药用蜈蚣蛋白质肽类药效分子群的鉴别及其药理学作用机制

Centipedes are one of the most important traditional Chinese medicinal materials coming from animals. However, owing to lacking a comprehensive recognition of bioactive fundamentals existed in clinic used centipedes, especially bioactive components contained in their venoms, the clinic used centipedes varied both in species (the most common used centipedes including Scolopendra subspinipes multilans、S. subspinipes multidens、S. mojiangica Zhang et Chi and S. negrocapitis) and in used manner (whole body and decapitate animal). We have already characterized approximately 400 novel protein/peptide molecules from the venom of S. subspinipes dehaani and systematically investigated relationships between their molecular structures and pharmacological properties. Our results revealed that the molecular structures of ion-channel acting components from S. subspinipes dehaani are totally different from that from S. subspinipes multilans, no identical ion-channel acting molecule was found in these 2 centipede species. Our results, in molecular level, confirmed that different bioactive fundamental groups exist in different centipede species. Thus, standardization the clinic usage of centipedes is both inevitable and necessary. Based on our previous works, this research project will comprehensively reveal the molecular properties of proteins/peptides in 4 common used centipede species. Meanwhile, we will extensively and deeply carry out systematical investigation to identify bioactive fundamental groups of whole body, venom and torso in 2 representative centipede species, namely S. subspinipes multilans and S. subspinipes dehaani by a combination of pharmacological activity assays and bioinformatic analyses. Our results will provide useful information to interpret the material base and functional characterization of centipede bioactive groups and provide scientific basis for safe, efficiency and reasonable use of clinic used centipedes. Carried out the present project will lay a solid foundation for the research of innovative medicines based on novel centipede bioactive molecules, encourage TCM modernization. The proposed project has both scientific significance and of great value in clinic.

蜈蚣是一味传统的重要动物源中药材。对药用蜈蚣药效物质，特别是毒液组分缺乏全面的认识是目前蜈蚣药用从种类(少棘蜈蚣、多棘蜈蚣、墨江蜈蚣、黑头蜈蚣)和方式(整体、去头)存在差异的重要原因。我们已从哈氏蜈蚣毒液中新识别400余种蛋白/肽类活性物质，系统地研究了分子结构与药理学活性的关系；研究揭示来源于哈氏蜈蚣和少棘蜈蚣离子通道毒素的分子结构完全不同，从分子水平上证明了药用蜈蚣之间存在不同的药效分子群。因此，规范化蜈蚣应用势在必行。立足前期工作，本项目将全面揭示4种药用蜈蚣的蛋白质/肽类分子特征，进一步通过药理学活性和生物信息学手段，深入开展2个代表物种－少棘蜈蚣和墨江蜈蚣全体、毒液和躯干药效分子的辨识，诠释其药效物质基础及其作用特点，为不同类型蜈蚣安全、有效、合理使用，提供科学依据。同时，项目的实施将为基于蜈蚣药效分子的创新药物研发奠定坚实基础，促进中药代现代化发展，具有重要的科学意义和应用价值

蜈蚣是最古老的有毒动物之一，用其毒液作为捕食或防御武器；它们的毒液成分复杂，具有不同的生物医学活性和药理学活性；但是，几乎没有研究提到蜈蚣毒液中毒素类似蛋白或多肽的广谱性。在本研究中，我们采用蛋白质组-转录组学相结合的方法揭示了少棘蜈蚣（Scolopendra subspinipes mutilans）中毒素类似蛋白的多样性，通过ESI-MS/MS质谱鉴定和转录组预测分别从少棘蜈蚣毒液和躯干组织中鉴定到923种和6736种多肽；最终我们从其蛋白质组中鉴定到1369种特异蛋白，包括毒液和躯干共表达的100种蛋白；其中，至少有40种经鉴定是毒液毒素类似蛋白。同时，通过转录组数据挖掘鉴定出超过10倍的毒素类似蛋白，具备成熟毒素类似多肽的前体结构特征。最重要的是，整合转录组学和蛋白质组学分析发现，25种毒素类似蛋白/多肽（50%为神经毒素）在毒腺以外的其他部位仍有表达，并且参与了基因招募过程。这些结果展现了蜈蚣毒素类似蛋白的多样性，并为蜈蚣毒素类似蛋白用于医学制药提供了新的依据。.药典品种少棘蜈蚣与临床替代品墨江蜈蚣毒液的比较分析结果揭示二者的毒液在蛋白质和多肽层面有较高的相似性，但仍存在巨大差异。在多肽组层面，少棘蜈蚣与墨江蜈蚣毒液分别存在38个特有的多肽分子，少棘蜈蚣的16条多肽与墨江蜈蚣的14条多肽组成了一个由30个分子形成的类似肽；在蛋白质层面，少棘蜈蚣与墨江蜈蚣毒液分别存在120和293个特有的蛋白质分子，少棘蜈蚣的192个蛋白质与墨江蜈蚣的218个蛋白质组成了一个由410个分子形成的类似蛋白。揭示了不同蜈蚣物种在临床的治疗中需进行进一步的深入评估。