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Effect of electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) on the expression of EphB2 protein in cortex around cerebral infracted area of rat

电针肝俞、肾俞对大鼠脑梗死灶周围皮层EphB2 蛋白表达的影响

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Abstract

Objective

To observe the effect of electroacupuncture (EA) on the expression of erythropoie-tin-producing hepatocyte receptor B2 (EphB2) in the cortex around the infracted area of middle cerebral artery occlusion (MCAO) rats at different timing, and to reveal the possible mechanism of acupuncture in the treatment of cerebral ischemia.

Methods

A total of 180 male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, with 45 rats in each group. Rats in each group were further divided into three subgroups: postoperative 3 d, postoperative 14 d and postoperative 21 d groups, with 15 rats in each subgroup. The MCAO model was made by the modified occlusion method. The neurological function score, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, immunohistochemistry assay, immunofluorescence double labeling method and Western blot were used to detect the corresponding indicators.

Results

The neurological impairment of rats was most obvious at postoperative 3 d, and then gradually improved with time, which was more significant in the acupoint group (P<0.05). The change of infarcted volume was consistent with the neurological function impairment. The number of EphB2 positive cells (EphB2+) around the infarcted area was decreased significantly at postoperative 3 d, and then gradually improved with time, which returned to the same level as that in the sham operation group at postoperative 21 d. The increase was most significant in the acupoint group (P<0.05), and the positive cell number was higher than that in the sham operation group (P<0.01). Western blot and immunohistochemistry results were basically consistent. Immunofluorescence displayed that EphB2+ and postsynaptic density-95 positive (PSD-95+) were co-expressed, after the MCAO operation, in the cortical neuron around the infracted area, and the number of co-expressing cells was increased gradually with time, which was most significant in the acupoint group (P<0.05).

Conclusion

Electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function and cerebral infarcted volume ratio of MCAO rats, which may be related to the activation of EphB2 expression in cortex around the infracted area and the promotion of synaptic remodeling.

摘要

目的

观察电针对大脑中动脉梗塞(MCAO) 模型大鼠不同时间点梗死灶周围皮层促红细胞生成素产生肝 细胞受体B2 (EphB2)表达的影响, 以期揭示针刺治疗脑缺血病的可能作用机制。

方法

将180 只雄性Sprague-Dawley (SD)大鼠随机分为假手术组、模型组、穴位组和非穴位组, 每组45 只; 各组又分为术后3 d, 14 d 及21 d 三个亚组, 每组15 只。采用改良线栓法复制MCAO 大鼠模型, 采用神经功能评分、2,3,5-氯化三苯基四氮唑(TTC) 染色、免疫组化法、免疫荧光双标法及Western blot 法进行相应指标检测。

结果

大鼠术后神经功能缺损在3 d 时最明显, 之后随着时间延长逐渐改善, 其中以穴位组更显著(P<0.05); 且梗死灶体积变化与神经功能缺损基本 一致。术后3 d 梗死灶周围EphB2 阳性(EphB2+)细胞数明显减少, 随着时间推移表达逐渐增加,至21 d 恢复至假手 术组水平, 其中以穴位组增加最明显(P<0.05), 且阳性细胞数高于假手术组(P<0.01)。Western blot 与免疫组化 检测结果基本一致。免疫荧光显示大鼠术后EphB2+与突触后致密物-95 阳性(PSD-95+)在梗死灶周围皮层神经细胞 上共表达, 且随时间延长共表达细胞数逐渐增加, 以穴位组增加最明显(P<0.05)。

结论

电针肝俞、肾俞能明显 改善MCAO 大鼠神经功能及脑梗死体积比, 可能与激活梗死灶周围皮层EphB2 表达, 促进突触重塑有关。

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Acknowledgments

This work was supported by the Open Fund for Colleges and Universities Innovation Platform of Hunan Province (湖南省高校创新平台开放基金, No. 14K070); Key Project of Hunan Province Administration of Traditional Chinese Medicine (湖南省中医药管理局重点项目, No. 201310); Innovation Fund Project for Graduate Student of Hunan Province ( 湖南省研究生创新基金, No. CX2014B360).

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Authors and Affiliations

  1. Department of Acupuncture and Moxibustion, the First Hospital of Hunan University of Chinese Medicine, Changsha, 410007, China

    Hong-liang Li  (李洪亮)

  2. School of Acupuncture, Moxibustion & Tuina, Hunan University of Chinese Medicine, Changsha, 410007, China

    Juan Xiang  (向娟), Li-zhi Ouyang  (欧阳里知), Guo Chen  (陈果), Zhi-hui Xue  (薛智慧), Kang-sheng Long  (龙抗胜) & Tie-lang Li  (李铁浪)

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  1. Hong-liang Li  (李洪亮)
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Correspondence to Tie-lang Li  (李铁浪).

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Li, Hl., Xiang, J., Ouyang, Lz. et al. Effect of electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) on the expression of EphB2 protein in cortex around cerebral infracted area of rat. J. Acupunct. Tuina. Sci. 15, 14–21 (2017). https://doi.org/10.1007/s11726-017-0968-0

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  • DOI: https://doi.org/10.1007/s11726-017-0968-0

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