电针对炎性痛大鼠背根神经节卫星胶质细胞活化和 P2X7 受体表达的干预
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1.浙江中医药大学第三临床医学院针灸神经生物学实验室 浙江省针灸神经病学重点实验室, 杭州 310053; 2. 浙江中医药大学附属第三医院针灸科, 杭州 310053

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Effect of electroacupuncture on activation of satellite glial cells and P2X7 receptor in the DRG of rats with chronic inflammatory pain
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1.Department of Neurobiology and Acupuncture Research, the Third Clinical Medical College, Zhejiang Chinese Medical University, Key Laboratory of Acupuncture and Neurology of Zhejiang Province, Hangzhou 310053, China. 2. Department of Acupuncture and Moxibustion,the Third Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310005

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    摘要:

    目的 观察电针对慢性炎性痛大鼠患侧足跖机械痛阈(paw withdrawal thresholds,PWTs)以及背根神 经节(dorsal root ganglia, DRG)内卫星胶质细胞及其 P2X7 受体活化的影响,探讨电针抗大鼠炎性痛的外周机制。 方法 第一部分:将大鼠完全随机分为空白组(Con group,n = 12)、炎性痛组(CFA group,n = 12)、炎性痛+电针组 (CFA + EA group,n = 12)、炎性痛+假电针组(CFA + sEA group,n = 12)。 炎性痛大鼠于右后足掌侧正中皮下注射 完全弗氏佐剂(complete freud’s adjuvant,CFA)每只 0. 1 mL 构建模型;电针处理取穴“足三里”、“昆仑”,电针参数 为疏密波,频率 2 / 100 Hz,强度 0. 5-1-1. 5 mA(每个强度治疗 10 min),每日 1 次,连续 7 d。 检测造模前和造模后 1、 3、7、8、10、12、14 d 各组大鼠机械痛阈,采用免疫印迹法检测造模后 14 d 各组大鼠患侧 DRG 中神经胶质酸性蛋白 (glial fibrillary acidic protein,GFAP)及 P2X7 受体活化情况。 第二部分:将大鼠随机分为炎性痛+DMSO 组(CFA + DMSO group,n = 8)、炎性痛+P2X7R 抑制剂 A740003 组(CFA + A740003 group,n = 10)、炎性痛+电针+生理盐水组 (CFA + EA + NS group,n = 8)、炎性痛+电针+P2X7R 激动剂 BzATP 组(CFA + EA + BzATP group,n = 12),分别鞘内 注射相应药物。 分别于手术前、造模前、造模后和给药后各时点检测大鼠 PWTs 变化。 结果 (1)CFA 致炎性模型 大鼠痛阈显著下降(P < 0. 01),电针治疗后其机械痛阈显著升高(P < 0. 01)。 CFA 足底注射 14 d 后,大鼠患侧 L4 -6 DRG 中 GFAP、P2X7 蛋白表达显著增多(P < 0. 05);电针显著抑制大鼠患侧 L4-6 DRG 中 GFAP、P2X7 蛋白表 达(P <0. 05),而 CFA + sEA 组却无明显变化(P> 0. 05)。 ( 2) 与 CFA + DMSO 组比较,鞘内注射 P2X7 抑制剂 A740003 显著提高 CFA 大鼠机械痛阈(P < 0. 01);CFA + EA + BzATP 组大鼠 PWTs 明显低于 CFA + EA + NS 组 (P <0. 01)。 结论 抑制大鼠背根神经节卫星胶质细胞活化和 P2X7 受体表达参与电针抗慢性炎性痛,可能是电 针镇痛外周机制之一。

    Abstract:

    Objective To observe the effects of electroacupuncture (EA) on ipsilateral paw withdrawal thresholds (PWTs), activation of satellite glial cells, and the P2X7 receptor in the dorsal root ganglia (DRG) of rats with chronic inflammatory pain, and further explore the peripheral mechanism of EA against chronic inflammatory pain. Methods Part 1: Rats were randomly divided into the control group (Con group; n = 12), CFA-induced inflammatory pain group (CFA group; n = 12), CFA model plus EA treatment group (CFA + EA group; n = 12), and CFA model plus sham EA treatment group (CFA + sEA group; n = 12). The CFA inflammatory pain model was established by injecting 0. 1 mL CFA into the plantar of the rat right hind foot. EA at a 2 / 100 Hz alternative frequency and gradual intensity (0. 5, 1, and 1. 5 mA, 10 min each) was administered at acupoints “Zusanli” and “Kunlun” for 30 min per day for 7 days. PWTs were detected at 1, 3, 7, 8, 10, 12, and 14 days after CFA injection. Protein expression of glial fibrillary acidic protein (GFAP) and P2X7 receptor in DRG at day 14 post-CFA was measured by western blotting. Part 2: CFA rats were randomly divided into the CFA plus DMSO group (CFA + DMSO group; n = 8), CFA plus P2X7R agonist A740003 group (CFA + A740003 group; n = 10), EA plus normal saline group (CFA + EA + NS group; n = 8), and EA plus P2X7R agonist BzATP group (CFA + EA + BzATP group; n = 12), which were intrathecally administrated with DMSO, A740003, normal saline, and BzATP respectively. PWTs were measured at pre-surgery, pre-CFA, post-CFA, and post-intrathecal injection. Results Part 1: PWTs of CFA rats were decreased significantly (P < 0. 01) and EA significantly increased PWTs (P < 0. 01). CFA injection significantly increased the expression of GFAP and P2X7 receptor in the ipsilateral L4-6 DRG of rats (P< 0. 05). EA significantly reduced the expression of GFAP and P2X7 receptor in the L4-6 DRG of rats (P < 0. 05), but sham EA had no obvious effect (P > 0. 05). Part 2: Compared with the CFA + DMSO group, intrathecal injection of P2X7 receptor antagonist A740003 significantly increased PWTs of CFA rats (P < 0. 01). PWTs in the CFA + EA + BzATP group were significantly lower than those in the CFA + EA + NS group (P < 0. 01). Conclusions Electroacupuncture alleviates chronic inflammatory pain in rats, which might be related to the reduction of satellite glial cell activation and decrease of P2X7 receptor activation in the rat DRG. Inhibition of satellite glial cell activation and a decrease of P2X7 receptor expression contribute to EA treating chronic inflammatory pain, which may be one of the peripheral mechanisms of EA analgesia.

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周游,汪雯,蔡杨乾,杜俊英,邵晓梅,蒋永亮,刘伯一,高宏,梁宜,方剑乔.电针对炎性痛大鼠背根神经节卫星胶质细胞活化和 P2X7 受体表达的干预[J].中国实验动物学报,2020,28(4):494~502.

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  • 收稿日期:2020-04-27
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  • 在线发布日期: 2020-09-15
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