奶牛乳腺炎是严重危害奶牛业的重要疾病，奶牛乳腺上皮细胞间充质转化（EMT）是乳腺损伤的重要病理过程。NF-κB和TGF-β/Smad为介导乳腺上皮细胞间充质转化的核心信号转导通路，申请人研究证实6-溴靛玉红-3’-肟通过调控NF-κB通路抑制乳腺损伤，但6-溴靛玉红-3-肟调控NF-κB 和TGF-β/Smad介导的乳腺上皮细胞间充质转化的机理仍不清楚。本项目通过建立原代奶牛乳腺上皮细胞间充质转化模型，聚焦NF-κB 和TGF-β/Smad信号转导通路的核心环节，基于信号通路抑制、免疫印迹、流式细胞术分析、Transwell小室及跨上皮细胞电阻测定等技术，解析NF-κB 和TGF-β/Smad信号转导途径中受体、转录因子和效应分子的变化规律，为阐明6-溴靛玉红-3’-肟调控奶牛乳腺上皮细胞间充质转化的分子机制，寻找干预奶牛乳腺损伤的新型药物靶标奠定理论基础。

Dairy cow mastitis is one of the important diseases which cause serious damage to the dairy industry, and the transformation of epithelial mesenchymal transition (EMT) is the key issue to breast damage. It was reported that NF-κB and TGF-β/Smad signal pathway were the important signaling pathways for BMECs’ EMT. It was reported that 6-Br-indirubin-3'-monoxime could regulate mammary damage through NF-κB signal pathway by applicant. But how BIO regulated EMT of BMECs via NF-κB and TGF-β/Smad signal pathways were not clear. The culture system including BMECs and LPS/TGFβ1 was constructed. The EMT process was analysed through signal pathway inhibition, western blot, flow cytometry analysis, Transwell chamber migration observation and trans-epithelium electrical resistant (TEER) measurement, et al. In conclusion, the mechnism of BMECs’ EMT regulated according to NF-κB and TGF-β/Smad were studied in detail. It was a foundation to clarify the cow mammary gland damage, which provided theoretical basis for search of novel drug targets for the intervention of mammary gland damage.