Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, 430060, China.
J Mol Histol. 2024 Oct;55(5):699-707. doi: 10.1007/s10735-024-10224-2. Epub 2024 Jul 17.
Long non-coding RNAs (LncRNAs) play a substantial role in the process of cerebral ischemia-reperfusion injury (CIRI). The present work aimed to determine the probable mechanism by which LncRNA TUG1 exacerbates CIRI via the miR-340-5p/phosphatase and tensin homolog (PTEN) pathway. After developing a middle cerebral artery occlusion/reperfusion (MCAO/R) model, pcDNA-TUG1 together with miR-340-5p agomir were administrated in vivo. Furthermore, the neurologic defects in rats were assessed by a modified neurological severity score. Moreover, 2,3,5-Triphenyl-2 H-tetrazolium chloride stain-step was performed to determine the brain's infarct size. In addition, western blotting, immunohistochemistry, and qRT-PCR experiments were utilized for gauging the proteomic/genomic expression-profiles. Luciferase reporter assay validated correlations across TUG1, miR-340-5p, together with PTEN. The results indicated relatively reduced miR-340-5p levels in MCAO/R models, while upregulated TUG1 levels. The pcDNA-TUG1-treated rats indicated increasing neurological dysfunction, whereas the miR-340-5p agomir-treated rats showed improvement. Furthermore, miR-340-5p was determined to be the expected and confirmed TUG1 target. All things considered, the findings suggested that PTEN can serve as the target of miR-340-5p. In addition, TUG1 served as a miR-340-5p ceRNA, which promotes PTEN modulation. Furthermore, TUG1 overexpression decreased miR-340-5p's capacity to fend against CIRI. Conclusively, this work proved that in CIRI, targeting the TUG1/miR-340-5p/PTEN regulatory axis is a viable approach for the treatment of ischemic stroke.
长链非编码 RNA（LncRNA）在脑缺血再灌注损伤（CIRI）过程中发挥重要作用。本研究旨在通过 LncRNA TUG1 通过 miR-340-5p/磷酸酶和张力蛋白同源物（PTEN）通路加重 CIRI 的可能机制。建立大脑中动脉闭塞/再灌注（MCAO/R）模型后，体内给予 pcDNA-TUG1 与 miR-340-5p agomir。此外，通过改良神经严重程度评分评估大鼠的神经缺陷。此外，通过 2,3,5-三苯基-2 H-四唑氯化物染色步骤确定脑梗死面积。此外，还进行了 Western blot、免疫组化和 qRT-PCR 实验以评估蛋白质组/基因组表达谱。荧光素酶报告基因实验验证了 TUG1、miR-340-5p 与 PTEN 之间的相关性。结果表明，MCAO/R 模型中 miR-340-5p 水平相对降低，而 TUG1 水平升高。pcDNA-TUG1 处理的大鼠表现出神经功能障碍增加，而 miR-340-5p agomir 处理的大鼠则有所改善。此外，miR-340-5p 被确定为预期和证实的 TUG1 靶标。综上所述，这些发现表明 PTEN 可以作为 miR-340-5p 的靶点。此外，TUG1 作为 miR-340-5p 的 ceRNA，促进了 PTEN 的调节。此外，TUG1 的过表达降低了 miR-340-5p 对抗 CIRI 的能力。总之，这项工作证明，在 CIRI 中，靶向 TUG1/miR-340-5p/PTEN 调控轴是治疗缺血性中风的一种可行方法。
