电化学分析体系用于牛乳腺炎的凋亡诱导自我保护机制的研究

Mastitis is one of the common diseases in dairy cows, with the characteristics of high incidence, infection and spreading, which highly endangers the health of mammary gland in dairy cows. In the early stage of mastitis, mammary epithelial cells often die in the form of apoptosis. This type of death, apoptosis, will cause mammary epithelial cells to be swallowed by phagocytic cells, avoiding the initiation of the downstream inflammatory reactions and the deterioration of mammary gland inflammation. Electrochemical analysis technology, which has the feature of good sensitivity, high accuracy, possesses incomparable advantages compared with traditional research methods. In this project, suitable NF-κB (key NF-κB pathway activation protein) and caspase-3 (key apoptosis pathway activation protein) electrochemical analysis systems are fabricated, respectively. Then, the above systems are used to quantify the activation of NF-κB and caspase-3 in both endotoxin induced inflammation model of mammary epithelial cells and tumor necrosis factor (TNF) -α induced apoptosis model of mammary epithelial cells. By comparison the activation degree of two pathways between inflammation model and apoptosis model of mammary epithelial cells, look for the potential relationship of the two signal pathways. This project is a supplement to the related mechanism study in bovine mastitis. Induction of apoptosis of mammary inflammatory epithelial cells, avoiding the further development of inflammation, maintaining the integrity of mammary gland, has vital significance in protection the health of mammary gland in dairy cows.

乳腺炎是奶牛的常见疾病，具有高发性、易感染性和易传播性的特点，严重危害奶牛的乳腺健康。乳腺炎发病早期，乳腺上皮细胞主要以细胞凋亡的方式死亡。这种凋亡式的自我死亡方式会导致乳腺上皮细胞直接被吞噬细胞吞噬，避免了下游一系列炎症反应的引发和炎症效应的加重。电化学分析技术灵敏度好，精确度高，具有传统研究手段无法比拟的优势。本项目以电化学分析技术为主要实验手段，构建了适用于检测NF-κB与caspase-3活性的电化学分析体系，通过对内毒素诱导的乳腺上皮细胞炎症模型以及肿瘤坏死因子-α诱导的凋亡模型中NF-κB和caspase-3进行定量，比对不同模型中两条通路的激活程度，以寻找两条通路之间潜在对应关系。本项目从凋亡诱导的自我保护机制出发，提出通过增加诱导奶牛乳腺炎症上皮细胞凋亡以最大程度的保持乳腺的完整性，避免乳腺炎的交叉感染和进一步发展，对保护奶牛乳腺健康具有重要意义。

当前乳腺炎对奶牛产业的危害日益严重。本项目主要以电化学分析技术为主要研究手段，辅以其他传统方法，通过1）对生物分子修饰电极的界面行为研究，2）不同纳米材料光电性质及基于纳米载体信号放大系统的研究，3）炎症通路中关键节点蛋白NF-κB新型生物传感电化学分析体系的构建，4）凋亡通路中关键节点蛋白PARP及Caspase-3生物传感分析体系的构建，实现了炎症和凋亡通路中的关键节点蛋白的精确定量。研究为探讨牛乳腺炎发病中的炎症机制与凋亡机制之间的关系提供新的方法，为通过调控死亡方式进行牛乳腺炎症治疗以最大程度保持乳腺完整性，避免乳腺炎的交叉感染和进一步发展提供思路，对保护奶牛乳腺健康具有重要意义。