文章摘要
金属蛋白酶组织抑制剂在皱纹盘鲍抗弧菌免疫中的作用研究
The role of tissue inhibitor of metalloproteinase in the anti-Vibrio immunity of Haliotis discus hannai
投稿时间:2020-10-04  修订日期:2021-01-27
DOI:
中文关键词: 皱纹盘鲍  副溶血弧菌  金属蛋白酶组织抑制剂  基质金属蛋白酶1  相互作用  先天性免疫
英文关键词: Haliotis discus hannai  Vibrio parahaemolyticus  Tissue inhibitor of metalloproteinase  Matrix metalloproteinase 1  Interaction  Innate immunity
基金项目:国家重点研发计划项目(2018YFD0901004);国家自然科学基金项目(31702372);国家贝类产业技术体系(CARS-49)
作者单位邮编
张明辉 集美大学 361021
章骞 集美大学 
林锦锦 集美大学 
张凌晶 集美大学 
刘光明 集美大学 
陈玉磊 集美大学 
曹敏杰 集美大学 
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中文摘要:
      【目的】本研究以皱纹盘鲍(Haliotis discus hannai)为研究对象,探讨金属蛋白酶组织抑制剂(Tissue inhibitor of Metalloproteinase, TIMP)在皱纹盘鲍抗弧菌免疫中的功能及其与基质金属蛋白酶1(Matrix metalloproteinase 1, MMP-1)相互作用关系。【方法】通过PCR技术克隆得到TIMP cDNA全长序列;利用荧光定量PCR技术分析了副溶血弧菌感染过程中皱纹盘鲍不同组织中TIMP的表达情况;利用RNA干扰技术分别敲低皱纹盘鲍TIMP和MMP-1的表达水平,研究TIMP与MMP-1在皱纹盘鲍抗弧菌免疫中的相互作用关系。【结论】克隆得到了TIMP的cDNA全长序列为2291 bp;在弧菌感染早期,TIMP在血淋巴细胞和鳃组织中的表达量均显著上调;鲍鱼体内MMP-1表达沉默后,TIMP表达显著上调;而TIMP表达受到抑制时,MMP-1表达量显著下降,说明TIMP表达受MMP-1负调控,而MMP-1表达受TIMP正调控。
英文摘要:
      As an important economical shellfish in coastal area of China, abalone is susceptible to bacterial infection, especially Vibiro parahemolyticus (V. parahemolyticus). Similar to other invertebrates, Haliotis discus hannai resist pathogen infection to the host by innate immune response. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous protein regulators of the matrix metalloproteinases (MMPs) family. As MMP-1 was engaged in the innate immunity of abalone, the involvement of TIMP in the regulation of MMP-1 expression and activity as well as the immune responses merits further investigation. In this study, the role of TIMP in the innate immune responses of H. discus hannai against V. parahaemolyticus infection and the interaction between TIMP and MMP-1 were investigated. The full length cDNA sequence of TIMP was obtained, and the expression of TIMP in different tissues of H. discus hannai were analyzed post Vibrio infection. The cloned TIMP cDNA sequence was 2291 bp in length. In the early stage of V. parahaemolyticus infection, the expression of TIMP in hemocytes and gill tissues were significantly up-regulated. To study the interaction between TIMP and MMP-1 in the anti-Vibrio immunity of abalone, RNA interference technology was used to knock down the expression levels of TIMP and MMP-1. After the silencing of MMP-1 gene in abalone, TIMP expression was significantly up-regulation. When TIMP expression was inhibited, MMP-1 expression decreased significantly. These results indicated that MMP-1 expression was positively regulated by TIMP, while TIMP expression was negatively regulated by MMP-1 in abalone. In conclusion, the present study established the interaction between TIMP and MMP-1 in abalone innate immunity against V. parahemolyticus infection. It also provided new insights for the prevention and control of pathogens in abalone culture.
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