LncRNA编码小肽NORHA-SP通过FoxO1诱导猪卵巢颗粒细胞早期凋亡的机制

卵泡闭锁是母猪繁殖力的限制因素。申请人前期鉴定了一个猪卵泡闭锁启动和颗粒细胞早期凋亡相关的lncRNA NORHA，证实其可编码一个小肽NORHA-SP。重要的是，NORHA-SP RNA和蛋白可协同诱导猪卵巢颗粒细胞早期凋亡，但机制尚不清楚。结合生物信息学分析、预试验结果和文献，推测猪卵巢颗粒细胞早期凋亡调节因子FoxO1可能是NORHA-SP RNA和蛋白的共同靶标。基于以上假设，本项目拟采用RIP等技术分析NORHA-SP RNA通过miR-183-96-182簇调控靶基因FoxO1表达的机制；采用Co-IP等技术分析NORHA-SP蛋白通过FBX4调控FoxO1蛋白稳定性的机制；采用FACS等技术分析NORHA-SP RNA和蛋白通过FoxO1对颗粒细胞早期凋亡的协同调控作用。研究结果对解析卵泡闭锁的启动机制，筛选调控母猪繁殖的小分子调节物和提高母猪繁殖力具有理论和实践意义。

Follicular atresia is a limiting factor for sow fertility. We previously identified an lncRNA NORHA associated with the initiation of porcine follicular atresia and early apoptosis of granulosa cells, and demonstrated that it encodes a small peptide, NORHA-SP. Importantly, NORHA-SP RNA and protein could synergistically induce early apoptosis of granulosa cells. However, the corresponding mechanism is still unclear. According to the bioinformatic analysis and current literature combined with our previous experiments, it is speculated that FoxO1, a regulator for early apoptosis of porcine granulosa cells, is a common target of NORHA-SP RNA and protein. Based on the above assumptions, this project intends to investigate the regulatory mechanism of NORHA-SP RNA for its target gene FoxO1 expression through the miR-183-96-182 cluster by using RIP and other technologies; to analyze the mechanism which by NORHA-SP protein regualtion of FoxO1 protein stability through FBX4 by using Co-IP and other technologies; to analyze the synergistic regulation of NORHA-SP RNA and protein on early apoptosis of granulosa cells through FoxO1 by FACS and other technologies. The results have theoretical and practical significance for exploring the molecular mechanism of follicular atresia, for screening small molecule regulators that regulate sow reproduction, and for improving sow fertility.