基于UPLC-Q/TOF-MS导向的菌类中药冬虫夏草化学成分及抗炎活性物质研究

Cordyceps sinensis is one of the Chinese Medicine treasures and also an important fungi resource. However, it is rare and precious. So, it is difficult to systematically study on chemical constituents of Cordyceps sinensis. The lack of further chemical and bioactive principles investigation of Cordyceps sinensis leads to the difficulty of quality control and authenticity identification. The success of the industrialization of Cordyceps sinensis, which is completely consistent with wild Cordyceps sinensis at chemical profiles and pharmacological activities, provide an chance for our research on chemical profiles and bioactive principles. The previous research of the applicant indicated that the breeding and wild Cordyceps sinensis showed equivalent inhibitory activities on IL-17 and TNF-α expression by GFP reporter cell lines method. Based on the above research, the chemical investigation of Cordyceps sinensis will be carried out with guidance of UPLC-Q/TOF-MS, the anti-inflammatory activities of chemical constituents of Cordyceps sinensis will be evaluated by GFP reporter cell lines method, and the specific chromatogram and authenticity criterion of Cordyceps sinensis will be establised. In this project, the chemical profile and anti-inflammatory principles of Cordyceps sinensis will be fully revealed for the first time, which will lay the foundation for scientific and comprehensive quality criterion. Meanwhile, it is possible to discover the anti-inflammatory lead compounds. The chemical investigation of rare and precious fungi can draw on the experience of UPLC-Q/TOF-MS method.

冬虫夏草是中华医药的瑰宝，也是重要的真菌资源，但其稀少珍贵，致使系统性地研究其化学成分困难，药效物质一直不清，导致现行质量标准无法全面反映其质量且真伪鉴别困难。冬虫夏草繁育品成功产业化且与野生冬虫夏草在化学指纹和药理活性上的高度一致性，使系统性地研究冬虫夏草化学成分和揭示其药效物质成为可能。申请人拟在前期利用炎症因子GFP报告细胞模型发现繁育品与野生冬虫夏草对IL-17和TNF-α均显示抑制活性的基础上，采用UPLC-Q/TOF-MS导向策略开展系统性的化学成分研究，运用炎症因子GFP报告细胞模型评价其抗炎活性，并建立冬虫夏草特征图谱和真伪鉴别标准。本项目将首次全面揭示菌类代表性中药冬虫夏草化学物质组全貌，阐明其抗炎的药效物质，为制定科学全面的质量标准奠定基础，并有望获得真菌来源抗炎药物先导化合物。UPLC-Q/TOF-MS导向策略也可为珍稀贵重真菌化学成分研究提供方法借鉴。

冬虫夏草是具有悠久历史文化，广泛公众认可度，独特资源分布和重要经济社会价值的名贵中药，被誉为中华医药瑰宝。由于资源稀缺贵重，至今仍缺乏系统深入的化学成分和药效物质研究，项目申报当年之前报道的有机小分子化合物仅有22个。本项目对15公斤冬虫夏草进行了系统的化学成分研究，为避免微量化学成分的分散丢失，我们采用了UPLC-Q/TOF-MS导向分离策略。本项目首次系统揭示了冬虫夏草中主含有机小分子化学成分全貌，完善了对冬虫夏草化学成分和结构类型的认识，构建了冬虫夏草主含有机小分子化学成分实物标本库，解决了冬虫夏草主含化学成分不明确的瓶颈，为后续研究如各种活性评价和质量标准建立奠定了基础。文献报道冬虫夏草中甾醇类化学成分具有抗炎活性，本项目首次从冬虫夏草中发现的脂肪酸甘油酯、脑苷脂、高度氧化不饱和脂肪酸均显示有抗炎活性，且是冬虫夏草主含化学成分，丰富了对冬虫夏草抗炎活性成分的认识。此外，本项目还首次发现冬虫夏草中脑苷脂类化学成分具有优异的免疫抑制活性，提示可能是其发挥免疫抑制功效的药效物质，为冬虫夏草质量标准完善和免疫抑制新药开发奠定了基础。