miR-133b 修饰的间充质干细胞来源的外泌体在脑出血大鼠模型中的作用。
Role of Exosomes Derived from miR-133b Modified MSCs in an Experimental Rat Model of Intracerebral Hemorrhage.
机构信息
Department of Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, 188 Shizi Street, Suzhou, Jiangsu Province, 215006, China.
出版信息
J Mol Neurosci. 2018 Mar;64(3):421-430. doi: 10.1007/s12031-018-1041-2. Epub 2018 Feb 17.
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.
脑出血（ICH）死亡率和发病率高，预后较差，但迄今为止，有效的治疗方法仍然有限。微小 RNA（miRNA）是基因表达的重要调节剂，已被证明与各种中枢神经系统（CNS）疾病的发病机制有关。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 MSC 分泌的携带 miR-133b 的外泌体在 ICH 后大鼠脑组织中的作用。本研究采用成年雄性 Sprague-Dawley（SD）大鼠自体动脉血 ICH 模型。在 MSC 转染 miR-133b 模拟物 72 h 后，从 MSC 培养基中收集 miR-133b 修饰的 MSC 衍生外泌体，然后通过尾静脉注射给大鼠。通过实时 PCR 和 Western blot 分析分别检测各组大鼠分泌的外泌体和脑组织中 miR-133b 的水平，以及 RhoA、细胞外信号调节激酶（ERK1/2）磷酸化和 cAMP 反应元件结合蛋白（CREB）的水平。通过 TUNEL 和氟-金精 B 染色分别评估 miR-133b 对神经元凋亡和变性的影响。脑出血后 24 h 和 72 h，大鼠脑组织中 miR-133b 水平降低，72 h 时达到峰值。miR-133b 修饰的外泌体给药后 24 h，大鼠脑组织中 miR-133b 水平明显升高，脑出血后脑组织中凋亡和神经退行性神经元明显减少。Western blot 分析结果表明，miR-133b 修饰的外泌体处理显著抑制 RhoA 表达，激活脑出血后大鼠脑组织中的 ERK1/2/CREB。综上所述，我们的研究表明，MSC 衍生的 miR-133b 修饰的外泌体通过 miR-133b 介导的 RhoA 和 ERK1/2/CREB 发挥抗凋亡作用，对 ICH 后大鼠具有神经保护作用。
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