电针百会、四关穴对大鼠局灶性脑缺血再灌注大鼠大脑皮质中Tax1结合蛋白1表达的影响

doi:10.3969/j.issn.1006-9771.2017.12.002

《中国康复理论与实践》 ›› 2017, Vol. 23 ›› Issue (12): 1372-1379.doi: 10.3969/j.issn.1006-9771.2017.12.002

电针百会、四关穴对大鼠局灶性脑缺血再灌注大鼠大脑皮质中Tax1结合蛋白1表达的影响

马宏梅², 蒋锦², 詹剑², 李琼莉², 张莹², 秦文熠, 罗勇²

1.重庆医科大学附属第一医院,a.神经内科,b.中西医结合科,重庆市 400016;
2.重庆市神经病学重点实验室,重庆市 400016。

收稿日期:2017-07-18 修回日期:2017-11-08 出版日期:2017-12-25 发布日期:2017-12-28
通讯作者: 罗勇(1965-),男,汉族,重庆市人,博士,博士生导师,研究员,主要研究方向：脑血管病的损伤及保护机制。E-mail: luoyong1998@163.com。
作者简介:马宏梅(1990-),女,布依族,贵州龙里县人,硕士研究生,主要研究方向：针刺治疗脑血管病的保护机制。
基金资助:
1.国家自然科学基金项目(No. 30470606; No. 81403243); 2.重庆市渝中区科技计划项目(No. 20160102); 3.重庆市卫生局中医药科技项目(No. 2012-2-128)

Effect of Electroacupuncture at Baihui and Siguan on Expression of Tax1-binding Protein 1 in Cerebral Cortex in Focal Cerebral Ischemia-Reperfusion Rats

MA Hong-mei², JIANG Jin², ZHAN Jian², LI Qiong-li², ZHANG Ying², QIN Wen-yi, LUO Yong²

1.a. Department of Neurology, b. Department of Integrated Chinese and Western Medicine, the First Affiliated Hospital of Chonqing Medical University, Chongqing 400016, China;
2.Chongqing Key laboratory of Neurology, Chongqing 400016, China

Received:2017-07-18 Revised:2017-11-08 Published:2017-12-25 Online:2017-12-28
Contact: LUO Yong. E-mail: luoyong1998@163.com

摘要/Abstract

摘要： 目的观察电针百会、四关穴对大鼠局灶性脑缺血再灌注大鼠大脑皮质中Tax1结合蛋白1(TAX1BP1)表达的影响,探讨电针抑制核因子-κB (NF-κB)信号通路的激活,发挥脑保护作用的可能机制。方法 105只健康雄性Sprague-Dawley大鼠随机分为假手术组、模型组和电针组,每组再分为缺血2 h后再灌注6 h、12 h、24 h、48 h、72 h五个亚组。改良Longa线栓法制备右侧大脑中动脉梗死再灌注模型。电针组给予电针百会穴和病侧四关(合谷/太冲)穴。检测各组神经功能,缺血区大脑皮质中TAX1BP1蛋白的表达情况、TAX1BP1阳性细胞数、锌指蛋白A20和胞核NF-κB p65蛋白的表达情况。结果假手术组无神经功能缺损,与模型组相比,电针组在局灶性脑缺血2 h再灌注48 h、72 h时神经功能评分降低(P<0.05)。与假手术组相比,模型组在再灌注12 h、24 h、48 h时TAX1BP1蛋白表达增加(P<0.05);与模型组相比,电针组在再灌注12 h、24 h、48 h、72 h时TAX1BP1蛋白表达进一步增加(P<0.05),再灌注24 h为表达高峰。在再灌注24 h后,与假手术组相比,模型组A20表达、TAX1BP1阳性细胞数、胞核NF-κB p65表达增加(P<0.05);与模型组相比,电针组的A20表达、TAX1BP1阳性细胞数进一步增加(P<0.05),胞核NF-κB p65蛋白表达减少(P<0.05)。免疫荧光结果显示,TAX1BP1蛋白主要表达在胞浆,电针组TAX1BP1与A20共表达在胞浆。免疫组化结果显示,模型组NF-κB p65主要表达在胞核,电针组主要表达在胞浆。结论电针抑制大鼠局灶性脑缺血再灌注后NF-κB信号通路的激活,促进大鼠神经功能恢复,其机制可能是通过上调TAX1BP1蛋白的表达,从而发挥脑保护作用。

关键词: 脑缺血再灌注, 电针, Tax1结合蛋白1, 锌指蛋白A20, 核因子-κB p65, 大鼠

Abstract: Objective To observe the effect of electroacupuncture (EA) at Baihui (GV20) and Siguan (Hegu/LI4 and Taichong/LR3) of affected side on expression of Tax1-binding protein 1 (TAX1BP1) in cerebral cortex in focal cerebral ischemia-reperfusion rats, so as to investigate its protective mechanism in inhibiting nuclear factor kappa B (NF-κB) signaling pathway and promoting neurobehavioral recovery. Methods A total of 105 Sprague-Dawley rats were randomly assigned into sham group, model group and EA group. Each group was randomly assigned into reperfusion six hours, twelve hours, 24 hours, 48 hours, 72 hours groups after two hours of ischemia. The model was established by right middle cerebral artery occlusion and reperfusion. EA group received electroacupuncture at Baihui and left Siguan (Hegu and Taichong) acupoints. Neurobehavioral evaluation, TAX1BP1 protein expression, TAX1BP1 positive cell count, zink finger protein A20 expression, and nuclear NF-κB p65 protein expression were tested in each group. Results There was no neurological deficit in the sham group. Compared with the model group, the neurological scores at 48 hours, 72 hours after reperfusion decreased in EA group (P<0.05). Compared with the control group, the TAX1BP1 expression at twelve hours, 24 hours and 48 hours after reperfusion increased in the model group (P<0.05), and further increased at twelve hours, 24 hours, 48 hours and 72 hours after reperfusion in EA group (P<0.05), and peaked at 24 hours after reperfusion. Compared with the sham group, the expression of A20 and NF-κB p65, and the number of TAX1BP1 positive cells increased in the model group (P<0.05), and the expression of A20 and the number of TAX1BP1 positive cells further increased, (P<0.05) and the expression of NF-κB p65 decreased in EA group (P<0.05) at 24 hours after reperfusion. Immunofluorescence labeling indicated that TAX1BP1 protein primarily expressed in the cytoplasm, TAX1BP1 protein and A20 protein co-expressed in the cytoplasm. Immunohistochemistry showed indicated that NF-κB p65 mainly expressed in the nucleus in the model groupr, and mainly expressed in the cytoplasm in the EA group. Conclusion Electroacupuncture could significantly inhibit neuronal NF-κB signaling pathway and promote neurobehavioral recovery in focal cerebral ischemia-reperfusion, which may related with up-regulating TAX1BP1 protein expression.

Key words: cerebral ischemia-reperfusion, electroacupuncture, Tax1-binding protein 1, zink finger protein A20, nuclear factor kappa B p65, rats

中图分类号:

R743.3

马宏梅, 蒋锦, 詹剑, 李琼莉, 张莹, 秦文熠, 罗勇. 电针百会、四关穴对大鼠局灶性脑缺血再灌注大鼠大脑皮质中Tax1结合蛋白1表达的影响[J]. 《中国康复理论与实践》, 2017, 23(12): 1372-1379.

MA Hong-mei, JIANG Jin, ZHAN Jian, LI Qiong-li, ZHANG Ying, QIN Wen-yi, LUO Yong. Effect of Electroacupuncture at Baihui and Siguan on Expression of Tax1-binding Protein 1 in Cerebral Cortex in Focal Cerebral Ischemia-Reperfusion Rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2017, 23(12): 1372-1379.

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电针百会、四关穴对大鼠局灶性脑缺血再灌注大鼠大脑皮质中Tax1结合蛋白1表达的影响

Effect of Electroacupuncture at Baihui and Siguan on Expression of Tax1-binding Protein 1 in Cerebral Cortex in Focal Cerebral Ischemia-Reperfusion Rats

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