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Effects of electroacupuncture at 2 and 100 Hz on rat type 2 diabetic neuropathic pain and hyperalgesia-related protein expression in the dorsal root ganglion

研究2 Hz 和100 Hz 电针对2 型糖尿病神经痛大鼠的镇痛作用以及对背根神经节致痛相关蛋白表达的影响

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Abstract

Objective

To investigate the analgesic effects of electroacupuncture (EA) at 2 and 100 Hz on type 2 diabetic neuropathic pain (DNP) and on the expressions of the P2X3 receptor and calcitonin gene-related peptide (CGRP) in the dorsal root ganglion (DRG).

Methods

Rat type 2 DNP was induced by a high calorie and high sugar diet fed for 7 weeks, plus a single intraperitoneal injection of streptozotocin (STZ) after 5 weeks. EA at 2 and 100 Hz was carried out once every day after 7 weeks for 7 consecutive days. Body weight, serum fasting insulin (FINS), fasting blood glucose (FBG), insulin sensitivity index (ISI), and paw withdrawal latency (PWL) were measured. The expressions of L4–L6 DRG P2X3 receptors and CGRP were assessed by immunofluorescence.

Results

Type 2 DNP was successfully induced as shown by the increased body weight, FINS, and FBG, as well as the reduced ISI and PWL. Expressions of P2X3 receptors and CGRP in L4–L6 DRGs increased. EA at both 2 and 100 Hz relieved type 2 DNP, but the analgesic effect of EA was stronger at 2 Hz. P2X3 receptor expression decreased in L4–L6 DRGs following EA at 2 Hz and in L5 and L6 DRGs following EA at 100 Hz. EA at both 2 and 100 Hz down-regulated CGRP overexpression in L4–L6 DRGs.

Conclusions

These findings indicate that EA at 2 Hz is a good option for the management of type 2 DNP. The EA effect may be related to its down-regulation of the overexpressions of the DRG P2X3 receptors and CGRP in this condition.

概要

目的

研究2 和100 Hz 电针对2 型糖尿病神经痛(DNP)大鼠的镇痛作用,以及对DNP 大鼠背根神经节(DRG)上P2X3 受体和降钙素相关基因肽(CGRP)表达的影响。

创新点

本研究首次选用2 型DNP 大鼠模型来研究电针治疗DNP 的优势频率。研究证明2 Hz 电针对2 型DNP 大鼠的镇痛作用以及对DRG 上高表达的P2X3 抑制作用优于100 Hz,为电针治疗DNP 及其频率选择提供科学依据和阐释。

方法

将雄性SD 大鼠分为对照组(腹腔注射柠檬酸钠缓冲液)和模型组(高脂高糖饲养联合腹腔注射35 mg/kg 链脲佐菌素(STZ))。模型组根据采用不同的电针频率分为以下三组:DNP 组、DNP+2 Hz 电针组、DNP+100 Hz 电针组。采用足底测试仪检测大鼠足跖热痛阈(PWL),用酶联免疫吸附测定(ELISA)试剂盒检测大鼠空腹胰岛素(FINS)含量,用免疫荧光法检测DRG 上的P2X3 受体和CGRP 的表达水平。

结论

2 和100 Hz 电针对2 型DNP 均有明显的镇痛作用,且2 Hz 电针对2 型DNP 的镇痛作用优于100 Hz 电针(图2)。免疫荧光实验结果显示,2和100 Hz 电针均能明显下调DRG 中P2X3 受体和CGRP 的高表达(图3 和4),且2 Hz 电针对DRG 中P2X3 受体的抑制作用优于100 Hz 电针(图3)。综上所述,2 Hz 电针是治疗2 型DNP较为理想的治疗手段。

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Acknowledgements

We would like to thank all the rats for giving their lives. We thank Qi-yang SHOU (Zhejiang Chinese Medical University, Hangzhou, China) for technical assistance.

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Authors and Affiliations

  1. Department of Neurobiology and Acupuncture Research, the Third Clinical Medical College, Zhejiang Chinese Medical University, Hangzhou, 310053, China

    Xiao-fen He, Jun-jun Wei, Sheng-yun Shou & Yong-liang Jiang

  2. Zhejiang Chinese Medical University, Hangzhou, 310053, China

    Jian-qiao Fang

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Correspondence to Jian-qiao Fang or Yong-liang Jiang.

Additional information

Project supported by the National Natural Science Foundation of China (No. 81303039), the Specialized Research Fund for the Doctoral Program of Higher Education (No. 20133322120001), the Zhejiang Postdoctoral Science Foundation (No. BSH1302083), the Zhejiang Province Top Key Discipline of Chinese Medicine-Acupuncture & Tuina (No. [2012]80), and the Key Science and Technology Innovation Team of Zhejiang Province (No. 2013TD15), China

ORCID: Yong-liang JIANG, http://orcid.org/0000-0002-9937-7398

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Xiao-fen HE, Jun-jun WEI, Sheng-yun SHOU, Jian-qiao FANG, and Yong-liang JIANG declare that they have no conflict of interest.

All institutional and national guidelines for the care and use of laboratory animals were followed.

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He, Xf., Wei, Jj., Shou, Sy. et al. Effects of electroacupuncture at 2 and 100 Hz on rat type 2 diabetic neuropathic pain and hyperalgesia-related protein expression in the dorsal root ganglion. J. Zhejiang Univ. Sci. B 18, 239–248 (2017). https://doi.org/10.1631/jzus.B1600247

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