Brain Endothelial Cell-Derived Exosomes Induce Neuroplasticity in Rats with Ischemia/Reperfusion Injury
Brain Endothelial Cell-Derived Exosomes Induce Neuroplasticity in Rats with Ischemia/Reperfusion Injury
复制标题
脑内皮细胞衍生的外泌体诱导缺血/再灌注损伤大鼠的神经可塑性
DOI：
10.1021/acschemneuro.0c00089
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发表时间：
2020-08-05
影响因子：
5
通讯作者：
Bai, Yulong
中科院分区：
文献类型：
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作者：
Gao, Beiyao;Zhou, Shaoting;Bai, Yulong
来源于脑内皮细胞的Exosomes在保护神经元免受缺氧损伤中发挥重要作用，但关于exosomes对脑可塑性的生物学作用和机制知之甚少。在这项研究中，通过超离心从啮齿动物脑内皮细胞（bEnd. 3细胞）中分离exosomes，分别在Exo组和对照组中的大脑中动脉闭塞/再灌注（MCAO/R）模型手术后2 h脑室内注射内皮细胞衍生的exosomes（EC-Exo）或PBS。Sham组大鼠接受相同的手术，但不接受缺血性手术。对MCAO/R后大鼠的运动功能进行评估，Exo组23天内的足错率显著低于对照组（p < 0.05）; Catwalk分析也显示两组之间的步态差异（p < 0.05）。MCAO/R后第28天，我们将大鼠安乐死，从大脑中取出运动皮层，然后使用GO和KEGG对基因进行测序，找到生物学术语和功能注释的转录组分析：通路富集显示，与对照组相比，Exo组的突触传递、突触可塑性调节和突触囊泡周期调节功能显著富集。与对照组相比，Exo组运动皮层突触蛋白I表达上调（p < 0.05），树突长度增加（p < 0.05）。我们测定了外泌体microRNA水平的含量，通过转录组分析，microRNA-126- 3 p的含量最高（TPM）。此外，microRNA-126- 3 p保护PC 12细胞免于凋亡并增加神经突生长，说明了外泌体如何在改变大脑可塑性中发挥作用的机制。这项研究表明，EC-Exo促进了MCAO/R模型中的功能运动恢复，外泌体对于缺血性脑损伤中突触功能的重建至关重要，并且来自EC-Exo的microRNA-126- 3 p可以作为神经损伤的治疗。
Exosomes derived from the cerebral endothelial cells play essential roles in protecting neurons from hypoxia injury, but little is known regarding the biological effects and mechanisms of exosomes on brain plasticity. In this study, exosomes were isolated from rodent cerebral endothelial cells (bEnd.3 cells) by ultracentrifugation, either endothelial cell-derived exosomes (EC-Exo) or PBS was injected intraventricularly 2 h after the middle cerebral artery occlusion/reperfusion (MCAO/R) model surgery in the Exo group and control group, respectively. Sham group rats received the same surgical but not ischemic procedure. We evaluated the motor function of rats after MCAO/R, and the foot-fault rate of the Exo group was significantly lower than that of the control group within 23 days (p < 0.05); the Catwalk analysis also showed gait difference between two groups (p < 0.05). On day 28 after MCAO/R, we euthanized the rats, removed the motor cortex from the brain, and then sequenced the genes by using GO and KEGG to find transcriptome analysis of biological terms and functional annotations: The pathway enrichment revealed that the function of synaptic transmission, regulation of synaptic plasticity, and regulation of synaptic vesicle cycle was significantly enriched with the Exo group than control group. Furthermore, the upregulation of synapsin-I expression in the motor cortex (p < 0.05) as well as the increase of the length of the dendrites were found in the Exo group (p < 0.05) than the control group. We determined the content of exosome microRNA levels, and microRNA-126-3p was the highest (TPM) by transcriptome analysis. Moreover, the microRNA-126-3p protected PC12 cells from apoptosis and increased neurite outgrowth, illustrating the mechanism of how exosomes play a role in altering brain plasticity. This study demonstrated that EC-Exo promoted functional motor recovery in the MCAO/R model, exosomes were critical for the reconstruction of synaptic function in ischemic brain injury, and microRNA-126-3p from EC-Exo could serve as a treatment for nerve damage.