Department of Rehabilitation Medicine, Qujing No.1 Hospital, Qujing, 655000, Yunnan, China.
Cell Biochem Biophys. 2024 Dec;82(4):3629-3643. doi: 10.1007/s12013-024-01452-0. Epub 2024 Jul 31.
To investigate the effect of miR-3571 on traumatic brain injury (TBI) via the regulation of neuronal apoptosis through F-box-only protein 31/phosphoinositide 3-kinase/protein kinase B (Fbxo31/PI3K/AKT). We established TBI rat and cell models. Hematoxylin‒eosin (HE) and Nissl staining were used to observe brain injury and the number of Nissl bodies, respectively. Cell proliferation and apoptosis were assessed by 5-ethynyl-2'-deoxyuridine (EdU), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and flow cytometry. Gene and protein expression was measured via reverse transcription quantitative polymerase chain reaction (RT‒qPCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). In this study, miR-3571 was highly expressed in TBI models. Inhibition of miR-3571 expression can suppress autophagy, reduce the expression of proinflammatory cytokines, and reduce neuronal apoptosis, thus alleviating the pathological conditions of tissue congestion, edema and structural damage after TBI. These experiments demonstrated that miR-3571 could target and regulate the level of Fbxo31. Knockdown of Fbxo31 weakened the remission effect of the miR-3571 inhibitor on TBI and promoted neurological damage; moreover, overexpression of Fbxo31 enhanced the protective effect on neural function, whereas the PI3K/AKT pathway inhibitor LY294002 increased the damage caused by miR-3571 on neural function and weakened the protective effect of Fbxo31. In conclusion, miR-3571 regulates the PI3K/AKT signaling pathway by reducing Fbxo31 expression, promotes neuronal apoptosis and exacerbates TBI.
为了通过调节神经元凋亡来研究 miR-3571 对创伤性脑损伤(TBI)的影响,研究人员利用 F-box 仅蛋白 31/磷酸肌醇 3-激酶/蛋白激酶 B(Fbxo31/PI3K/AKT)实现这一目标。他们建立了 TBI 大鼠和细胞模型。苏木精-伊红(HE)和尼氏染色分别用于观察脑损伤和尼氏小体的数量。通过 5-乙炔基-2'-脱氧尿苷(EdU)、末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记(TUNEL)和流式细胞术评估细胞增殖和凋亡。通过逆转录定量聚合酶链反应(RT-qPCR)、Western blot 和酶联免疫吸附测定(ELISA)测量基因和蛋白表达。在这项研究中,miR-3571 在 TBI 模型中高表达。抑制 miR-3571 表达可以抑制自噬,降低促炎细胞因子的表达,减少神经元凋亡,从而减轻 TBI 后组织充血、水肿和结构损伤的病理状况。这些实验表明,miR-3571 可以靶向并调节 Fbxo31 的水平。Fbxo31 的敲低削弱了 miR-3571 抑制剂对 TBI 的缓解作用并促进了神经损伤;此外,Fbxo31 的过表达增强了对神经功能的保护作用,而 PI3K/AKT 通路抑制剂 LY294002 增加了 miR-3571 对神经功能的损伤并削弱了 Fbxo31 的保护作用。总之,miR-3571 通过降低 Fbxo31 表达来调节 PI3K/AKT 信号通路,促进神经元凋亡并加重 TBI。