新疆特有植物巨车前保护急性肺损伤活性成分及作用机制研究

With high morbidity and high mortality characteristics, there is no specific treatment on Acute Lung Injury (ALI/ARDS). The treatment for this disease is still focus and difficulty in current clinical research. Plantago maxima is only distributed in Xinjiang in China. To the best of our knowledge, no phytochemical studies have been investigated on this plant. The project applicant found the extract of Plantago maxima has a certain role in the mitigation of LPS induced acute lung injury in mice in our preliminary study. On the basis of previous studies, LPS induced mouse model of acute lung injury and RAW264.7 cell model are used for screening anti ALI/ARDS activity fractions of Plantago maxima. Then the isolation and identification of active compounds are conducted by integrating a variety of chromatography and spectroscopy. And relevant biochemical indexes are also determined to disclose the mechanism of action. The launching of this project can not only enrich the natural product chemistry library, can also further exploit Xinjiang Plantago plant resources, and provide theoretical foundation and scientific basis for the development of innovative drugs of anti ACI/ARDS with our own intellectual property rights.

急性肺损伤(ALI/ARDS)具有高发病率和高病死率的特点，目前尚无特效治疗药物，针对该病的治疗仍是当今临床研究的重点和难点。巨车前在我国仅分布于新疆，关于巨车前的化学成分研究仍然是一个空白。项目申请人在前期研究中发现巨车前提取物对LPS诱导的小鼠急性肺损伤具有一定的缓解作用。本项目拟在前期研究的基础上，采用LPS诱导的小鼠急性肺损伤模型和RAW264.7细胞模型筛选巨车前抗ALI/ARDS活性部位，综合多种色谱技术和光谱技术，分离鉴定活性化合物，并测定相应生化指标，揭示巨车前抗ALI/ARDS活性的作用机制。本项目的开展不仅能够丰富天然产物化学库，也能进一步挖掘新疆车前属植物资源，为开发具有我国自主知识产权的抗急性肺损伤创新药物提供理论基础和科学依据。

ALI/ARDS的发病率及病死率较高，其治疗仍是当今临床研究的重点和难点，目前尚无特效治疗药物。此项目较系统地对巨车前活性部位中的化学成分进行分离鉴定与活性筛选研究，进而研究了活性化合物的抗ALI/ARDS作用机制，同时对主要活性成分的季节动态变化规律进行了探讨。运用多种色谱和波谱技术，从巨车前中分离鉴定26个化合物，包括1个含氯的新化合物和25个已知化合物，除木犀草素-7-O-β-D-吡喃葡萄糖苷和毛蕊花糖苷外，所有化合物均为首次从巨车前中分离得到。采用LPS诱导的Microglia BV-2细胞模型，对分离得到的单体化合物进行细胞水平活性筛选，明确了8个化合物为活性化合物。构建了Nrf-2转基因小鼠，与野生型小鼠在相同条件给予脂多糖，建立急性肺炎模型研究毛蕊花糖苷的作用机制，认为毛蕊花糖苷抗ALI/ARDS的作用可能是通过调控Nrf-2相关信号通路发挥作用的。采用LPS诱导的小鼠急性肺损伤模型探索木犀草素-7-O-β-D-吡喃葡萄糖苷作用机制，认为木犀草素-7-O-β-D-吡喃葡萄糖苷抗ALI/ARDS可能是通过调控TLR4相关信号通路发挥作用的。分析毛蕊花糖苷、异毛蕊花糖苷和木犀草素-7-O-β-D-吡喃葡萄糖苷这3种巨车前主要化学成分的动态变化规律，确定巨车前最佳采收期为6月。项目的开展丰富了天然产物化学库，为以后全面科学分析巨车前质量提供了依据，为阐明巨车前的药效物质基础和抗ALI/ARDS新药的研发提供了重要参考。