Zhang Qian, Yi Yan, Chen Tiange, Ai Ying, Chen Ziyang, Liu Ganzhi, Tang Zexuan, Chen Jianwei, Xu Tao, Chen Xin, Liu Jinfang, Xia Yuguo
Department of Neurosurgery, Xiangya Hospital Central South University, Changsha, Hunan, CN, 410008, China.
National Clinical Research Center for Geriatric Disorders, Xiangya Hospital Central South University, Changsha, Hunan, CN, 410008, China.
J Nanobiotechnology. 2025 Apr 23;23(1):311. doi: 10.1186/s12951-025-03390-2.
Endogenous neurogenesis could promote stroke recovery. Furthermore, anti-inflammatory phenotypical microglia (M2-microglia) could facilitate Neural Stem Cell (NSC)-mediated neurogenesis following Ischemic Stroke (IS). Nonetheless, the mechanisms through which M2 microglia influence NSC-mediated neurogenesis post-IS remain unclear. On the other hand, M2 microglia-derived small Extracellular Vesicles (M2-sEVs) could exert phenomenal biological effects and play significant roles in cell-to-cell interactions, highlighting their potential involvement in NSC-mediated neurogenesis post-IS, forming the basis of this study.
M2-sEVs were first isolated from IL-4-stimulated microglia. For in vivo tests, M2-sEVs were intravenously injected into mice every day for 14 days after transient Middle Cerebral Artery Occlusion (tMCAO). Following that, the infarct volume and neurological function, as well as NSC proliferation in the Subventricular Zone and dentate gyrus, migration, and differentiation in the infarct area, were examined. For in vitro tests, M2-sEVs were administered to NSC subjected to Oxygen-Glucose Deprivation (OGD) and then reoxygenation, after which NSC proliferation and differentiation were assessed. Finally, M2-sEVs were subjected to microRNA sequencing to explore the regulatory mechanisms.
Our findings revealed that M2-sEVs reduced the infarct volume and increased the neurological score in mice post-tMCAO. Furthermore, M2-sEV treatment promoted NSC proliferation and neuronal differentiation both in vivo and in vitro. Additionally, microRNA sequencing revealed miR-93-5p and miR-25-3p enrichment in M2-sEVs. Inhibitors of these miRNAs prevented TGFBR, PTEN, and FOXO3 downregulation in NSC, reversing M2-sEVs' beneficial effects on neurogenesis and sensorimotor recovery.
M2-sEVs increased NSC proliferation and neuronal differentiation, and protected against IS, at least partially, via delivering miR-25-3p and miR-93-5p to downregulate TGFBR, PTEN, and FOXO3 expression in NSC.
内源性神经发生可促进中风恢复。此外，抗炎表型的小胶质细胞（M2小胶质细胞）可促进缺血性中风（IS）后神经干细胞（NSC）介导的神经发生。然而，M2小胶质细胞影响IS后NSC介导的神经发生的机制仍不清楚。另一方面，M2小胶质细胞衍生的小细胞外囊泡（M2-sEVs）可发挥显著的生物学效应，并在细胞间相互作用中发挥重要作用，这突出了它们可能参与IS后NSC介导的神经发生，构成了本研究的基础。
首先从白细胞介素-4刺激的小胶质细胞中分离出M2-sEVs。对于体内试验，在短暂性大脑中动脉闭塞（tMCAO）后每天给小鼠静脉注射M2-sEVs，持续14天。之后，检测梗死体积和神经功能，以及脑室下区和齿状回中NSC的增殖、梗死区域中NSC的迁移和分化。对于体外试验，将M2-sEVs给予经历氧糖剥夺（OGD）然后再复氧的NSC，之后评估NSC的增殖和分化。最后，对M2-sEVs进行微小RNA测序以探索调控机制。
我们的研究结果显示，M2-sEVs可减小tMCAO后小鼠的梗死体积并提高神经评分。此外，M2-sEVs治疗在体内和体外均促进了NSC的增殖和神经元分化。此外，微小RNA测序显示M2-sEVs中富含miR-93-5p和miR-25-3p。这些微小RNA的抑制剂可阻止NSC中转化生长因子β受体（TGFBR）、第10号染色体缺失的磷酸酶和张力蛋白同源物（PTEN）以及叉头框蛋白O3（FOXO3）的下调，逆转M2-sEVs对神经发生和感觉运动恢复的有益作用。
M2-sEVs可增加NSC的增殖和神经元分化，并至少部分地通过递送miR-25-3p和miR-93-5p来下调NSC中TGFBR、PTEN和FOXO3的表达，从而对IS起到保护作用。
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