电针对紫杉醇诱导大鼠神经病理性疼痛的影响

doi:10.11958/20240609

天津医药 ›› 2024, Vol. 52 ›› Issue (11): 1141-1145.doi: 10.11958/20240609

电针对紫杉醇诱导大鼠神经病理性疼痛的影响

欧阳杰(), 赵海倩, 孔云, 牛钦, 陈莹, 思永玉^△()

昆明医科大学第二附属医院麻醉科（邮编650504）

收稿日期:2024-05-16 修回日期:2024-06-20 出版日期:2024-11-15 发布日期:2024-11-12
通讯作者: △E-mail：siyongyu@kmmu.edu.cn
作者简介:欧阳杰（1985），男，副主任医师，主要从事病理性疼痛干预方面研究。E-mail：oyangjie@kmmu.edu.cn
基金资助:
云南省科技厅昆明医科大学应用基础研究联合专项(202101AY070001-149);云南省科技厅昆明医科大学应用基础研究联合专项(202201AY070001-125)

The effect of electroacupuncture on paclitaxel-induced neuropathic pain in rats

OUYANG Jie(), ZHAO Haiqian, KONG Yun, NIU Qin, CHEN Ying, SI Yongyu<sup>△</sup>()

Department of Anesthesiology, the Second Affiliated Hospital of Kunming Medical University, Kunming 650504, China

Received:2024-05-16 Revised:2024-06-20 Published:2024-11-15 Online:2024-11-12
Contact: △E-mail：siyongyu@kmmu.edu.cn

欧阳杰, 赵海倩, 孔云, 牛钦, 陈莹, 思永玉. 电针对紫杉醇诱导大鼠神经病理性疼痛的影响[J]. 天津医药, 2024, 52(11): 1141-1145.

OUYANG Jie, ZHAO Haiqian, KONG Yun, NIU Qin, CHEN Ying, SI Yongyu. The effect of electroacupuncture on paclitaxel-induced neuropathic pain in rats[J]. Tianjin Medical Journal, 2024, 52(11): 1141-1145.

摘要/Abstract

摘要：

目的观察电针（EA）对紫杉醇诱导神经病理性疼痛大鼠脊髓背角NKCC1、KCC2表达和小胶质细胞活化的影响及其可能机制。方法将48只雄性SD大鼠随机分为溶媒组（Vehicle组）、紫杉醇组（PTX组）、紫杉醇+电针组（PTX+EA组）、紫杉醇+假电针组（PTX+Sham EA组），每组12只。采用腹腔注射PTX的方法建立PTX诱导的神经病理性疼痛大鼠动物模型，建模完成后，PTX+EA组给予“足三里”、“阳陵泉”电针刺激，连续7 d。于紫杉醇注射前2 d和注射后第1、3、5、7、14、21天进行机械撤足阈值和热缩足潜伏期痛行为学测试。利用免疫荧光染色技术和Western blot技术检测脊髓背角组织中钠钾氯联合转运蛋白1（NKCC1）、钾氯联合转运蛋白2（KCC2）和小胶质细胞标志物离子钙结合衔接分子1（Iba1）表达的变化。结果与Vehicle组比较，PTX组大鼠出现双后足机械和热痛觉过敏，脊髓背角组织中NKCC1表达升高和活化的小胶质细胞数增加。与PTX组比较，PTX+EA组大鼠在第14、21天的机械和热痛觉过敏得到显著改善，脊髓背角组织中NKCC1和Iba1表达量降低，4组间KCC2表达差异无统计学意义。结论电针可有效缓解紫杉醇诱导的神经病理性疼痛，其机制可能与抑制大鼠脊髓背角组织中NKCC1表达和小胶质细胞活化有关。

关键词: 电针, 紫杉醇, 神经痛, 脊髓背角, 小神经胶质细胞, 钠钾氯化物协同转运子, 神经病理性疼痛

Abstract:

Objective To observe the effect of electroacupuncture (EA) on the expression of NKCC1, KCC2 and activation of microglia in spinal dorsal horn of paclitaxel (PTX)-induced neuropathic pain rats and its possible mechanism. Methods Male SD rats were randomly divided into the vehicle group (vehicle), the PTX group, the PTX + EA group and the PTX + sham EA group, with 12 rats in each group. The rat model of PTX-induced neuropathic pain was established by intraperitoneal injection of PTX. After modeling, EA was applied to "Zusanli" and "Yanglingquan" for 7 days in the PTX + EA group. Paw withdrawal threshold and paw withdrawal latency were tested at 2 days before and 1, 3, 5, 7, 14 and 21 days after PTX injection. Immunofluorescence and Western blot assay were used to detect expression levels of sodium-potassium-chloride cotransporter 1 (NKCC1), potassium-chloride cotransporters 2 (KCC2) and microglia markers - ionized calcium binding adapter molecule 1 (Iba1) in spinal dorsal horn. Results Compared with the vehicle group, mechanical and thermal hyperalgesia of both hind feet were found in the PTX group, and the expression of NKCC1 and the number of activated microglia in dorsal horn tissue of spinal cord were increased. Compared with the PTX group, mechanical and thermal hyperalgesia were significantly improved in the PTX+EA group at day 14 and 21, and the expression levels of NKCC1 and Iba1 in dorsal horn tissue of spinal cord were decreased. There was no significant difference in KCC2 expression between the four groups. Conclusion Electroacupuncture can effectively relieve paclitaxol-induced neuropathic pain, which may be related to the inhibition of NKCC1 expression and microglia activation in spinal dorsal horn of rats.

Key words: electroacupuncture, paclitaxel, neuralgia, spinal cord dorsal horn, microglia, sodium-potassium-chloride symporters, neuropathic pain

中图分类号:

R245

图/表 8

图1 实验流程图

Fig. 1 Flow chart of the experimental outline

图2 大鼠各时间点体质量变化＊P<0.05，＊＊P<0.01；F组间=2.966，F时间=1 086.000＊＊，F交互=2.262＊，n=6。

Fig.2 Changes in body weight of rats at different time points

表1 各组大鼠不同时间点机械撤足阈值比较（n=6，g，$\bar{x} \pm s$）

Tab.1 Comparison of paw-withdrawal threshold at different time points between the four groups

组别	第-2天	第-1天	第1天	第3天	第5天	第7天	第14天	第21天
Vehicle组	23.72±4.05	24.11±3.09	24.85±3.90	22.85±3.90	22.46±4.57	22.85±3.90	24.11±3.09	24.11±3.09
PTX组	24.11±3.09	22.46±4.57	21.88±5.72	14.73±3.73^a	12.41±3.59^a	10.84±2.11^a	8.33±2.96^a	10.36±3.79^a
PTX+EA组	25.37±0.00	21.88±5.72	22.64±6.69	13.71±5.83^a	11.48±1.22^a	9.08±3.74^a	14.90±3.20^ab	20.91±6.91^b
PTX+Sham EA组	24.11±3.09	22.21±5.04	22.21±5.04	12.62±3.39^a	12.29±3.32^a	11.11±4.12^a	8.88±2.46^a	9.41±4.08^a

表2 各组大鼠不同时间点热缩足潜伏期比较（n=6，s，$\bar{x} \pm s$）

Tab.2 Comparison of paw-withdrawal latency at different time points of rats between the four groups

组别	第-2天	第-1天	第1天	第3天	第5天	第7天	第14天	第21天
Vehicle组	12.17±0.68	12.53±0.72	12.07±0.83	11.72±0.81	12.37±0.46	12.28±0.68	12.18±1.03	12.27±1.00
PTX组	12.78±0.29	12.59±1.07	12.25±0.82	8.98±0.24^a	7.03±0.77^a	5.57±0.37^a	5.21±1.19^a	5.65±0.36^a
PTX+EA组	12.60±0.60	12.55±0.83	11.14±1.26	9.16±0.38^a	6.70±0.50^a	6.00±0.77^a	10.31±1.15^b	11.47±0.58^b
PTX+Sham EA组	12.92±0.51	11.68±0.60	11.35±0.63	8.92±0.15^a	6.61±0.41^a	5.21±0.35^a	5.23±0.87^a	5.80±0.64^a

图3 Western blot检测各组大鼠脊髓背角中NKCC1、KCC2、Iba1蛋白表达 A：Vehicle组；B：PTX组；C：PTX+EA组；D：PTX+Sham EA组。

Fig.3 Western blot analysis of NKCC1, KCC2 and Iba1 protein expression levels in spinal dorsal horn of rats between the four groups

表3 各组大鼠脊髓背角组织中NKCC1、KCC2、Iba1相对蛋白表达量比较（n=3，$\bar{x} \pm s$）

Tab.3 Comparison of NKCC1, KCC2 and Iba1 relative protein expression levels in dorsal horn of spinal cord of rats between the four groups

组别	NKCC1	KCC2	Iba1
Vehicle组	1.00±0.07	1.00±0.06	1.00±0.23
PTX组	4.35±0.16^a	1.05±0.08	3.72±0.18^a
PTX+EA组	1.73±0.07^b	1.02±0.35	1.69±0.21^b
PTX+Sham EA组	3.76±0.63^a	0.99±0.16	3.70±0.32^a
F	70.440^＊＊	0.246	101.500^＊＊

表4 各组大鼠脊髓背角组织中NKCC1、KCC2、Iba1阳性染色面积百分比（n=3，%，$\bar{x} \pm s$）

Tab.4 Percentage of NKCC1, KCC2 and Iba1 positive staining area in spinal dorsal horn tissue of rats between the four groups

组别	NKCC1	KCC2	Iba1
Vehicle组	9.53±2.20	25.95±2.35	1.57±0.43
PTX组	21.23±1.45^a	25.48±1.43	8.22±1.07^a
PTX+EA组	10.04±1.01^b	25.61±1.94	2.03±0.61^b
PTX+Sham EA组	20.18±3.86^a	25.19±0.93	7.31±1.56^a
F	21.030^＊＊	0.099	34.940^＊＊

图4 大鼠脊髓组织NKCC1、KCC2和Iba1的表达变化（免疫荧光染色，×200）

Fig. 4 Expression changes of NKCC1, KCC2 and Iba1 in rat spinal cord（Immunofluorescence,×200）

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电针对紫杉醇诱导大鼠神经病理性疼痛的影响

The effect of electroacupuncture on paclitaxel-induced neuropathic pain in rats

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