State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, School of Marine Sciences, Ningbo University, Ningbo 315211, China; Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315211, China; Key Laboratory of Applied Marine Biotechnology of Ministry of Education, Ningbo University, Ningbo 315211, China.
State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, School of Marine Sciences, Ningbo University, Ningbo 315211, China; Laboratory of Biochemistry and Molecular Biology, School of Marine Sciences, Ningbo University, Ningbo 315211, China; Key Laboratory of Applied Marine Biotechnology of Ministry of Education, Ningbo University, Ningbo 315211, China.
Int J Biol Macromol. 2024 Oct;278(Pt 1):134624. doi: 10.1016/j.ijbiomac.2024.134624. Epub 2024 Aug 10.
The molecular chaperone GroEL, commonly found in various bacterial species, exhibits heightened expression levels in response to high temperatures and increased levels of oxygen free radicals. Limited literature currently exists on the probiotic role of GroEL in invertebrates. This study sought to explore how the surface protein GroEL from Lactobacillus plantarum Ep-M17 impacts the intestinal barrier function of Penaeus vannamei. Through pull-down and immunofluorescence assays, the interaction between GroEL and Act1 in the gastrointestinal tract of P. vannamei was confirmed. Results from bacterial binding assays demonstrated that rGroEL can bind to pathogens like Vibrio parahaemolyticus E1 (V. p-E1). In vitro experiments revealed that the administration of rGroEL significantly decreased the levels of inflammatory cytokines induced by pathogens while preserving the integrity of tight junctions between intestinal epithelial cells and reducing bacteria-induced apoptosis. Additionally, rGroEL notably lessened the intestinal loading of V. p-E1 in P. vannamei, downregulated immune-related gene expression, and upregulated BCL/BAX expression in the intestines following V. p-E1 challenge. Mechanistic investigations further showed that rGroEL treatment effectively suppressed the expression and phosphorylation of proteins involved in the NF-κB and PI3K-AKT-mTOR signalling pathways in the intestines of bacteria-infected P. vannamei. Furthermore, GroEL reinforces protection against bacterial infections by enhancing the phagocytic and anti-apoptotic capabilities of P. vannamei hemocytes. These results suggest that GroEL may impede the interaction between pathogens and the intestinal mucosa through its competitive binding characteristics, ultimately reducing bacterial infections.
热休克蛋白 GroEL 普遍存在于各种细菌物种中,其表达水平会随着温度升高和氧自由基水平增加而升高。目前关于 GroEL 在无脊椎动物中的益生菌作用的文献有限。本研究旨在探索植物乳杆菌 Ep-M17 的表面蛋白 GroEL 如何影响凡纳滨对虾的肠道屏障功能。通过下拉和免疫荧光分析,证实了 GroEL 与凡纳滨对虾胃肠道中 Act1 的相互作用。细菌结合实验结果表明,rGroEL 可以与副溶血弧菌 E1(V. p-E1)等病原体结合。体外实验表明,rGroEL 的给药可显著降低病原体诱导的炎症细胞因子水平,同时保持肠上皮细胞之间紧密连接的完整性,并减少细菌诱导的细胞凋亡。此外,rGroEL 显著减少了凡纳滨对虾中 V. p-E1 的肠道负荷,下调了 V. p-E1 感染后对虾肠道中免疫相关基因的表达,并上调了 BCL/BAX 的表达。机制研究进一步表明,rGroEL 处理可有效抑制 NF-κB 和 PI3K-AKT-mTOR 信号通路相关蛋白在细菌感染对虾肠道中的表达和磷酸化。此外,GroEL 通过增强对虾血细胞的吞噬和抗凋亡能力来增强对细菌感染的保护作用。这些结果表明,GroEL 可能通过其竞争性结合特性阻碍病原体与肠道黏膜的相互作用,从而减少细菌感染。