【摘要】 目的 探究lncRNA H19过表达与癫痫Reelin信号通路表达变化及海马神经元迁移的相关性。方法 取新生24 h内SD大鼠海马体外培养神经元，随机分为4组，分别为空白组、癫痫组、NC组、H19组，其中癫痫组、NC组、H19组均经癫痫造模处理，且NC组经转染空载腺病毒预处理，H19组经转染lncRNA H19过表达腺病毒预处理。观察各组神经元形态学变化，并采用实时荧光定量PCR技术检测各组lncRNA H19、Reelin及其下游分子VLDLR、Dab1的mRNA表达水平及Western blot半定量和免疫荧光技术检测lncRNA H19的表达变化对各组Reelin、VLDLR、Dab1表达的影响。结果 与空白组相比，癫痫组神经元突触连接增多，网络更复杂，H19组该趋势更明显。癫痫组lncRNA H19表达升高（P=0.001）,H19组进一步升高（P<0.001）。癫痫组Reelin、Dab1、VLDLR的mRNA和蛋白表达均下降（P<0.05）,H19组各分子表达进一步下降（P<0.05）。免疫荧光显示癫痫组Reelin、Dab1荧光强度下降，H19组下降更显著（P<0.05）。结论 lncRNA H19过表达可抑制Reelin信号通路相关分子的表达，lncRNA H19可能通过抑制Reelin信号通路参与调控癫痫中神经元迁移过程。更多还原

【Abstract】 Objective To investigate the relevance of lncRNA H19 overexpression on Reelin signaling pathway expression changes and hippocampal neuronal migration in epilepsy.Methods Primary hippocampal neurons were isolated from the hippocampi of neonatal SD rats within 24 hours after birth, cultured in vitro, and then randomly divided into four groups: the blank group, the epilepsy group, the NC group, and the H19 group.The epilepsy, NC, and H19 groups were treated with epilepsy modeling, and the NC group was pretreated with transfected null-loaded adenovirus and the H19 group was pretreated with transfected lncRNA H19 overexpression adenovirus. Neuronal morphology was observed across groups. Quantitative real-time PCR was used to measure the mRNA expression levels of lncRNA H19, Reelin, and its downstream molecules VLDLR and Dab1. Western blot analysis and immunofluorescence staining were employed to determine the effects of lncRNA H19 expression changes on the protein expression of Reelin, VLDLR, and Dab1 in each group.Results Compared with the blank group, the neurons in the epilepsy group exhibited increased synaptic connections and a more complex neural network, and this tendency was more pronounced in the H19 group. The expression of lncRNA H19 was significantly elevated in the epilepsy group(P=0.001), and it was further upregulated in the H19 group(P<0.001). In the epilepsy group, both the mRNA and protein expressions of Reelin, Dab1 and VLDLR were significantly decreased(P<0.05), and their expressions were further reduced in the H19 group(P<0.05).Immunofluorescence assays revealed that the fluorescence intensities of Reelin and Dab1 were decreased in the epilepsy group, with a more significant reduction observed in the H19 group(P<0.05).Conclusion Overexpression of lncRNA H19 inhibits the expression of molecules related to the Reelin signaling pathway.lncRNA H19 may be involved in regulating neuronal migration in epilepsy by inhibiting the Reelin signaling pathway.更多还原