Key Laboratory of Modern Toxicology (NJMU), Ministry of Education, Department of Toxicology, School of Public Health, Nanjing Medical University, 818 Tianyuan East Road, Nanjing, Jiangsu, 211166, China; Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, 210029, China.
Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu, 210029, China.
Exp Cell Res. 2021 Aug 1;405(1):112635. doi: 10.1016/j.yexcr.2021.112635. Epub 2021 May 26.
Exosomes have been shown to have therapeutic potential for cerebral ischemic diseases. In this study, we investigated the neuroprotective effects of normoxic and hypoxic bone marrow mesenchymal stromal cells-derived exosomes (N-BM-MSCs-Exo and H-BM-MSCs-Exo, respectively) on oxygen-glucose deprivation (OGD) injury in mouse neuroblastoma N2a cells and rat primary cortical neurons. The proportions of dead cells in N2a and primary cortical neurons after OGD injury were significantly increased, and N-BM-MSCs-Exo (40 μg/ml) could reduce the ratios, noteworthily, the protective effects of H-BM-MSCs-Exo (40 μg/ml) were more potent. Western blotting analysis indicated that N-BM-MSCs-Exo decreased the expression of NLRP3, ASC, Caspase-1, GSDMD-N, cleaved IL-1β and IL-18 in N2a cells. However, H-BM-MSCs-Exo (40 μg/ml) was more powerful in inhibiting the expression of these proteins in comparison with N-BM-MSCs-Exo. Similar results were obtained in primary cortical neurons. Immunofluorescence assays showed that after N-BM-MSCs-Exo and H-BM-MSCs-Exo treatment, the co-localization of NLRP3, ASC, Caspase-1 and the GSDMD translocation from the nucleus to the cytoplasm and membrane after OGD injury were reduced in N2a cells and primary cortical neurons, and H-BM-MSCs-Exo had a more obvious effect. In addition, N-BM-MSCs-Exo and H-BM-MSCs-Exo significantly reduced lactate dehydrogenase (LDH) release and the IL-18 levels in cell culture medium in N2a cells and primary cortical neurons. Once again H-BM-MSCs-Exo induced these effects more potently than N-BM-MSCs-Exo. All of these results demonstrated that N-BM-MSCs-Exo and H-BM-MSCs-Exo have significant neuroprotective effects against NLRP3 inflammasome-mediated pyroptosis. H-BM-MSCs-Exo has a more pronounced protective effect than N-BM-MSCs-Exo and may be used to ameliorate the progression of cerebral ischemia and hypoxia injury in patients.
外泌体在脑缺血性疾病中具有治疗潜力。在这项研究中，我们研究了常氧和低氧骨髓间充质基质细胞衍生的外泌体（N-BM-MSCs-Exo 和 H-BM-MSCs-Exo）对氧葡萄糖剥夺（OGD）损伤的神经保护作用，在小鼠神经母细胞瘤 N2a 细胞和大鼠原代皮质神经元中。OGD 损伤后 N2a 和原代皮质神经元中死亡细胞的比例显著增加，N-BM-MSCs-Exo（40μg/ml）可降低该比例，值得注意的是，H-BM-MSCs-Exo（40μg/ml）的保护作用更强。Western blot 分析表明，N-BM-MSCs-Exo 降低了 N2a 细胞中 NLRP3、ASC、Caspase-1、GSDMD-N、cleaved IL-1β 和 IL-18 的表达。然而，与 N-BM-MSCs-Exo 相比，H-BM-MSCs-Exo（40μg/ml）在抑制这些蛋白表达方面更有效。在原代皮质神经元中也得到了类似的结果。免疫荧光分析表明，N-BM-MSCs-Exo 和 H-BM-MSCs-Exo 处理后，OGD 损伤后 N2a 细胞和原代皮质神经元中 NLRP3、ASC、Caspase-1 和 GSDMD 从核到细胞质和膜的易位的共定位减少，H-BM-MSCs-Exo 的作用更为明显。此外，N-BM-MSCs-Exo 和 H-BM-MSCs-Exo 显著降低了 N2a 细胞和原代皮质神经元培养上清液中的乳酸脱氢酶（LDH）释放和 IL-18 水平。同样，H-BM-MSCs-Exo 诱导这些作用的效力强于 N-BM-MSCs-Exo。所有这些结果表明，N-BM-MSCs-Exo 和 H-BM-MSCs-Exo 对 NLRP3 炎性小体介导的细胞焦亡具有显著的神经保护作用。H-BM-MSCs-Exo 的保护作用比 N-BM-MSCs-Exo 更明显，可用于改善患者脑缺血缺氧损伤的进展。
