Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Science, Tianjin Normal University, Tianjin, 300387, People's Republic of China.
Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Science, Tianjin Normal University, Tianjin, 300387, People's Republic of China.
Fish Shellfish Immunol. 2020 Apr;99:301-309. doi: 10.1016/j.fsi.2020.02.021. Epub 2020 Feb 13.
Matrix metalloproteinases (MMPs) are a cluster of enzymes that degrade the extracellular matrix (ECM) and some intracellular proteins; as such, they play an important role in tissue regeneration, infant growth, animal reproduction, and immunity. Most research into MMPs focuses mainly on their effects on the mammalian immune system. However, it is not clear how MMPs affect immune processes in crustaceans. Here, we cloned the open reading frame (ORF) of Eriocheir sinensis (Chinese mitten crab) MMP-14 (EsMMP-14) to explore the role of MMPs in crustacean innate immune responses. RT-PCR results showed that stimulation of crab with LPS and poly I:C upregulated expression of EsMMP-14 markedly. Besides, following the stimulation of 20-Hydroxyecdysone, the expression level of EsMMP-14 increased robustly, suggesting that EsMMP-14 involved in the molt process of E. sinensis. Hematoxylin and eosin staining of hepatopancreas and intestine revealed that knocking down EsMMP-14 maintained morphology following infection by Bacillus thuringiensis. Moreover, downregulated expression of EsMMP-14 increased the survival rate of infected E. sinensis. These results show that EsMMP-14 plays a role in innate immune responses of E. sinensis and fills a gap in our knowledge about the function of MMPs in crustaceans.
基质金属蛋白酶（MMPs）是一类能够降解细胞外基质（ECM）和一些细胞内蛋白的酶，因此，它们在组织再生、婴儿生长、动物繁殖和免疫中发挥着重要作用。大多数关于 MMPs 的研究主要集中在它们对哺乳动物免疫系统的影响上。然而，目前还不清楚 MMPs 如何影响甲壳类动物的免疫过程。在这里，我们克隆了中华绒螯蟹（Chinese mitten crab）MMP-14（EsMMP-14）的开放阅读框（ORF），以探索 MMPs 在甲壳类动物先天免疫反应中的作用。RT-PCR 结果表明，LPS 和 poly I:C 刺激显著上调了 EsMMP-14 的表达。此外，在 20-羟基蜕皮酮的刺激下，EsMMP-14 的表达水平显著增加，这表明 EsMMP-14 参与了中华绒螯蟹的蜕皮过程。肝胰腺和肠道的苏木精和伊红染色表明，敲低 EsMMP-14 可以维持感染苏云金芽孢杆菌后的形态。此外，下调 EsMMP-14 的表达可提高感染中华绒螯蟹的存活率。这些结果表明，EsMMP-14 在中华绒螯蟹的先天免疫反应中发挥作用，填补了我们对 MMPs 在甲壳类动物中的功能的认识空白。
