Universidad Andres Bello, Laboratorio de Biotecnología Molecular, Facultad de Ciencias de la Vida, 8370146, Santiago, Chile; Interdisciplinary Center for Aquaculture Research (INCAR), 4030000, Concepción, Chile.
Grupo de Marcadores Inmunológicos, Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile.
Vet Immunol Immunopathol. 2021 Jul;237:110240. doi: 10.1016/j.vetimm.2021.110240. Epub 2021 Apr 6.
Salmonids are a species of high commercial value in Chilean aquaculture, where muscle is the final product of the industry. Fish can be affected by stress during intensive cultures, increasing susceptibility to infections. Recently, we reported that muscle is an important focus of immune reactions. However, studies have shown the immunosuppressive effect of stress only in lymphoid organs, and few studies have been conducted on muscle and immunity. Hence, we determine the effects of cortisol on the immune-like response of fish myotubes challenged with Piscirickettsia salmonis by three trials. First, rainbow trout primary culture of muscle was cultured and treated with cortisol (100 ng/mL) for 3 and 4 h. Second, myotubes were challenged with P. salmonis (MOI 50) for 4, 6 and 8 h. And third, muscle cell cultures were pretreated with cortisol and then challenged with P. salmonis. The mRNA levels of glucocorticoid pathway and innate immunity were evaluated by qPCR. Cortisol increased the klf15 levels and downregulated the innate immune-related tlr5m gene and antimicrobial peptides. P. salmonis challenge upregulated several immune-related genes. Finally, cortisol pretreatment followed by P. salmonis challenge differentially modulated stress- and immune-related genes. These data suggest that fish muscle cells possess an intrinsic immune response and are differentially regulated by cortisol, which could lead to bacterial outbreaks in muscle under stress conditions.
鲑鱼是智利水产养殖中具有高商业价值的物种,肌肉是该行业的最终产品。鱼类在密集养殖过程中会受到压力的影响,增加感染的易感性。最近,我们报告称肌肉是免疫反应的重要焦点。然而,研究表明应激仅对淋巴器官具有免疫抑制作用,而对肌肉和免疫的研究很少。因此,我们通过三项试验确定皮质醇对鲑鱼鱼肌肉细胞受到鲑鱼鱼立克次氏体(Piscirickettsia salmonis)挑战时免疫样反应的影响。首先,培养虹鳟鱼原代肌肉培养物并用皮质醇(100ng/mL)处理 3 和 4 小时。其次,用 P. salmonis(MOI 50)挑战鱼肌肉细胞 4、6 和 8 小时。第三,用皮质醇预处理肌肉细胞培养物,然后用 P. salmonis 进行挑战。通过 qPCR 评估糖皮质激素途径和先天免疫的 mRNA 水平。皮质醇增加了 klf15 水平,下调了先天免疫相关的 tlr5m 基因和抗菌肽。P. salmonis 挑战上调了几个免疫相关基因。最后,皮质醇预处理后用 P. salmonis 进行挑战,差异调节了应激和免疫相关基因。这些数据表明,鱼类肌肉细胞具有内在的免疫反应,并且受到皮质醇的差异调节,这可能导致应激条件下肌肉中的细菌爆发。