高原缺氧致中枢NFAT5异常介导HIBD后癫痫发作的机制研究

孕前移居高原子代易受缺氧打击,围产期缺氧缺血性脑损伤(Hypoxia/Ischemia Brain Damage, HIBD)可致患儿远期癫痫但机制不明。控制中枢炎症及BBB/脑膜淋巴管破坏可能是防治关键。核转录因子NFAT5是中枢炎症关键调控因子,在小胶质细胞既可被上游炎症信号激活又可调控下游炎性因子释放;还可在转录水平介导BBB/脑膜淋巴管损伤,但NFAT5在HIBD致癫痫中作用机制未明。我们前期发现NFAT5随HIBD病程动态变化并介导远期癫痫高度易感。本项目拟通过敲低或过表达NFAT5孕前移居高原大鼠新生子代缺氧模型结合组织和细胞培养,寻找HIBD后NFAT5转录调控靶向基因及启动子序列结合位点;探讨NFAT5激活介导BBB/脑膜淋巴管损伤的关键蛋白,阐明其增加神经元兴奋性免疫炎症新通路;从转录水平探索其介导HIBD后癫痫发生新机制,为预防及治疗高原低氧所致癫痫提供理论及实验依据。

The offspring of those who migrated to the high altitude before pregnancy are vulnerable to hypoxia. Hypoxia / Ischemic Brain Damage (HIBD) can cause severe sequelae such as epilepsy，cerebral palsy and mental retardation, but the reason is complex and the mechanism is still unknown. Control of central inflammation and BBB/meningeal lymphatics destruction may be the key to prevent epilepsy after hypoxia. Nuclear transcription factor (NFAT5) is proved to be a key regulatory factor in central inflammation. In microglia, NFAT5 is activated by upstream inflammatory signal pathway and also regulates the release of downstream inflammatory factors. It also mediate the injury of BBB/meningeal lymphatics at the transcription level. However, the mechanism of NFAT5 in HIBD induced-epilepsy is rarely studied. Our previous work showed that expression of NFAT5 changed dynamically with the course of HIBD and mediated the high susceptibility of long-term epilepsy. This project intends to explore the binding sites of NFAT5 transcriptional regulation target gene and promoter sequence in microglia after HIBD by knockdown or overexpression of NFAT5 in the newborn offspring animal hypoxia model of high altitude migration before pregnancy and combined with tissue and cell culture studies. To investigate the key protein of NFAT5 activation mediating BBB / meningeal lymphatics injury, and to clarify elucidate its new pathway of immune inflammation to increase neuronal excitability; To explore the mechanism of HIBD following epilepsy mediated by abnormal NFAT5 from transcriptional level, and to provide theoretical and experimental basis for the prevention and treatment of HIBD induced by high altitude hypoxia.