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Abstract：
BACKGROUND: Cystic echinococcosis (CE) is a widespread zoonosis caused by the infection with Echinococcus granulosus sensu lato (E. granulosus s.l.). CE cysts mainly develop in the liver of intermediate hosts, characterized by the fibrotic tissue that separates host organ from parasite. However, precise mechanism underlying the formation of fibrotic tissue in CE remains unclear.
METHODS: To investigate the potential impact of ubiquitin-conjugating enzymes on liver fibrosis formation in CE, two members of ubiquitin-conjugating (UBC) enzyme of Echinococcus granulosus (EgE2D2 and EgE2N) were recombinantly expressed in Escherichia coli and analyzed for bioinformatics, immunogenicity, localization, and enzyme activity. In addition, the secretory pathway and their effects on the formation of liver fibrosis were also explored.
RESULTS: Both rEgE2D2 and rEgE2N possess intact UBC domains and active sites, exhibiting classical ubiquitin binding activity and strong immunoreactivity. Additionally, EgE2D2 and EgE2N were widely distributed in protoscoleces and germinal layer, with differences observed in their distribution in 25-day strobilated worms. Further, these two enzymes were secreted to the hydatid fluid and CE-infected sheep liver tissues via a non-classical secretory pathway. Notably, TGFβ1-induced LX-2 cells exposed to rEgE2D2 and rEgE2N resulted in increasing expression of fibrosis-related genes, enhancing cell proliferation, and facilitating cell migration.
CONCLUSIONS: Our findings suggest that EgE2D2 and EgE2N could secrete into the liver and may interact with hepatic stellate cells, thereby promoting the formation of liver fibrosis.
METHODS: To investigate the potential impact of ubiquitin-conjugating enzymes on liver fibrosis formation in CE, two members of ubiquitin-conjugating (UBC) enzyme of Echinococcus granulosus (EgE2D2 and EgE2N) were recombinantly expressed in Escherichia coli and analyzed for bioinformatics, immunogenicity, localization, and enzyme activity. In addition, the secretory pathway and their effects on the formation of liver fibrosis were also explored.
RESULTS: Both rEgE2D2 and rEgE2N possess intact UBC domains and active sites, exhibiting classical ubiquitin binding activity and strong immunoreactivity. Additionally, EgE2D2 and EgE2N were widely distributed in protoscoleces and germinal layer, with differences observed in their distribution in 25-day strobilated worms. Further, these two enzymes were secreted to the hydatid fluid and CE-infected sheep liver tissues via a non-classical secretory pathway. Notably, TGFβ1-induced LX-2 cells exposed to rEgE2D2 and rEgE2N resulted in increasing expression of fibrosis-related genes, enhancing cell proliferation, and facilitating cell migration.
CONCLUSIONS: Our findings suggest that EgE2D2 and EgE2N could secrete into the liver and may interact with hepatic stellate cells, thereby promoting the formation of liver fibrosis.
摘要：
背景：囊性包虫病（CE）是一种由细粒棘球蚴感染引起的广泛的人畜共患病（E.granulosuss.l.).CE囊肿主要在中间宿主的肝脏中发展，以将宿主器官与寄生虫分开的纤维化组织为特征。然而,CE纤维化组织形成的确切机制尚不清楚。
方法：为了研究泛素结合酶对CE肝纤维化形成的潜在影响，在大肠杆菌中重组表达了细粒棘球蚴泛素结合酶(UBC)的两个成员(EgE2D2和EgE2N),并进行了生物信息学分析,免疫原性,本地化,和酶活性。此外,还探讨了其分泌途径及其在肝纤维化形成中的作用。
结果：rEgE2D2和rEgE2N都拥有完整的UBC域和活性位点，表现出经典的泛素结合活性和强的免疫反应性。此外,EgE2D2和EgE2N广泛分布在原头骨和生发层中，在25天的频闪蠕虫中观察到它们的分布差异。Further,这两种酶通过非经典分泌途径分泌到包虫液和CE感染的绵羊肝组织中。值得注意的是,TGFβ1诱导的LX-2细胞暴露于rEgE2D2和rEgE2N导致纤维化相关基因表达增加,增强细胞增殖，促进细胞迁移。
结论：我们的研究结果表明，EgE2D2和EgE2N可以分泌到肝脏，并可能与肝星状细胞相互作用，从而促进肝纤维化的形成。
方法：为了研究泛素结合酶对CE肝纤维化形成的潜在影响，在大肠杆菌中重组表达了细粒棘球蚴泛素结合酶(UBC)的两个成员(EgE2D2和EgE2N),并进行了生物信息学分析,免疫原性,本地化,和酶活性。此外,还探讨了其分泌途径及其在肝纤维化形成中的作用。
结果：rEgE2D2和rEgE2N都拥有完整的UBC域和活性位点，表现出经典的泛素结合活性和强的免疫反应性。此外,EgE2D2和EgE2N广泛分布在原头骨和生发层中，在25天的频闪蠕虫中观察到它们的分布差异。Further,这两种酶通过非经典分泌途径分泌到包虫液和CE感染的绵羊肝组织中。值得注意的是,TGFβ1诱导的LX-2细胞暴露于rEgE2D2和rEgE2N导致纤维化相关基因表达增加,增强细胞增殖，促进细胞迁移。
结论：我们的研究结果表明，EgE2D2和EgE2N可以分泌到肝脏，并可能与肝星状细胞相互作用，从而促进肝纤维化的形成。
