回药扎里奴思方联合骨髓间充质干细胞移植对脑缺血再灌注大鼠脑水肿及水通道蛋白4表达的影响

Effects of Hui medicine Zhali Nusi Recipe(ZLNS) combined with bone marrow mesenchymal stem cells transplantation on brain edema and the expression of aquaporin 4 in rats with cerebral ischemia reperfusion injury

目的:观察扎里奴思方联合BMSCs移植对MCAO模型大鼠脑水肿及AQP4表达的影响。方法:SD大鼠随机分为假手术组、模型组、扎方组、移植组和联合组;线栓法制备大鼠MCAO模型,体外全骨髓贴壁筛选法培养及扩增BMSCs;大鼠灌胃给药,BMSCs悬浮液经颈内动脉移植入脑;移植后1、3、7、14d取材,评价神经功能积分,干湿重法测脑含水量、免疫组化法测AQP4表达变化。结果:模型大鼠神经功能评分较假手术组显著降低(P<0.01),脑含水量、AQP4表达显著增高(P<0.01);与模型组比较,扎方组1、3、14d,移植组3、7、14d及联合各组神经功能评分增高(P<0.01,P<0.05),扎方、移植及联合组各3、7、14d脑含水量及AQP4表达降低(P<0.01);与移植组比较,扎方组7d及联合组1、3、14d神经功能评分增高(P<0.01,P<0.05),扎方组1d、联合组1、14d脑含水量降低(P<0.05),联合各组AQP4表达降低(P<0.01);扎方组与联合组比较,联合组上述指标均明显改善,以7、14d变化显著(P<0.01);同组间比较,均以7d组变化显著,14d有显著改善(P<0.01)。结论:脑缺血后神经功能损伤,脑水肿和AQP4表达均呈先增后减变化趋势;扎里奴思方和BMSCs移植均可不同程度改善脑缺血后神经功能损伤和脑水肿程度,以二者联合应用作用显著,其机制可能与干预AQP4动态表达有关。

Objective： To observe the effect of Hui medicine Zhali Nusi Recipe（ZLNS） combined with bone marrow mesenchymal stem cells（BMSCs） transplantation on brain edema and aquaporin 4（AQP4） expression in rats with cerebral ischemia reperfusion injury（CIRI）. Methods： SD rats were randomly divided into the Sham-operated group, model group, ZLNS group, BMSCs transplantation group and ZLNS combined with BMSCs group. Middle cerebral artery occlusion（MCAO） model was established with nylon thread, and BMSCs were cultured and amplified by the whole bone marrow adherence method; The drug administration was performed by gavage, and BMSCs suspension solution were transplanted into brain through carotid artery. The rats were sacrificed to take out the brain tissue at the 1st, 3rd, 7th, 14 th day after transplantation, respectively. The neurological score（NS） was used to evaluate the neurological function. And, the brain water content（BWC） was detected by wet and dry weight, and the expression of AQP4 were detected by immunohistochemical method. Results： The NS in model groups were significantly lower than that in the sham-operated group（P〈0.01）, while the expressions of BWC and AQP4 in the model groups significantly higher than those in the sham-operated group（P〈0.01）. Compared with the model groups, NS at the 1st, 3rd, 14 th day in ZLNS group, NS at the 3rd, 7th, 14 th day in transplantation groups and in all combination groups were increased（P〈0.01, P〈0.05）, and BWC and AQP4 at the 3rd, 7th, 14 th day in ZLNS, transplantation and combination groups were significantly decreased（P〈0.01）. Compared with the transplantation group, NS at the 7th day in ZLNS groups and NS at the 3rd, 14 th day in combination groups were increased（P〈0.01, P〈0.05）, the BWC at the 1st day in ZLNS group and BWC at the 1st and 14 th day in combination groups were decreased（P〈0.05）, the expression of AQP4 in each combination groups were obviously decreased（P〈0.01）. Compared with the ZLNS groups, all the indicators in each combination groups were improved, especially at the 7th, 14 th day（P〈0.01）. The comparison among the ZLNS groups showed that the changes were significantly observed at the 7th day, and the improvement was at the 14 th day（P〈0.01）. Conclusion： After cerebral ischemia, the tendency of NS, BCW and the expression of AQP4 shows an increase at first and then decrease. Both ZLNS and BMSCs transplantation could improve the nerve function deficit and brain edema in different extent, and their combination could show superior effect, which may be related to the regulation of dynamic expression of AQP4.