牛蒡苷元对3D4/21细胞免疫功能的调节作用及机制

Searching for a safe and efficient immune-potentiator has become the focus of researchers in the field of veterinary medicine. Arctigenin (ARG), an active ingredient exist in traditional Chinese medicine Fructus Arctii, has been proven exerting an immune-regulatory effects through regulating the bio-behavior of macrophages, however, the mechanism still remains enigmatic. We are intended to take porcine alveolar macrophages as the research object. Firstly, in order to investigate the stimulation effect of ARG on 3D4/21 cells, the changes of cellular immune function including phagocytosis and cytokines secretion are going to be detected by ELISA, RT-QPCR etc.. Then, the effect of ARG on immune-associated signal pathways, such as TLRs、NF-κB、MAPKs, will be detected by Western Blot, immune-fluorescence etc.. And the target molecule thereof and the cellular signal transduction will be demonstrated. In addition, the effect of ARG on immune-regulating in piglets will be evaluated in clinical trials. This project aims to excavate the function of ARG in immune regulation and explore the underlying mechanism. Thus offers foundation for the application of ARG as an immune-potentiator in improving the immunity of animals, and promote actively the development of traditional Chinese medicine in veterinary clinical field.

寻找安全、高效的免疫增强剂已成为兽药领域的研究热点。有研究表明，中药牛蒡子的活性成分牛蒡苷元（ARG）可以通过作用于巨噬细胞来调节机体免疫功能，但其作用机制迄今尚不明确。本项目拟以猪肺泡巨噬细胞（3D4/21）为研究对象，首先利用ELISA、RT-QPCR等技术，检测ARG对3D4/21细胞吞噬及细胞因子分泌等功能的影响，以探究ARG对3D4/21细胞的活化作用。然后，采用western blotting、免疫荧光等方法检测ARG对TLRs、NF-κB、MAPKs等免疫调节相关信号通路的影响，从而寻找ARG作用的分子靶标，并揭示其诱导3D4/21细胞活化的信号转导途径。最后，结合动物试验评价ARG在仔猪体内的免疫调节效果。本项目旨在深入探索ARG的免疫调节功能，并阐释其作用机制。预期结果将为ARG提高畜禽机体免疫力的临床应用奠定理论基础，也将为推动中药在兽医临床方面的研究开发起到积极作用。

寻找安全、高效的免疫增强剂已成为兽药领域的研究热点。有研究表明，中药牛蒡子的活性成分牛蒡苷元（ARG）可以通过作用于巨噬细胞来调节机体免疫功能，但其作用机制迄今尚不明确。本项目以猪肺泡巨噬细胞（3D4/21）为研究对象，研究发现ARG对3D4/21无明显毒性作用，且当ARG作用于3D4/21细胞时，首先与细胞表面TLR6结合，激活TLR6/MyD88信号通路，而后活化NOX2型NADPH氧化酶，使细胞内ROS水平增加。ROS促进了细胞质中p38和ERK的磷酸化，进而提高了TNF-α与TGF-β1两种细胞因子的转录和分泌水平。同时，TLR6/MyD88信号通路的激活和ROS水平提高共同参与并调控3D4/21的吞噬活性的增强。结论，ARG能够通过TLR6/MyD88-NOX2 oxidase-MAPKs信号转导途径活化3D4/21巨噬细胞。本项目深入探索了ARG对3D4/21的免疫调节功能，并阐释了其作用机制。结果将为ARG提高畜禽机体免疫力的临床应用奠定理论基础，也将为推动中药在兽医临床方面的研究开发起到积极作用。