沙海蜇毒素金属蛋白酶对水母皮炎的生物学效应及致炎机理研究

In recent years, the frequent occurrence of jellyfish sting brings serious public health problems and jellyfish dermatitis has become common Endemic Diseases in coastal areas. Jellyfish venom is the key cause of dermatitis. However, it is unclear that the exact component in jellyfish venom is responsible for dermatitis. Metalloproteinase was rich in jellyfish venom and it might be responsible for dermatitis according to our pre-studies on the enzymatic characteristics and the effect of specific inhibitor of metalloproteinase on the cutaneous vascular permeability. However, the biological effect on jellyfish dermatitis and the inflammatory mechanism of metalloproteinase from jellyfish venom are still unclear. Therefore, in this study, we will purify and characterize metalloproteinase from venom of jellyfish Nemopilema nomurai; and elucidate its biological effect on dermatitis by analyzing the histopathological slides, vascular permeability and releasing levels of inflammatory factors; and further reveal the inflammatory mechanism of metalloproteinase. This study will provide the theoretical basis for the treatment of jellyfish dermatitis and severe sting, as well as the strategy for the large-scale outbreak of jellyfish in view of venom control. In addition, a method reference for a convenient and accurate model to evaluate jellyfish dermatitis in vitro will be established to improve the level of screening antidote against jellyfish venom and help for drug research & development of jellyfish dermatitis.

近年来，水母蜇伤频繁发生，水母皮炎已成为沿海地区常见的“地方病”。水母毒素是引发水母皮炎的物质基础，但毒素中哪种成分与皮炎发生相关并不清楚。水母毒素中含有丰富的金属蛋白酶，基于毒素酶学性质及金属蛋白酶特异性抑制剂对皮肤血管通透性的影响，我们推测金属蛋白酶在水母蜇伤引发的皮肤炎症中可能起到关键性作用。但金属蛋白酶对水母皮炎的生物学效应和致炎机理均不清楚。本项目拟以“海洋杀手”沙海蜇为研究对象，对沙海蜇毒素金属蛋白酶进行纯化与表征；从组织病理、炎症因子、血管通透性等方面明确金属蛋白酶对水母皮炎的生物学效应；基于代谢组学、蛋白组学及炎症通路分析，阐明金属蛋白酶的致炎机理。通过本项目的研究，为水母皮炎及重症蜇伤患者的治疗提供理论依据；为应对水母大规模暴发提供毒素防治策略；为建立一套方便、准确的水母皮炎体外评价体系提供方法学依据，提升水母毒素解毒剂的筛选水平，助力水母皮炎药物的研制。

近年来，水母蜇伤频繁发生，水母皮炎已成为沿海地区常见的“地方病”，严重影响着滨海旅游、渔业捕捞和军事训练等涉海活性。水母毒素是引发水母皮炎的物质基础，但毒素中哪种成分与皮炎发生相关并不清楚。水母毒素中含有丰富的金属蛋白酶，基于毒素酶学性质及金属蛋白酶特异性抑制剂对皮肤血管通透性的影响，我们推测金属蛋白酶在水母蜇伤引发的皮肤炎症中可能起到关键性作用。但金属蛋白酶对水母皮炎的生物学效应和致炎机理均不清楚。本项目以“海洋杀手”沙海蜇为研究对象，利用硫酸铵沉淀、离子交换层析和凝胶色谱层析，对沙海蜇毒素的金属蛋白酶成分进行了纯化，得到4个具有金属蛋白酶活性的组分，发现蛋白酶活性的协同调控作用；通过细胞和动物实验明确了沙海蜇毒素金属蛋白酶能够导致细胞炎症因子IL-6，MCP-1和TNF-α的基因和蛋白表达上调，使小鼠的皮肤通透性增大，导致皮炎的发生；利用皮肤蛋白质组-血清代谢组，系统解析了水母毒素致炎机制，沙海蜇毒素通过降解细胞黏附分子破坏组织细胞的结构和功能、影响通道蛋白结合ATP，破坏渗透平衡造成局部组织急性炎症，并引发PI3K/Akt激活促进免疫细胞活化和细胞因子释放、Intergrin alpha-M上调促进的免疫细胞游出等促炎效应。本项目发表SCI论文9篇，申请国家发明专利3项，其中1项已获得授权，培养研究生2名，项目负责人获第十二届青岛市青年科技奖。通过本项目的研究，确定了沙海蜇毒素金属蛋白酶对水母皮炎的生物学效应及致炎机理,为抗水母毒素化合物的筛选、水母蜇伤的防治及水母暴发的防灾减灾提供理论依据。