Department of Neurology, Lianyungang Hospital affiliated to Xuzhou Medical College, Lianyungang, Jiangsu Province, China.
Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Cell Cycle. 2023 Feb;22(4):403-418. doi: 10.1080/15384101.2022.2121358. Epub 2022 Dec 22.
MicroRNA-141-3p (miR-141-3p) has been found to be altered in the brain following a stroke. Herein, we investigate the impact of miR-141-3p on the apoptosis of neural stem cells (NSCs) in mice with middle cerebral artery occlusion (MCAO) and the potential mechanisms involved. Eight-week-old mice were injected intracerebroventricularly with miR-141-3p, antagomir-141-3p, or agomir negative control 2 h before MCAO, and animal behavior tests and infraction volume measurements were performed 24 h later. MCAO-mediated brain injury and NSCs apoptosis were observed by H&E, TTC, and TUNEL staining. The expression of cleaved caspase-3 and Ki67 was detected by western blotting. The luciferase reporter assay proved that miR-141-3p in combination with its target gene PBX homeobox 1 (PBX1). Exogenous miR-141-3p (agomir-141-3p) treatment increased infraction volume and brain edema and damaged neurological functions compared to control mice. Agomir-141-3p increased miR-141-3p expression in brain tissue of mice with MCAO and suppressed PBX1 expression. The effects of the agomir-141-3p-induced apoptosis in NSCs treated with oxygen-glucose deprivation (OGD)/reoxygenation (R) were abolished by PBX1 overexpression. The results from UCSC and JASPAR database showed that prokineticin 2 (PROK2) was a transcription factor of PBX1. The expression of PROK2 was transcriptionally regulated by PBX1 using RT-PCR and western blot assays. The effects of the apoptosis-promoting caused by PBX1 inhibition in NSCs treated with OGD/R were reversed by PROK2 inhibition. In conclusion, the miR-141-3p/PBX1/PROK2 axis might be a novel therapeutic target for the apoptosis of NSCs in MCAO.
miR-141-3p（miR-141-3p）在中风后大脑中发生改变。在此，我们研究了 miR-141-3p 对大脑中动脉闭塞（MCAO）小鼠神经干细胞（NSC）凋亡的影响及其潜在机制。8 周龄的小鼠在 MCAO 前 2 小时经侧脑室注射 miR-141-3p、antagomir-141-3p 或 agomir 阴性对照物，24 小时后进行动物行为测试和梗死体积测量。通过 H&E、TTC 和 TUNEL 染色观察 MCAO 介导的脑损伤和 NSCs 凋亡。通过 Western blot 检测裂解的 caspase-3 和 Ki67 的表达。荧光素酶报告基因检测证明 miR-141-3p 与其靶基因 PBX 同源盒 1（PBX1）结合。与对照小鼠相比，外源性 miR-141-3p（agomir-141-3p）处理增加了梗死体积和脑水肿，并损害了神经功能。Agomir-141-3p 增加了 MCAO 小鼠脑组织中 miR-141-3p 的表达，并抑制了 PBX1 的表达。在 OGD/R 处理的 NSCs 中，过表达 PBX1 可消除 agomir-141-3p 诱导的细胞凋亡。UCSC 和 JASPAR 数据库的结果表明，促动力素 2（PROK2）是 PBX1 的转录因子。使用 RT-PCR 和 Western blot 测定，PROK2 转录调控 PBX1 的表达。在 OGD/R 处理的 NSCs 中，抑制 PBX1 引起的凋亡作用被 PROK2 抑制所逆转。综上所述，miR-141-3p/PBX1/PROK2 轴可能是 MCAO 中 NSCs 凋亡的新治疗靶点。
