近期我们研究发现17β-雌二醇（βE2）在视网膜神经细胞中能激活PI3K信号通路影响细胞内Ca2+浓度（[Ca2+]i），调控重要的转录因子Nrf2的表达与活化，发挥抗氧化抗凋亡的神经保护作用，但其分子机制有待阐明。本项目拟在前期工作的基础上，以视网膜作为研究对象，通过抑制剂阻断实验、RNAi、基因过表达、ChIP及启动子生物信息学等先进的研究手段，结合FACS、Confocal、EMSA、Real-time PCR、Western Blot、免疫组化及凋亡鉴定等先进的检测技术，研究PI3K经哪些通路介导βE2调控[Ca2+]i；Ca2+信号又如何影响Nrf2的活化；激活的Nrf2/ARE通路上调了哪些靶基因的表达、最终发挥抗氧化抗凋亡保护视网膜的作用。阐明Ca2+和Nrf2/ARE信号通路介导雌激素抗氧化抗凋亡保护视网膜神经细胞的分子机制，将为寻求视网膜退行性疾病的防治措施提供新的策略。

Our previous study has found that 17β-Estradiol (βE2) has a neuroprotective effect against oxidative stress/apoptosis on retinal nerve cells by increasing the concentration of intracellular Ca2+ ([Ca2+]i) and regualting the expression/activation of an important transcription factor-Nrf2 through PI3K pathway. However, the underlying molecular mechanism is still largely unknown. Based on the previous work, these following three subjects will be investigated in this present project: which channels or pathways are involved in the changes of [Ca2+]i regulated by βE2 through PI3K; how does Ca2+ affect the activation of Nrf2, and which target genes are up-regulated by the activated Nrf2/ARE pathway, by which to exert the antioxidative/anti-apoptotic protective effects in retina. In order to accomplish those objects mentioned above, several advanced approaches including inhibitor blocking, RNAi, gene-overexpression, ChIP and promoter bioinformatics will be applied, and some detection techniques including FACS, Confocal, EMSA, Real-time PCR, Western Blot and immunohistochemistry etc. will be used in the project. Elucidating the molecular mechanism of antioxidative/anti-apoptotic protective effect of βE2 on retinal nerve cells mediated by intracelluar Ca2+ and Nrf2/ARE pathway will provide new strategy for the prevention and cure of retinal degenerative diseases.