Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu Province, P.R. China.
College of Health Sciences, Jiangsu Normal University, Xuzhou, P.R., China.
J Cell Physiol. 2018 Sep;233(9):7022-7034. doi: 10.1002/jcp.26498. Epub 2018 Mar 25.
Epilepsy is a group of neurological disorders characterized by epileptic seizures. In this study, we aim to explore the role of microRNA-421 (miR-421) in hippocampal neurons of epilepsy mice via the TLR/MYD88 pathway. Forty mice were randomly served as the normal and model (established as epilepsy model) groups. Hippocampal neurons were assigned into seven groups with different transfections. The RT-qPCR and western blotting were conducted to examine the expression of miR-421 TLR2, TLR4, MYD88, Bax, Bcl-2, p53, Beclin-1, and LC3II/LC3I. Cell proliferation and apoptosis were detected by MTT and flow cytometry.MYD88 is a target gene of miR-421. Model mice showed elevated expression of TLR2, TLR4, MYD88, Bax, p53, Beclin-1, and LC3II/LC3I but reduced expression of miR-421 and Bcl-2. In vitro experiments reveals that overexpression of miR-421 inhibited the TLR/MYD88 pathway. Besides, overexpressed miR-421 declined cell apoptosis but increased cell proliferation. It reveals that miR-421 targeting MYD88 could inhibit the apoptosis and autophagy of hippocampal neurons in epilepsy mice by down-regulating the TLR/MYD88 pathway.
癫痫是一组以癫痫发作为特征的神经障碍。在这项研究中,我们旨在通过 TLR/MYD88 通路探索 microRNA-421 (miR-421) 在癫痫小鼠海马神经元中的作用。将 40 只小鼠随机分为正常组和模型组(建立癫痫模型)。将海马神经元分为 7 组进行不同的转染。通过 RT-qPCR 和 Western blot 检测 miR-421、TLR2、TLR4、MYD88、Bax、Bcl-2、p53、Beclin-1 和 LC3II/LC3I 的表达。通过 MTT 和流式细胞术检测细胞增殖和凋亡。MYD88 是 miR-421 的靶基因。模型小鼠 TLR2、TLR4、MYD88、Bax、p53、Beclin-1 和 LC3II/LC3I 的表达升高,miR-421 和 Bcl-2 的表达降低。体外实验表明,miR-421 的过表达抑制了 TLR/MYD88 通路。此外,过表达 miR-421 可降低细胞凋亡,增加细胞增殖。这表明 miR-421 通过靶向 MYD88 抑制 TLR/MYD88 通路,可抑制癫痫小鼠海马神经元的凋亡和自噬。