量比较MA（+）与MA（-）糖尿病大鼠Cav3.2 N-糖基化的程度。结果·MA（+）组A型有髓纤维DRG神经元中的Cav3.2蛋白表达水平显著高于MA（-）组（P<0.001）。NEU诱导的去糖基化降低了各组Cav3.2的表达，但MA（+）组仍较MA（-）组糖尿病大鼠高（P<0.05）。RTX仅消除TH，而NEU和TTA-P2可同时逆转TH和MA。去糖基化处理后，游离的N端片段水平在MA（+）组中显著高于MA（-）组（P<0.001）。结论·A型有髓纤维相关DRG神经元中Cav3.2通道的N-糖基化介导了糖尿病触诱发痛的发生和发展。

关键词: 糖尿病周围神经病变, Cav3.2通道, 触诱发痛, 背根神经节, 糖基化

Abstract:

Objective ·To investigate the association of Cav3.2 channel N-glycosylation in myelinated A-fiber dorsal root ganglion (DRG) neurons with diabetic mechanical allodynia (MA). Methods ·Diabetic rat models were induced by a single intraperitoneal injection of streptozotocin. Pain behavior assessments, including evaluations of MA and thermal hyperalgesia (TH), were conducted on a weekly basis. Diabetic rats exhibiting TH were subsequently categorized into two subcategories based on the onset of mechanical allodynia by the third week. The impact of N-glycosylation on both the expression of Cav3.2 channels in DRG neurons and pain behaviors was investigated through sequential administration of neuraminidase (NEU) and TTA-P2, a selective blocker of T-type calcium channels, after C-unmyelinated fiber blockage by the application of resiniferatoxin (RTX). The extent of Cav3.2 N-glycosylation was quantitatively compared between diabetic rats with and without MA by analyzing the N-terminal fragment of Cav3.2 following de-glycosylation. Results ·Higher levels of Cav3.2 expression in myelinated A-fiber DRG neurons were observed in diabetic rats with MA compared to those without MA (P<0.001). NEU-induced de-glycosylation led to a reduction in Cav3.2 expression across all groups, with diabetic rats with MA maintaining higher levels of the Cav3.2 expression than those without MA (P<0.05). It was observed that RTX eliminated only TH and not MA, whereas NEU had a restorative effect on both TH and MA, similar to TTA-P2. Diabetic rats with MA exhibited increased expression of N-terminal fragments compared to those without MA (P<0.001). Conclusion ·N-glycosylation of the Cav3.2 channel within DRG neurons of myelinated A-fiber is a contributing factor to diabetic MA.

Key words: diabetic peripheral neuropathy, Cav3.2 channel, mechanical allodynia, dorsal root ganglion, glycosylation

中图分类号:

R587.2

廖陈龙, 李烁, 许新, 张越, 张文川. A型有髓纤维DRG神经元内Cav3.2通道N-糖基化介导糖尿病触诱发痛的研究[J]. 上海交通大学学报（医学版）, 2025, 45(11): 1458-1465.

LIAO Chenlong, LI Shuo, XU Xin, ZHANG Yue, ZHANG Wenchuan. N-glycosylation of Cav3.2 channel in DRG neurons of myelinated A-fiber contributes to diabetic mechanical allodynia[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(11): 1458-1465.

图/表 4

图1 各组大鼠血糖、体质量和疼痛行为学变化Note： A. Changes in blood glucose levels. B. Changes in body weight. C. Comparison of PWT. D. Comparison of PWL. ①P<0.001, ②P=0.006, compared with the control group; ③P<0.001, compared with the control group and Diabetic MA (-) group; ④P<0.001, compared with the two diabetic groups; ⑤P<0.001.

Fig 1 Changes in blood glucose, body weight, and pain behavioral measurements among groups

图2 Cav3.2钙通道在DRG神经元中的表达Note： A. Cav3.2 expression was rarely detected in the control group, whereas distinct expression patterns were observed in Diabetic MA (+) and Diabetic MA (-) subgroups. B. Expression of Cav3.2 in NF-200+ DRG neurons after NEU-induced de-glycosylation. Cav3.2 expression remained low in the control group and a further reduction was observed in both diabetic subgroups. ①P<0.001, ②P=0.038, ③P=0.005, ④P=0.006.

Fig 2 Expression of Cav3.2 in DRG neurons

图3 NEU、RTX和TTA-P2对于糖尿病大鼠疼痛行为学的影响Note： A/B. Effect of NEU on PWL (A) and PWT (B) in rats receiving STZ/vehicle injections. ①P<0.001, ②P=0.006, ③P=0.004, ④P=0.003, compared with Diabetic MA (-) group. C. Development of TH in diabetic subgroups was eliminated by pretreatment of RTX. ①P<0.001, compared with the VEH+STZ group. D. While not affected by pretreatment with RTX, MA in diabetic rats was reversed by administration of either TTA-P2 or NEU. ①P<0.001, compared with the control rats.

Fig 3 Effect of NEU, RTX, and TTA-P2 on pain behaviors in diabetic rats

图4 Western blotting检测Cav3.2钙通道N端片段的表达水平Note： A. Representative bands. B. Quantificational results of N-terminal fragments of Cav3.2 in all groups. ①P<0.001.

Fig 4 Western blotting analysis of N-terminal fragments of Cav3.2

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A型有髓纤维DRG神经元内Cav3.2通道N-糖基化介导糖尿病触诱发痛的研究

N-glycosylation of Cav3.2 channel in DRG neurons of myelinated A-fiber contributes to diabetic mechanical allodynia

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