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背根神经节中γ-氨基丁酸能调制对颈部切口痛大鼠电针镇痛的作用

Contribution of GABAergic modulation in DRGs to electroacupuncture analgesia in incisional neck pain rats.

作者信息

Qiao Li Na, Yang Yong Sheng, Liu Jun Ling, Zhu Jiang, Tan Lian Hong, Shi Yi Nan, Zhu Bing, Rong Pei Jing

机构信息

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing, China.

Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences, Beijing, China,

出版信息

J Pain Res. 2019 Jan 17;12:405-416. doi: 10.2147/JPR.S180165. eCollection 2019.

Abstract

PURPOSE

Acupuncture therapy is effective for relieving postoperative pain. Our previous study showed that electroacupuncture (EA) at Futu (LI18) and Hegu (LI4)-Neiguan (PC6) could alleviate incisional neck pain, which was related with its effect in upregulating γ-aminobutyric acid (GABA) expression in cervical (C3-6) dorsal root ganglions (DRGs); but whether its receptor subsets GABAα2R and GABAR1 in C3-6 DRGs are involved in EA analgesia or not, it remains unknown.

MATERIALS AND METHODS

Seventy-five male Sprague Dawley rats were randomized to normal control, model, LI18, LI4-PC6, and Zusanli (ST36)-Yanglingquan (GB34) groups. The incisional neck pain model was established by making a longitudinal incision along the midline of the rats' neck, followed by repeated mechanical stimulation. EA was applied to bilateral LI18, LI4-PC6, or ST36-GB34 for 30 minutes at 4, 24, and 48 hours after operation. The thermal pain threshold of the neck was detected by a tail-flick unit, and the C3-6 DRGs were removed for assaying the immunoactivity of substance P (SP), GABAα2R, glial fibrillary acidic protein (GFAP; a marker of satellite glial cells [SGCs]), and GABAR1 and the expression of GABAα2R and GABAR1 mRNA and proteins using immunofluorescence, real-time PCR, and Western blotting, respectively.

RESULTS

The cervical thermal pain threshold was significantly lower in the model group than the normal group (<0.001), indicating hyperalgesia after neck incision, and was considerably increased in both EA-LI18 and LI4-PC6 groups (<0.001), but not in ST36-GB34 group compared with model group (>0.05). Immunofluorescence staining showed that GABAα2 R expressed on SP neurons, and GABAR1 on SGCs. EA of LI18 and LI4-PC6 markedly suppressed the modeling-induced upregulation of the immunoactivity of SP (<0.001 and <0.01, respectively) and GFAP (<0.01 and <0.001, respectively) and significantly reversed neck incision-induced downregulation of the expression of GABAα2R and GABAR1 mRNAs and proteins (<0.05).

CONCLUSION

EA of LI18 and LI4-PC6 has an analgesic effect in incisional neck pain rats, which is related to its effects in upregulating GABAergic inhibitory modulation on nociceptive peptidergic neurons and SGCs in cervical DRGs.

摘要

目的

针刺疗法对缓解术后疼痛有效。我们之前的研究表明,针刺扶突(LI18)和合谷(LI4)-内关(PC6)可减轻颈部切口疼痛,这与其上调颈段(C3-6)背根神经节(DRG)中γ-氨基丁酸(GABA)表达的作用有关;但其受体亚型GABAα2R和GABAR1在C3-6 DRG中是否参与电针镇痛尚不清楚。

材料与方法

75只雄性Sprague Dawley大鼠随机分为正常对照组、模型组、LI18组、LI4-PC6组和足三里(ST36)-阳陵泉(GB34)组。沿大鼠颈部中线做纵向切口建立颈部切口疼痛模型,随后进行重复机械刺激。术后4、24和48小时对双侧LI18、LI4-PC6或ST36-GB34进行电针刺激30分钟。用甩尾仪检测颈部热痛阈值,并取出C3-6 DRG,分别用免疫荧光、实时PCR和蛋白质印迹法检测P物质(SP)、GABAα2R、胶质纤维酸性蛋白(GFAP;卫星胶质细胞[SGC]的标志物)、GABAR1的免疫活性以及GABAα2R和GABAR1 mRNA及蛋白的表达。

结果

模型组颈部热痛阈值显著低于正常组(<0.001),表明颈部切口后出现痛觉过敏,与模型组相比,电针LI18组和LI4-PC6组热痛阈值均显著升高(<0.001),而ST36-GB34组无明显变化(>0.05)。免疫荧光染色显示,GABAα2R在SP神经元上表达,GABAR1在SGC上表达。电针LI18和LI4-PC6显著抑制模型诱导的SP免疫活性上调(分别为<0.001和<0.01)以及GFAP上调(分别为<0.01和<0.001),并显著逆转颈部切口诱导的GABAα2R和GABAR1 mRNA及蛋白表达下调(<0.05)。

结论

电针LI18和LI4-PC6对颈部切口疼痛大鼠具有镇痛作用,这与其上调颈段DRG中伤害性肽能神经元和SGC的GABA能抑制调节作用有关。

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