减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响

doi:10.3969/j.issn.1006-9771.2021.02.002

《中国康复理论与实践》 ›› 2021, Vol. 27 ›› Issue (2): 131-136.doi: 10.3969/j.issn.1006-9771.2021.02.002

减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响

李向哲¹,丁洁²,王庆华³,董传明⁴,王彤⁵,吴勤峰¹()

^1.南京医科大学附属苏州科技城医院，江苏苏州市 215153
^2.浙江大学医学院附属邵逸夫医院，浙江杭州市 310016
^3.南通大学实验动物中心，江苏南通市 226001
^4.南通大学医学院，江苏南通市 226001
^5.南京医科大学第一附属医院康复医学中心，江苏南京市 210029

收稿日期:2019-12-03 修回日期:2020-12-28 出版日期:2021-02-25 发布日期:2021-02-26
通讯作者: 吴勤峰 E-mail:wuqinfeng0911@163.com
作者简介:李向哲(1988-)，男，汉族，河南汝阳县人，硕士，医师，主要研究方向：神经损伤的康复研究|吴勤峰(1976-)，男，汉族，黑龙江哈尔滨市人，博士，副教授，副主任医师，主要研究方向：神经损伤及肌骨疼痛的康复与治疗。
基金资助:
苏州市"科教兴卫"青年科技项目(KJXW2018082);苏州市科技局民生科技项目(SYS201785);国家自然科学基金面上项目(81672258)

Effects of Body Weight-supported Treadmill Training on Neuropathic Pain and Expression of Glutamate Decarboxylase (GAD)-65/67 in Spinal Dorsal Horn of Rats with Spinal Cord Injury

Xiang-zhe LI¹,Jie DING²,Qing-hua WANG³,Chuan-ming DONG⁴,Tong WANG⁵,Qin-feng WU¹()

^1.the Affiliated Suzhou Science and Technology Town Hospital of Nanjing Medical University, Suzhou, Jiangsu 215153, China
^2.Sir Run Run Shaw Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, China
^3.Experimental Animal Center of Nantong University, Nantong, Jiangsu 226001, China
^4.Medical College of Nantong University, Nantong, Jiangsu 226001, China
^5.Rehabilitation Medical Center, the First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China

Received:2019-12-03 Revised:2020-12-28 Published:2021-02-25 Online:2021-02-26
Contact: Qin-feng WU E-mail:wuqinfeng0911@163.com
Supported by:
Suzhou "Kejiaoxingwei" Youth Science and Technology Project(KJXW2018082);People's Livelihood Science and Technology Project of Suzhou Science and Technology Bureau(SYS201785);National Natural Science Foundation of China (General)(81672258)

摘要/Abstract

摘要： 目的

探讨减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角内谷氨酸脱羧酶-65/67 (GAD-65/67)表达的影响。

方法

将24只Sprague-Dawley大鼠随机分为假手术组(Sham组)、脊髓损伤-不运动组(SCI-Sed组)和脊髓损伤-运动组(SCI-Ex组)，每组8只。采用Allen法制作T₁₀不完全性脊髓损伤模型。在脊髓损伤后7 d对SCI-Ex组进行减重平板训练。于脊髓损伤前，脊髓损伤后7 d、14 d、21 d、28 d和35 d，分别采用von Frey细丝和热刺激痛觉测试仪进行机械性痛阈和热痛阈评估。痛阈评估结束后，采用Western blotting和免疫组化染色检测所有大鼠腰髓GAD-65和GAD-67。

结果

脊髓损伤后21 d、28 d和35 d，SCI-Ex组机械性痛阈和热痛阈均明显高于SCI-Sed组(P < 0.01)。与Sham组相比，SCI-Sed组和SCI-Ex组GAD-65和GAD-67表达减少(P < 0.05)；与SCI-Sed组相比，SCI-Ex组GAD-65和GAD-67表达量升高(P < 0.05)。

结论

减重平板训练可增加不完全性脊髓损伤大鼠损伤远端脊髓后角GAD-65/67的合成，并改善大鼠后肢痛阈。

关键词: 脊髓损伤, 减重平板训练, 神经病理性疼痛, γ-氨基丁酸, 谷氨酸脱羧酶-65/67, 大鼠

Abstract: Objective

To investigate the effect of body weight-supported treadmill training on neuropathic pain and expression of glutamate decarboxylase-65/67 (GAD-65/67) in spinal dorsal horn of rats with spinal cord injury (SCI).

Methods

A total of 24 Sprague-Dawley rats were randomly divided into sham group, SCI-sedentary (SCI-Sed) group and SCI-Exercise (SCI-Ex) group, with eight rats in each group. Allen's method was used to make T₁₀ incomplete SCI model. Seven days after SCI, SCI-Ex group was given body weight-supported treadmill training. Before SCI, and seven days, 14 days, 21 days, 28 days and 35 days after SCI, the von Frey filaments and thermal stimulation pain tester were used to evaluate the mechanical and thermal pain thresholds. Then, Western blotting and immunohistochemical analysis were performed on the spinal cord of all rats to detect the expression of GAD-65 and GAD-67.

Results

The mechanical and thermal pain thresholds were higher in SCI-Ex group than in SCI-Sed group 21 days, 28 days and 35 days after SCI (P < 0.01). Compared with the sham group, the expression of GAD-65 and GAD-67 decreased in SCI and SCI-Ex groups (P < 0.05), and increased in SCI-Ex group compared with that of SCI-Sed group (P < 0.05).

Conclusion

Body weight-supported treadmill training could increase the synthesis of GAD-65/67 in the distal spinal cord dorsal horn of incomplete SCI rats, and improve the pain thresholds of hind limbs in rats with SCI.

Key words: spinal cord injury, body weight-supported treadmill training, neuropathic pain, γ-aminobutyric acid, glutamate decarboxylase-65/67, rats

中图分类号:

R651.2

李向哲,丁洁,王庆华,董传明,王彤,吴勤峰. 减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 131-136.

Xiang-zhe LI,Jie DING,Qing-hua WANG,Chuan-ming DONG,Tong WANG,Qin-feng WU. Effects of Body Weight-supported Treadmill Training on Neuropathic Pain and Expression of Glutamate Decarboxylase (GAD)-65/67 in Spinal Dorsal Horn of Rats with Spinal Cord Injury[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(2): 131-136.

图/表 6

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参考文献 29

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组别	n	术前	7 d	14 d	21 d	28 d	35 d
Sham组	8	6.50±1.41	6.50±1.41	6.75±1.49	6.25±1.28	6.75±1.49	6.50±0.93
SCI-Sed组	8	6.25±1.67	2.05±1.91^a	0.70±0.26^a	0.50±0.11^a	0.63±0.25^a	0.70±0.26^a
SCI-Ex组	8	6.25±1.67	1.95±1.97^a	1.93±1.72^a	2.05±0.87^a,b	2.93±1.17^a,c	3.50±1.41^a,c
F值		0.066	16.952	46.941	87.881	63.191	69.016
P值		0.936	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	n	术前	7 d	14 d	21 d	28 d	35 d
Sham组	8	15.45±3.55	15.31±5.50	14.58±2.87	15.55±2.35	15.74±2.77	16.92±1.63
SCI-Sed组	8	15.72±4.47	3.55±2.46^a	4.05±2.98^a	3.47±2.20^a	4.82±1.66^a	4.45±2.37^a
SCI-Ex组	8	15.48±2.97	4.47±2.88^a	5.99±5.36^a	9.38±3.25^a,b	10.41±3.08^a,b	10.86±2.62^a,c
F值		0.012	23.036	16.429	41.902	36.036	61.724
P值		0.988	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

组别	n	GAD-65	GAD-67
Sham组	4	1.00±0.18	1.00±0.10
SCI-Sed组	4	0.41±0.08^b	0.43±0.07^c
SCI-Ex组	4	0.61±0.09^a,d	0.72±0.06^b,d
F值		22.900	51.693
P值		< 0.001	< 0.001

组别	n	GAD-65	GAD-67
Sham组	4	1.00±0.19	1.00±0.23
SCI-Sed组	4	0.46±0.18^b	0.27±0.18^b
SCI-Ex组	4	0.76±0.14^a,c	0.56±0.15^b,c
F值		30.351	45.162
P值		< 0.001	< 0.001

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减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响

Effects of Body Weight-supported Treadmill Training on Neuropathic Pain and Expression of Glutamate Decarboxylase (GAD)-65/67 in Spinal Dorsal Horn of Rats with Spinal Cord Injury

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