Role of Exosomes Derived from miR-133b Modified MSCs in an Experimental Rat Model of Intracerebral Hemorrhage
Role of Exosomes Derived from miR-133b Modified MSCs in an Experimental Rat Model of Intracerebral Hemorrhage
复制标题
miR-133b 修饰的 MSC 衍生的外泌体在实验性大鼠脑出血模型中的作用
DOI：
10.1007/s12031-018-1041-2
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发表时间：
2018-03-01
影响因子：
3.1
通讯作者：
Chen, Gang
中科院分区：
文献类型：
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作者：
Shen, Haitao;Yao, Xiyang;Chen, Gang
脑出血（Intracerebral hemorrhage，ICH）因其高病死率和高致残率而导致预后不良，但目前有效的治疗方法仍有限。microRNAs（miRNAs）是基因表达的重要调节因子，与多种中枢神经系统疾病的发病机制密切相关。外泌体被认为是细胞间通讯的载体，主要由间充质基质细胞（MSC）分泌。本研究探讨了miR-133 b在大鼠ICH后由MSC分泌的外泌体递送到脑组织中的作用。本研究使用成年雄性Sprague-Dawley（SD）大鼠自体动脉血ICH模型。在MSC中转染miR-133 b模拟物后72 h，从MSC的培养基中收集miR-133 b修饰的MSC衍生的外泌体，然后通过尾静脉注射至大鼠。分别采用实时定量PCR和Western blot检测各组大鼠分泌的exosomes和脑组织中miR-133 b的水平，以及RhoA、细胞外信号调节激酶（ERK 1/2）磷酸化和cAMP反应元件结合蛋白（CREB）的水平。TUNEL法检测miR-133 B对神经元凋亡的影响，fluoro-jade B染色检测miR-133 b对神经元变性的影响。大鼠脑出血后24 h脑组织中miR-133 b水平降低，72 h达到高峰。注射miR-133 b修饰的exosome后24 h，脑出血后脑组织中miR-133 b水平显著升高，而凋亡和神经变性神经元明显减少。Western blot分析结果显示，miR-133 b修饰的exosomes处理可显著抑制脑出血后脑组织中RhoA的表达，并激活ERK 1/2/CREB。总之，我们的研究表明，来源于miR-133 b修饰的MSCs的exosomes在ICH后大鼠中表现出miR-133 b介导的RhoA和ERK 1/2/CREB的抗凋亡作用的神经保护作用。
Intracerebral hemorrhage (ICH) has poor outcomes due to high mortality and morbidity, but until now, the effective treatments remain limited. MicroRNAs (miRNAs) are vital regulators of gene expression and demonstrated to be linked to the pathogenesis of various central nervous system (CNS) diseases. Exosomes are considered as cell-to-cell communication vectors and secreted largely by mesenchymal stromal cells (MSCs). The present study investigated the role of miR-133b delivered by exosomes secreted from MSCs to brain tissues in rats after ICH. An autologous arterial blood ICH model in adult male Sprague–Dawley (SD) rats was used in this study. At 72 h after transfection with miR-133b mimics in MSCs, miR-133b-modified MSC-derived exosomes were collected from medium of MSCs and then injected to rats via tail vein. The levels of miR-133b in secreted exosomes and brain tissues of rats in various groups and the levels of RhoA, phosphorylations of extracellular signal regulating kinase (ERK1/2), and cAMP response element-binding protein (CREB) were detected by real-time PCR and western blot analysis, respectively. The effects of miR-133b on neuronal apoptosis and degeneration were respectively evaluated by TUNEL and fluoro-jade B staining. The miR-133b levels were reduced in brain tissues of rats at 24 h and peaked at 72 h after ICH. At 24 h after miR-133b-modified exosome administration, the level of miR-133b was significantly increased, while the apoptotic and neurodegenerative neurons were obviously reduced in brain tissues after ICH. The results of western blot analysis showed that miR-133b modified exosomes treatment remarkably suppressed RhoA expression and activated ERK1/2/CREB in brain tissues after ICH. Collectively, our investigation suggested that exosomes derived from miR-133b modified MSCs exhibited neuroprotective role for anti-apoptotic effect of miR-133b mediating RhoA and ERK1/2/CREB in rats after ICH.