骨髓间充质干细胞来源外泌体Zeb2在大鼠缺血性脑损伤后的表达及其意义

Expression and significance of Zeb2 in exosomes derived from bone marrow mesenchymal stem cells after ischemic brain injury in rats

目的探讨大鼠缺血性脑损伤后骨髓间充质干细胞及外泌体Zeb2基因的表达及意义。方法将18只SPF级SD雄性大鼠随机分为假手术组和模型组各9只,模型组采用右颈内动脉线栓法创建大鼠大脑中动脉缺血再灌注模型,假手术组仅结扎颈内动脉,不阻塞大脑中动脉,造成与模型组同样的手术损伤。根据Bederson方法进行神经行为学评分,并采用RT-PCR和Western blotting方法分别从RNA水平和蛋白水平检测骨髓间充质干细胞及其外泌体、血清外泌体Zeb2的表达。结果与假手术组相比,模型组即缺血再灌注处理后骨髓间充质干细胞、骨髓间充质干细胞来源的外泌体、血清外泌体中Zeb2 m RNA、Zeb2蛋白表达水平均呈明显上升趋势,两组差异具有统计学意义(P均<0.05)。结论大鼠缺血再灌注损伤后会上调骨髓间充质干细胞及外泌体Zeb2基因的表达,Zeb2具有重要的神经保护作用。

Objective To detect the expression of Zeb2 gene in bone marrow mesenchymal stem cells(BMSCs)and exosomes and to explore its significance after ischemic brain injury in rats.Methods Eighteen SPF grade and SD male rats were randomly divided into sham operation group and model group.The middle cerebral artery ischemia-reperfusion model was established by right internal carotid artery suture in the model group.In the sham operation group,the internal carotid artery was ligated without blocking the middle cerebral artery to result in the same surgical injury as that of the model group.According to Bederson method,the neurobehavioral score was performed,and we used the RT-PCR and Western blotting to test Zeb2 gene expression from the levels of RNA and protein in BMSCs,BMSCs exosomes and serum exosomes,respectively.Results Compared with sham operation group,the expression levels of Zeb2 mRNA and Zeb2 protein in BMSCs,BMSCs exosomes and serum exosomes of the model group after ischemia-reperfusion significantly increased,and the differences were statistically significant(all P<0.05).Conclusion The expression of Zeb2 gene in cells and exosomes is up-regulated after ischemia-reperfusion injury in rats,and Zeb2 has an important neuroprotective effect.