Department of General Medicine, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, 401120, China.
Department of Neurology, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, 401120, China.
Cardiovasc Toxicol. 2024 Mar;24(3):280-290. doi: 10.1007/s12012-024-09831-y. Epub 2024 Feb 20.
In our previous studies, the results have revealed that circRNA_102046 is significantly upregulated in plasma of patients with ischemic stroke, which closely related to NIHSS score. Human neural stem cells (hNSCs) were used for characterization and subcellular localization of circRNA_102046, and hNSCs OGD/R model was generated. The proliferation of cells was examined by CCK-8 assay. The expression levels of associated molecules were evaluated using RT-qPCR, immunofluorescence staining or western blotting. The binding and co-localization of associated molecules were also evaluated by RIP and FISH assay. Furthermore, MCAO mouse model was established to examine the effects of circRNA_102046 on the progression of ischemic stroke. Expression of circRNA_102046 was detected in the cytoplasma of hNSCs. Then OGD/R cell model was established, where the levels of circRNA_102046 was significantly up-regulated. Furthermore, knockdown of circRNA_102046 was able to enhance the proliferation and differentiation of OGD/R hNSCs. In further downstream molecular studies, the results indicated that circRNA_102046 could participate in the occurrence and development of ischemic stroke through targeting miR-493-5p. In addition, ROCK1 was identified as the putative target of miR-493-5p, and circRNA_102046 regulates the proliferation and differentiation of hNSCs via the miR-493-5p/ROCK1 signaling. More importantly, the infarct volumes of MCAO mice were remarkably reduced after the treatment with sh-circRNA_102046, which also up- and down-regulate the expression of miR-493-5p and ROCK1, respectively. Elucidating this novel pathway provides a theoretical basis for the development of new diagnostic approach and targeted treatment for ischemic stroke.
在我们之前的研究中，结果表明circRNA_102046 在缺血性中风患者的血浆中显著上调，与 NIHSS 评分密切相关。我们使用人类神经干细胞 (hNSCs) 对 circRNA_102046 进行了特征描述和亚细胞定位，并生成了 hNSCs OGD/R 模型。通过 CCK-8 测定法检查细胞的增殖。使用 RT-qPCR、免疫荧光染色或 Western blot 评估相关分子的表达水平。通过 RIP 和 FISH 测定评估相关分子的结合和共定位。此外，建立 MCAO 小鼠模型以研究 circRNA_102046 对缺血性中风进展的影响。检测 hNSCs 细胞质中 circRNA_102046 的表达。然后建立 OGD/R 细胞模型，其中 circRNA_102046 的水平显著上调。此外，circRNA_102046 的敲低能够增强 OGD/R hNSCs 的增殖和分化。在进一步的下游分子研究中，结果表明 circRNA_102046 可以通过靶向 miR-493-5p 参与缺血性中风的发生和发展。此外，ROCK1 被鉴定为 miR-493-5p 的假定靶标，circRNA_102046 通过 miR-493-5p/ROCK1 信号调节 hNSCs 的增殖和分化。更重要的是，MCAO 小鼠经 sh-circRNA_102046 治疗后，梗死体积显著减少，miR-493-5p 和 ROCK1 的表达也分别上调和下调。阐明这条新途径为缺血性中风的新诊断方法和靶向治疗的发展提供了理论基础。
