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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 investigate the effect of bone marrow mesenchymal stem cells (BMSCs) transplantation combined with electroacupuncture on the expression of aquaporin 4 (AQP-4) in the perihematomal zone of hematoma in rats with intracerebral hemorrhage. Methods Heparin and collagenase were injected into the caudate nucleus of adult SD rats to induce intracerebral hemorrhage. BMSCs were transplanted to hemorrhagic lesions on the 3rd postoperative day. After electroacupuncture stimulation (Ea-BMSCs), BMSCs transplantation group (BMSCs group), saline transplantation group (Control group) and electroacupuncture stimulation group (Ea group). The brain tissue samples were collected at 1, 3, 5, 7 and 14 days after treatment. The water content of brain tissue was detected by the wet and dry method. The expression of AQP-4 in peripheral blood of the mice was detected by immunohistochemistry and Western Blotting. The single factor variance LSD method for statistical analysis. Results AQP-4 was expressed in the pia mater, cerebral cortex, lateral ventricle choroid and peripheral brain region of hemorrhage, and co-expressed with GFAP on astrocytes. The water content of brain tissue in each experimental group began to rise on the third day, reaching the peak on the fifth day, and then gradually decreased. 3 AQP-4 protein in each group began to rise on the third day, reaching the peak on the fifth day, while there was no increase in the control group and the Ea group after reaching the peak value, but peak appeared on the 14th day in the BMSCs group and the Ea-BMSCs group . 4 On the 5th and 7th day, the expression of AQP-4 in Ea-BMSCs group and BMSCs group was lower than that in Control group (P <0.05), and there was no difference between Ea-BMSCs group and BMSCs group. On the 14th day, The expression of AQP-4 in Ea-BMSCs group was higher than that in Control group and BMSCs group (P <0.05). Conclusion Electroacupuncture combined with BMSCs transplantation can significantly reduce the water content of brain tissue, which may be related to the activation of AQP-4 activated by electroacupuncture stimulated by water-transport function.