IOCAS-IR
沙海蜇毒素金属蛋白酶致炎作用研究
李翱宇
Subtype硕士
Thesis Advisor于华华
2019-05
Degree Grantor中国科学院大学
Place of Conferral中国科学院海洋研究所
Degree Name工程硕士
Keyword沙海蜇毒素 金属蛋白酶 水母皮炎 肌肉毒性
Abstract

近年来,随着海洋环境变化和人类生产生活等的影响,导致水母在世界范围内暴发的现象频繁发生,水母蜇伤已成为人们重点关注的公共安全问题之一。水母皮炎是水母蜇伤后的主要临床反应,炎症反应伴随了水母皮炎的整个过程,严重影响了人们的日常生活和健康。由于水母毒素成分的复杂多样,使得至今未能有治疗水母皮炎的有效药物。通过对水母毒素的组学研究,表明水母毒素中含有丰富的金属蛋白酶成分,研究表明,金属蛋白酶成分在陆生动物的毒液中发挥了重要的作用。因此,本文以中国常见的大型蜇人水母沙海蜇为研究对象,分别从体外和体内层面,通过基质金属蛋白酶抑制剂对毒素诱导的炎症反应的抑制作用,探究了金属蛋白酶对水母皮炎的生物学效应,主要结果如下:

1. 体外水平的研究表明,沙海蜇毒素能够显著降低细胞的存活率,并且能通过促进细胞凋亡导致细胞死亡;毒素作用于两种皮肤细胞和两种炎症模型细胞时,细胞中的三种炎症因子IL-6MCP-1TNF-α的基因表达水平和蛋白表达水平都显著增高,说明毒素能够刺激细胞产生炎症反应。当金属蛋白酶抑制剂BMTEDTA与沙海蜇毒素反应后,能够显著降低毒素对细胞的毒性作用,提高细胞的存活率,细胞中的炎症因子的含量也显著下降。说明沙海蜇毒素中的金属蛋白酶在炎症的发生过程中起重要作用。

2. 体内研究表明,沙海蜇毒素能够刺激小鼠体内两种炎症细胞-淋巴细胞和白细胞的产生,以及三种炎症因子IL-6MCP-1TNF-α的表达显著增加。皮肤组织病理切片表明沙海蜇毒素导致小鼠皮肤组织中炎症细胞的含量明显增多;而金属蛋白酶抑制剂与毒素作用后,皮肤组织中炎症细胞的含量减少,表明沙海蜇毒素中金属蛋白酶具有致炎作用。

3. 沙海蜇毒素的肌肉毒性实验表明,沙海蜇毒素能够导致机体中乳酸脱氢酶和肌酸激酶活性的显著升高,可能会造成肌肉组织损伤,而金属蛋白酶抑制剂作用后的毒素对机体中乳酸脱氢酶和肌酸激酶的活性显著降低,说明金属蛋白酶在沙海蜇毒素的肌肉毒性中发挥了重要作用。

Other Abstract

In recent years, with the changes of marine environment and the impact of human production and life, the phenomenon of jellyfish bloom around the world has occurred frequently. Jellyfish dermatitis is the main clinical response after jellyfish stings. The inflammatory response accompanied by the entire process of jellyfish dermatitis, seriously affecting people's daily life and health. Due to the complexity and variety of jellyfish venom, there has been no effective drug for the treatment of jellyfish dermatitis. The histological study of jellyfish venom shows that there are abundant metalloproteinases in jellyfish venom. The study shows that metalloproteinases play an important role in the venom of terrestrial animals. Therefore, in this paper, the biological effects of metalloproteinases on jellyfish dermatitis were studied in vitro and in vivo by using matrix metalloproteinase inhibitors to inhibit the inflammation, respectively. The main results are as follows:

1. In vitro studies have shown that NnNV can significantly reduce cell survival and induce cell death by promoting cell apoptosis. When the NnNV acts on two kinds of skin cells and two kinds of inflammatory model cells, the gene expression and protein expression levels of IL-6, MCP-1 and TNF-α in cells are significantly increased, indicating that NnNV can stimulate cells produced inflammation. When metalloproteinase inhibitors, BMT and EDTA, react with NnNV, they can significantly reduce the toxicity of NnNV to cells, improve the survival rate of cells, and significantly reduce the content of inflammatory factors in cells. It is indicated that the metalloproteinase in the NnNV plays an important role in the process of inflammation.

2. In vivo studies showed that NnNV could stimulate the expression of two inflammatory cells, lymphocyte and leukocyte, and three inflammatory factors, IL-6, MCP-1 and TNF-α in mice. Skin histopathological sections showed that NnNV had an effect on inflammatory cells, and the content of inflammatory cells in mice skin increased significantly when metalloproteinase inhibitor acted. Afterwards, the content of inflammatory cells in skin tissue decreased, indicating that metalloproteinases in NnNV have an inflammatory effect.

3. Muscle toxicity test of NnNV showed that NnNV could increase the activity of lactate dehydrogenase and creatine kinase in vivo, which might cause muscle tissue injury. However, the activity of lactate dehydrogenase and creatine kinase decreased significantly after the action of metalloproteinase inhibitor, which indicated that metalloproteinase played an important role in the myotoxicity of NnNV.

MOST Discipline Catalogue工学 ; 工学::生物工程
Language中文
Document Type学位论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/156892
Collection中国科学院海洋研究所
实验海洋生物学重点实验室
Recommended Citation
GB/T 7714
李翱宇. 沙海蜇毒素金属蛋白酶致炎作用研究[D]. 中国科学院海洋研究所. 中国科学院大学,2019.
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