中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (6): 655-660.doi: 10.13418/j.issn.1001-165x.2016.06.012

• 实验研究 • 上一篇    下一篇

电针刺激联合BMSCs移植对脑出血大鼠出血灶周边区水通道蛋白4表达的影响

范光碧1, 先德海1, 邓莉2, 汤华军1, 张雁儒3, 杨朝鲜2     

  1. 1.西南医科大学人体解剖学教研室; 2.西南医科大学神经生物研究室,  四川   泸州   646000;
    3.郑州大学人体解剖学教研室,  郑州   450001
  • 收稿日期:2016-04-15 出版日期:2016-11-25 发布日期:2016-12-20
  • 通讯作者: 杨朝鲜,教授,E-mail: lyycx@foxmail.com
  • 作者简介:范光碧(1977-),女,四川泸州人,硕士,讲师,研究方向:脑损伤修复,E-mail: 792061264@qq.com
  • 基金资助:

    四川省应用基础研究计划项目(2013JY0075); 四川省教育厅重点项目(12ZA077)

Effects of electro-acupuncture combined with BMSCs transplantation on the expressions of AQP-4 in rats subjected to intracerebral hemorrhage

FAN Guang-Bi1, XIAN De-Hai1, DENG Li2,TANG Hua-Jun1,ZHANG Yan-ru3, YANG Chao-Xian2   

  1. 1.Department of Human Anatomy,SouthwestMedical University,Luzhou 646000,China; 2. Neurobiology Laboratory,SouthwestMedical University,Luzhou 646000,China; 3. Department of Human Anatomy,Zhengzhou University, Zhengzhou 450001, China
  • Received:2016-04-15 Online:2016-11-25 Published:2016-12-20
  • Contact: YANG Chao-xian, E-mail:lyycx@hotmail.com

摘要:

目的 探讨骨髓间充质干细胞(BMSCs)移植与电针刺激联合治疗对脑出血大鼠出血灶周边区水通道蛋白4(AQP-4)表达的影响。 方法 向成年SD大鼠右侧尾状核注射肝素和胶原酶诱导脑出血,并于术后第3天向出血灶移植BMSCs后施以电针刺激(Ea-BMSCs组),同时设置BMSCs移植组(BMSCs组)、生理盐水移植组(Control组)和电针刺激组(Ea组)。各组分别于治疗后1、3、5、7和14 d收集脑组织标本,采用干湿法检测脑组织水含量,免疫组化和Western Blotting检测出血灶周边区AQP-4的表达变化。采用单因素方差LSD法进行统计分析。   结果 ① AQP-4表达在软脑膜、大脑皮质、侧脑室脉络组织以及出血灶周边脑区,且与GFAP共表达于星形胶质细胞上。② 各实验组脑组织水含量在第3天开始升高,第5天达峰值,随后逐渐降低。③各组AQP-4蛋白在第3天开始升高,第5天达峰值,Control组和Ea组在达峰值后未见升高,而BMSCs组和Ea-BMSCs组在第14天再度出现峰值。 ④在第5天和第7天时,Ea-BMSCs组、BMSCs组AQP-4表达均低于Control组(P<0.05),Ea-BMSCs组与BMSCs组之间表达无差异;在第14天时,Ea-BMSCs组AQP-4表达高于Control组和BMSCs组(P<0.05)。   结论 电针刺激联合BMSCs移植可显著降低脑组织水含量,可能与电针刺激使移植后表达上调的AQP-4活化发挥水转运功能有关。

关键词: 脑出血, 电针刺激, 骨髓间充质干细胞, 水通道蛋白4

Abstract:

Objective To explore effects of electro-acupuncture stimulation combined with bone marrow mesenchymal stem cells( BMSCs) transplantation on the expression of Aquaporins-4(AQP-4)in rats after intracerebral hemorrhage. Methods ICH rats models were established by injecting the collagenase and heparin into the right caudate nucleus. After 3 days, the animals were treated with BMSCs transplantation and electro-acupuncture (Ea-BMSCs group), or dealt with BMSCs transplantation (BMSCs group), electro-acupuncture (Ea group) and physiological saline injection (Control group) respectively. The brain tissues of rats were collected at 1, 3, 5, 7 and 14 d after treatment. And then, immunohistochemical method and Western Blotting were used to detect expression of AQP-4, dry and wet ratio method was used to detect water content of brain. LSD method was performed for statistical analysis. Results ①AQP - 4 was found in the pia mater, cerebral cortex, choroid plexusand the area surrounding the hemorrhage, which had been expressed in astrocytes together with GFAP . ②Water content of each group begun to increase at 3d, quickly peaked at 5d, and then gradually decreased. ③AQP - 4 of the control group and Ea group begun to increase at 3d, peaked at 5d, and then gradually decreased similar to the water content; Nevertheless, it peaked again at 14d on the BMSCs group and Ea- BMSCs group. ④Expression of AQP-4 in the Ea-BMSCs group was lower than the control group at 5 d and 7 d(P<0.05); There was no difference when compared with the BMSCs group. At 14d, Expression of AQP-4 in the Ea-BMSCs group was higher than the control group and BMSCs group(P<0.05). Conclusion Electro-acupuncture combined with BMSCs transplantation can significantly reduce the brain water content,which is probably correlated with up-regulation AQP-4.

Key words: Intracerebral hemorrhage, Electro-acupuncture, Bone marrow mesenchymal stem cells, Aquaporins-4