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,先德海,邓莉,汤华军,张雁儒,杨朝鲜
中国临床解剖学杂志 ›› 2016, Vol. 34 ›› Issue (6) : 655-660.
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
Effects of electro-acupuncture combined with BMSCs transplantation on the expressions of AQP-4 in rats subjected to intracerebral hemorrhage
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。
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.
脑出血 / 电针刺激 / 骨髓间充质干细胞 / 水通道蛋白4
Intracerebral hemorrhage / Electro-acupuncture / Bone marrow mesenchymal stem cells / Aquaporins-4
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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）
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