Department of Neurosurgery, Haikou Affiliated Hospital of Central South University Xiangya School of Medicine, No. 43, Renmin Avenue, Meilan District, Haikou, 570208, Hainan Province, People's Republic of China.
Neurochem Res. 2023 Jun;48(6):1811-1821. doi: 10.1007/s11064-023-03866-3. Epub 2023 Jan 30.
Ischemic stroke (IS) is the most common type of stroke and the second leading cause of death overall. Neural stem cells play protective roles in IS, but the underlying mechanism remains to be determined. Neural stem cells (NSC) were obtained from the fetal brain tissue of C57BL/6J mice. NSC-derived exosomes (NSC-Exos) were identified in the conditioned medium. Internalization of NSC-Exos was analyzed by fluorescence microscopy. In vitro microglia ischemic stroke injury model was induced using oxygen glucose deprivation/re-oxygenation (OGD/R) method. Cell viability and inflammation were analyzed by MTT, qPCR, ELISA and Western blotting assay. Interaction between ZEB1 and the promoter of GPR30 was verified by luciferase assay and chromatin immunoprecipitation. NSC-Exos prevented OGD/R-mediated inhibition of cell survival and the production of inflammatory cytokines in microglia cells. NSC-Exos increased ZEB1 expression in OGD/R-treated microglia. Down-regulation of ZEB1 expression in NSC-Exos abolished NSC-Exos' protective effects on OGD/R-treated microglia. ZEB1 bound to the promoter region of GPR30 and promoted its expression. Inhibiting GPR30 reversed NSC-Exos effects on cell viability and inflammation injury in OGD/R-treated microglia. Our study demonstrated that NSC exerted cytoprotective roles through release of exosomal ZEB1,which transcriptionally upregulated GPR30 expression, resulting in a reduction in TLR4/NF-κB pathway-induced inflammation. These findings shed light on NSC-Exos' cytoprotective mechanism and highlighted its potential application in the treatment of IS.
缺血性脑卒中（IS）是最常见的脑卒中类型，也是总体上的第二大致死原因。神经干细胞在 IS 中发挥保护作用，但潜在机制仍有待确定。神经干细胞（NSC）从 C57BL/6J 小鼠的胎脑组织中获得。在条件培养基中鉴定出 NSC 衍生的外泌体（NSC-Exos）。通过荧光显微镜分析 NSC-Exos 的内化。使用氧葡萄糖剥夺/复氧（OGD/R）方法诱导体外小胶质细胞缺血性脑卒中损伤模型。通过 MTT、qPCR、ELISA 和 Western blot 分析细胞活力和炎症。通过荧光素酶测定和染色质免疫沉淀验证 ZEB1 与 GPR30 启动子之间的相互作用。NSC-Exos 可防止 OGD/R 介导的小胶质细胞存活抑制和炎症细胞因子的产生。NSC-Exos 增加了 OGD/R 处理的小胶质细胞中 ZEB1 的表达。下调 NSC-Exos 中的 ZEB1 表达消除了 NSC-Exos 对 OGD/R 处理的小胶质细胞的保护作用。ZEB1 结合到 GPR30 的启动子区域并促进其表达。抑制 GPR30 逆转了 NSC-Exos 对 OGD/R 处理的小胶质细胞中细胞活力和炎症损伤的作用。我们的研究表明，NSC 通过释放外泌体 ZEB1 发挥细胞保护作用，转录上调 GPR30 表达，从而减少 TLR4/NF-κB 通路诱导的炎症。这些发现揭示了 NSC-Exos 的细胞保护机制，并强调了其在 IS 治疗中的潜在应用。
