Department of Functional Neurosurgery, Beijing Neurosurgical Institute, Capital Medical University, Beijing, China.
Beijing Key Laboratory of Neurostimulation, Beijing, China.
Cell Prolif. 2020 Aug;53(8):e12856. doi: 10.1111/cpr.12856. Epub 2020 Jul 10.
Glial cell activation contributes to the inflammatory response and occurrence of epilepsy. Our preliminary study demonstrated that the long non-coding RNA, H19, promotes hippocampal glial cell activation during epileptogenesis. However, the precise mechanisms underlying this effect remain unclear.
H19 and let-7b were overexpressed or silenced using an adeno-associated viral vector in vivo. Their expression in a kainic acid-induced epilepsy model was evaluated by real-time quantitative PCR, fluorescence in situ hybridization, and cytoplasmic and nuclear RNA isolation. A dual-luciferase reporter assay was used to evaluate the direct binding of let-7b to its target genes and H19. Western blot, video camera monitoring and Morris water maze were performed to confirm the role of H19 and let7b on epileptogenesis.
H19 was increased in rat hippocampus neurons after status epilepticus, which might be due to epileptic seizure-induced hypoxia. Increased H19 aggravated the epileptic seizures, memory impairment and mossy fibre sprouting of the epileptic rats. H19 could competitively bind to let-7b to suppress its expression. Overexpression of let-7b inhibited hippocampal glial cell activation, inflammatory response and epileptic seizures by targeting Stat3. Moreover, overexpressed H19 reversed the inhibitory effect of let-7b on glial cell activation.
LncRNA H19 could competitively bind to let-7b to promote hippocampal glial cell activation and epileptic seizures by targeting Stat3 in a rat model of temporal lobe epilepsy.
神经胶质细胞的激活参与了炎症反应和癫痫的发生。我们的初步研究表明，长链非编码 RNA H19 在癫痫发生过程中促进海马神经胶质细胞的激活。然而，其确切机制尚不清楚。
通过腺相关病毒载体在体内过表达或沉默 H19 和 let-7b。通过实时定量 PCR、荧光原位杂交和细胞质及核 RNA 分离评估它们在海人酸诱导的癫痫模型中的表达。双荧光素酶报告基因实验用于评估 let-7b 与其靶基因和 H19 的直接结合。Western blot、摄像机监测和 Morris 水迷宫用于确认 H19 和 let7b 在癫痫发生中的作用。
在癫痫持续状态后，大鼠海马神经元中 H19 增加，这可能是由于癫痫发作引起的缺氧。增加的 H19 加重了癫痫发作、癫痫大鼠的记忆障碍和苔藓纤维发芽。H19 可以与 let-7b 竞争结合，从而抑制其表达。过表达 let-7b 通过靶向 Stat3 抑制海马神经胶质细胞的激活、炎症反应和癫痫发作。此外，过表达的 H19 逆转了 let-7b 对神经胶质细胞激活的抑制作用。
在颞叶癫痫大鼠模型中，长链非编码 RNA H19 可通过与 let-7b 竞争结合，靶向 Stat3 促进海马神经胶质细胞的激活和癫痫发作。
