缺氧,主要由全球变暖和水污染引发,已经成为一个备受关注的环境问题,对水生生态系统构成了极大的威胁。作为世界上最重要的经济鱼类之一,虹鳟鱼(Oncorhynchusmykiss)对低氧环境非常不耐受,然而,关于非编码RNA(ncRNAs)在虹鳟鱼对低氧胁迫的反应中的作用知之甚少。在这里,中度(Tm12L)和重度缺氧12h(Ts12L)和12h复氧对组织学的影响,生化参数(抗氧化剂,代谢和免疫)和转录组(lncRNA,研究了虹鳟鱼肝脏中的miRNA和mRNA)。我们通过双荧光素酶报告基因进一步验证了LOC110519952,new-m0023-5p和glut1a之间的调控关系,过表达和沉默测定。与Tm12L相比,Ts12L肝脏表现出更严重的氧化损伤。无氧,在低氧胁迫下,脂质和蛋白质代谢增强,特别是在Ts12L。我们还发现Tm12L可以增强先天免疫反应,在Ts12L中被抑制。此外,几个缺氧相关基因(glut1a,vegfaa,hmox,epea,foxo1a和igfbp1)和ceRNA网络从1824、427和545个差异表达的mRNA中鉴定出来,miRNAs和lncRNAs,包括LOC118965299-novel-m0179-3p-epea,LOC110519952-novel-m0023-5p-glut1a,MSTRG.7382.2-miR-184-y-hmox和LOC110520012-miR-206-y-vegfaa。通过体外和体内功能分析,我们证明了glut1a是new-m0023-5p的靶标,LOC110519952可以通过靶向new-m0023-5p来正向调节glut1a。LOC110519952的引入可以减弱new-m0023-5p对虹鳟鱼肝细胞活力和增殖的促进作用。本研究突出了不同浓度低氧胁迫下虹鳟鱼调控机制的差异,为进一步研究鱼类适应低氧环境的分子机制提供了有价值的数据。
Hypoxia, largely triggered by global warming and water contamination, has become an environmental issue of great concern, posing a great threat to aquatic ecosystem. As one of the world\'s most economically important fish, rainbow trout (Oncorhynchus mykiss) is extremely intolerant of hypoxic environments, however, little is known about the roles of non-coding RNAs (ncRNAs) in the response of rainbow trout to hypoxia stress. Herein, effects of moderate (Tm12L) and severe hypoxia for 12 h (Ts12L) and 12 h reoxygenation on histology, biochemical parameters (antioxidant, metabolism and immunity) and transcriptome (lncRNA, miRNA and mRNA) in rainbow trout liver were investigated. We further validated the regulatory relationships between LOC110519952, novel-m0023-5p and glut1a via dual‑luciferase reporter, overexpression and silencing assays. Compared with Tm12L, the liver in Ts12L showed more severe oxidative damage. Anaerobic, lipid and protein metabolism was enhanced under hypoxia stress, especially in Ts12L. We also found that Tm12L could strengthen innate immune response, which was inhibited in Ts12L. Besides, several hypoxia-related genes (glut1a, vegfaa, hmox, epoa, foxo1a and igfbp1) and ceRNA networks were identified from 1824, 427 and 545 differentially expressed mRNAs, miRNAs and lncRNAs, including LOC118965299-novel-m0179-3p-epoa, LOC110519952-novel-m0023-5p-glut1a, MSTRG.7382.2-miR-184-y-hmox and LOC110520012-miR-206-y-vegfaa. Through in vitro and in vivo functional analysis, we demonstrated that glut1a is a target of novel-m0023-5p, and LOC110519952 can positively regulate glut1a by targeting novel-m0023-5p. Introduction of LOC110519952 could attenuate the promoting effects of novel-m0023-5p on rainbow trout liver cell viability and proliferation. This study highlights the differences in the regulatory mechanism of rainbow trout under different concentrations of hypoxia stress and provides valuable data for further research on the molecular mechanisms of fish adaptation to hypoxic environments.