癫痫持续状态下lncRNA-ZNF883调控miR-181a/b-MOAP1-Bax通路促进凋亡的机制研究

Status epilepticus (SE) is a kind of acute and dangerous disease of the nervous system, with rather high disability and mortality rates, Studies have shown that non-coding RNA plays an important role in the regulation of neuronal cell apoptosis and the incidence and development of epilepsy. In the early stage, the applicant found expression difference of miR181a/b in SE using gene chip, which.was involved in the regulation of neuronal apoptosis after SE. And lncRNA-ZNF883 was predicted by informatics as lncRNAs that might bind to miR181a/b and was highly expressed in SE.Therefore, the research group proposed the following scientific idea: lncRNA-ZNF883 regulates miR181a/b-MOAP1-Bax signaling pathway,leading to neuronal apoptosis, which is regulated chronic and recurrent seizures after SE. With the epilepsy cell model and SE rat model as the intended study objects, this project observed the effects of interference on lncRNA-ZNF883 and miR-181a/b expression on MOAP1-Bax signaling pathway-related gene expression levels and neuronal apoptosis, thus to provide theoretical and experimental basis for the incidence and development mechanism of epilepsy, the screening of epilepsy prognostic markers and its clinical treatment.

癫痫持续状态（SE)为致残率和病死率较高的神经系统急危重症，研究显示非编码RNA在调控神经元细胞凋亡及癫痫发生发展中有重要作用。申请人前期利用基因芯片发现miR181a/b在SE中表达差异，参与SE后神经元凋亡的调控；并通过信息学预测lncRNA-ZNF883为miR181a/b可能结合的lncRNAs，且在SE中高表达。故课题组提出以下科学设想：lncRNA-ZNF883调控miR181a/b-MOAP1-Bax信号通路导致神经元凋亡，从而调控SE后慢性、反复痫性发作。本项目拟以癫痫细胞模型、SE大鼠模型为研究对象，观察干扰lncRNA-ZNF883、miR-181a/b表达对MOAP1-Bax信号通路相关基因表达水平及神经元凋亡的影响，为癫痫的发生发展机制、癫痫预后标志物的筛选以及临床治疗提供理论基础和实验依据。

癫痫持续状态（SE)为致残率和病死率较高的神经系统急危重症，研究显示非编码RNA在调控神经元细胞凋亡及癫痫发生发展中有重要作用。本研究构建了癫痫持续状态 SD 大鼠模型及癫痫神经元细胞模型，采用qRT-PCR法、免疫荧光法和免疫blot法检测海马神经元中基因和蛋白的表达，并采用TUNEL染色检测细胞凋亡情况。采用双荧光素酶报告法验证了miR-181b和RASSF1A之间的关系。我们论证了miR-181b模拟物显著抑制了无镁诱导的细胞凋亡，miR-181b通过直接靶向RASSF1A和激活PI3K/Akt信号通路来抑制EP的进展。同时我们发现miR-181b的上调通过调控ZNF883显著抑制了癫痫持续状态的进展。. 基于以上结果，项目在研究计划的基础上新增了对lncRNA ZNF883调控癫痫中NLRP3免疫酶体激活机制的研究。分别用匹罗卡品和无镁海马神经元细胞建立大鼠和细胞EP模型，研究ZNF883、miR- 138-5p、泛素特异性肽酶47（USP47）和NLRP3的双向表达及可能作用机制。研究发现lncRNA ZNF883的过表达可下调miR-138-5p的表达，而miR-138-5p的上调可抑制USP47的表达。我们的深入探索进一步发现，抑制miR-138-5p在一定程度上抑制了lncRNA ZNF883沉默对ep相关神经元凋亡、神经炎症和癫痫样放电的抑制作用。为癫痫的发生发展机制、癫痫预后标志物的筛选以及临床治疗提供理论基础和实验依据。