不同时间电针干预痛厌恶大鼠焦虑行为的前扣带皮层机制研究
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浙江中医药大学第三临床医学院康复医学院 浙江省针灸神经病学研究重点实验室,杭州 310053


Effect of electroacupuncture intervention on anxiety behaviors of pain-aversion rats and the mechanism of the anterior cingulate cortex
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Third Clinical Medical College and Rehabilitation Medical College of Zhejiang Chinese Medical University, Key Laboratory of Acupuncture and Neurology of Zhejiang Province, Hangzhou 310053, China

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    摘要:

    目的 探究电针干预对痛厌恶大鼠焦虑行为的缓解效应和其对前扣带皮层( anterior cingulate cortex,ACC)N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)和 γ-氨基丁酸(γ-aminobutyric acid,GABA)受体的影响。 方法 所有健康大鼠完全随机分为空白(NS)组、模型(C-CPA)组、常规电针(EA1)组、预电针(EA2)组,建立完全弗氏佐剂诱导的条件位置回避(complete Freund’ s adjuvant induced conditioned place avoidance,C-CPA)痛厌恶大鼠模型。 EA1 组于模后介入电针,EA2 组于造模前 3 d 介入电针,观察大鼠的机械缩足阈( paw withdrawal thresholds,PWTs)和焦虑行为;用免疫印迹法检测 ACC 内 GABA 的受体 GABAAR、GABABR,NMDA 的受体 NR2A、 NR2B 蛋白含量。 结果 与 NS 组相比,C-CPA 组、EA1 组和 EA2 组的逃避分数(CPA scores)明显减少(P< 0. 01), 且各组间无显著差异,说明痛厌恶大鼠模型造模成功。 造模前各组大鼠 PWTs 无显著变化;造模后,C-CPA 组大鼠出现痛觉过敏(P< 0. 01),焦虑情绪行为(P< 0. 01);EA1 组和 EA2 组痛觉过敏(P< 0. 01)和焦虑情绪行为均缓解 (P< 0. 05),且两组间无显著差异(P> 0. 05)。与 NS 组比较,C-CPA 组、EA1 组和 EA2 组患侧 ACC 内 GABAAR 蛋白表达显著增多(P< 0. 05);与 C-CPA 组比较,EA1 组患侧 ACC 内 GABAAR 蛋白表达无显著差异(P> 0. 05),EA2 组明显减少(P< 0. 05),但 EA1 组与 EA2 组无显著差异(P> 0. 05)。 患侧 ACC 中 GABABR、NR2A 和 NR2B 蛋白表达各组间比较均无差异(P> 0. 05)。 结论 不同时间电针干预均可减缓 C-CPA 痛厌恶模型大鼠疼痛和焦虑行为, 其中预电针的疗效可能与抑制患侧 ACC 内 GABAAR 的蛋白表达相关。

    Abstract:

    Objective To explore the relieving effect of electroacupuncture on the anxiety behaviors of pain- aversion rats and its effects on N-methyl-D-aspartate (NMDA) and γ-aminobutyric acid (GABA) receptors in the anterior cingulate cortex (ACC). Methods Healthy rats were randomly divided into four groups: control group ( NS group), model group (C-CPA group), normal electroacupuncture group (EA1 group), and pre-electroacupuncture group (EA2 group). The rats of the C-CPA, EA1, and EA2 groups were established pain-aversion models that were injected subcutaneously in the hind paw with complete Freund’ s adjuvant (CFA) and were acclimatized to one side of the cage apparatus to induce conditioned place avoidance (CPA) responses. The rats in the EA1 group underwent intervention with electroacupuncture after modeling, while those in the EA2 group underwent intervention 3 days before modeling. The paw withdrawal thresholds ( PWTs) and anxiety behaviors of rats were observed. Western blotting was used to detect the expression of GABA receptor ( GABAAR, GABABR) and NMDA receptor ( NR2A, NR2B) in the ACC. Results Compared with the NS group, the CPA scores of the C-CPA, EA1, and EA2 groups were decreased (P< 0. 01), and there was no significant difference among the groups, which indicated that the establishment of the pain aversion rat model was successful. Moreover, there was no significant changes in PWTs before modeling. After modeling, rats in the C-CPA group showed hyperalgesia (P< 0. 01) and pain-related anxiety behaviors (P< 0. 01). PWTs and pain-related anxiety behaviors of the rats in the EA1 and EA2 groups were significantly relieved (P< 0. 01 and P< 0. 05, respectively), and there were no significant differences between the two groups (P> 0. 05). Compared with the NS group, GABAAR protein expression in the ipsilateral ACC of the C-CPA, EA1, and EA2 groups was significantly increased (P< 0. 05). There was no significant difference in GABAAR expression in the ipsilateral ACC between the C-CPA and EA1 groups (P> 0. 05), but GABAAR expression in the ipsilateral ACC of the EA2 group was decreased compared with that of the C-CPA group (P< 0. 05). There was no significant difference between the EA1 and EA2 groups (P> 0. 05). GABABR, NR2A, and NR2B expression in the ipsilateral ACC exhibited no differences among the NS, C-CPA, EA1, and EA2 groups (P> 0. 05). Conclusions Both normal- and pre-electroacupuncture intervention could relieve the pain and pain-related anxiety behaviors of C-CPA pain aversion models. The efficacy of pre-electroacupuncture intervention may be related to inhibition of GABAAR expression in the ipsilateral ACC.

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沈科展,吴学文,何淑峥,孙鑫,周之东,朱雨鑫,陈扬,方剑乔,房军帆,杜俊英.不同时间电针干预痛厌恶大鼠焦虑行为的前扣带皮层机制研究[J].中国实验动物学报,2021,29(3):332~342.

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  • 收稿日期:2020-12-07
  • 在线发布日期: 2021-08-13
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