lncRNAGRN-miR-6615-5p调控TGF-β1/SMAD7信号通路参与氨气诱导的鸡胸腺细胞凋亡机理

集约化鸡舍中的氨气是危害肉鸡最大的有毒气体，胸腺是免疫应激反应研究的主要对象，然而鸡氨气中毒导致胸腺损伤的机理研究甚少。前期发现鸡氨气中毒影响了胸腺组织lncRNAGRN、miR-6615-5p以及SMAD7表达，生物信息学预测存在靶向关系。故本项目拟在氨暴露模型和胸腺细胞体外氨暴露模型基础上，采用荧光素酶报告基因、基因过表达和敲低技术，验证lncRNAGRN、miR-6615-5p及SMAD7的靶向关系，应用透射电镜、流式细胞术、TUNEL法、实时定量荧光PCR和免疫印迹技术，通过检测TGF-β1/SMAD7信号通路和细胞凋亡相关基因的表达、细胞凋亡等，阐明lncRNAGRN-miR-6615-5p通过调节TGF-β1/SMAD7信号通路在氨气暴露导致的肉鸡胸腺细胞凋亡中的作用机理。为防治鸡氨气中毒提供新的思路和理论依据，为集约化家禽生产提供参考。

Ammonia (NH3) in intensive chicken house is the most harmful poisonous gas to broilers. Thymus is the main research object on stress response. However, little research has been done on the mechanism of thymus injury caused by NH3 poisoning in chickens. In previous studies, we found that chicken NH3 poisoning affected the expression of lncRNAGRN, miR-6615-5p, and SMAD7; Bioinformatics Prediction that there was a targeting relationship. Therefore, on the basis of ammonia exposure model and in vitro ammonia exposure model of thymocytes, the project wants to verify the targeting relationship of lncRNAGRN, miR-6615-5p, and SMAD7 by using luciferase reporter gene, gene overexpression and knock-down techniques, and to detect TGF-β1/SMAD7 signaling pathway and cell apoptosis by using transmission electron microscopy, flow cytometry, TUNEL, real-time quantitative PCR, and immunoblotting techniques. To elucidate the mechanism of lncRNAGRN-miR-6615-5p in thymocyte apoptosis induced by NH3 exposure in broilers by regulating TGF-β1/SMAD7 signaling pathway. To provide new ideas and theoretical basis for the prevention and treatment of NH3 poisoning in chickens, and to provide reference for intensive poultry production.