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 ofAquaporins-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.