Qiang Jun, Tao Fanyi, Bao Wenjin, He Jie, Liang Ming, Liang Cong, Zhu Haojun, Li Xiahong, Chen Deju, Xu Pao
Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, China.
Guangdong Wulonggang Aquatic Technology Development Co., Ltd., Guangzhou, China.
Front Physiol. 2019 Jul 5;10:868. doi: 10.3389/fphys.2019.00868. eCollection 2019.
Copper/zinc superoxide dismutase (Cu/Zn-SOD) plays critical roles in protecting cells and tissues against oxidative damage. Excessive copper ions (Cu) in water can damage the cells of aquatic organisms, leading to impaired growth and development and reduced antioxidant defenses. Many regulatory factors control the response to excess Cu. Among them, microRNAs (miRNAs) are important small RNAs that regulate the expression of their target genes and participate in the oxidative stress response. In the present study, we used bioinformatics and dual luciferase reporter gene analyses to demonstrate that the miR-489-3p of hybrid yellow catfish (♀ × ♂) binds to the 3'-untranslated region (UTR) of its target gene, which encodes a Cu/Zn-SOD. The regulatory relationship between this miRNA and its target gene was analyzed using qRT-PCR and luciferase activity assays. We also investigated the effect of the loss of miR-489-3p expression on the oxidative stress response of hybrid yellow catfish exposed to Cu. The 3'UTR region was found to be fully complementary to positions 2-9 of the 5'-end seed region of miR-489-3p. The miR-489-3p expression levels were negatively related to expression. Silencing of miR-489-3p up-regulated expression in the liver and gill tissues, increased activities of SOD and catalase, and reduced the malondialdehyde content. This study is the first to demonstrate that miR-489-3p targets to mediate the oxidative response to metal stress. These findings provide a theoretical basis for further studies on the response to oxidative stress caused by metals in cultured fish, and provide an experimental basis for the management of the culture environment.
铜锌超氧化物歧化酶(Cu/Zn-SOD)在保护细胞和组织免受氧化损伤方面发挥着关键作用。水中过量的铜离子(Cu)会损害水生生物的细胞,导致生长发育受损和抗氧化防御能力下降。许多调节因子控制着对过量铜的反应。其中,微小RNA(miRNA)是重要的小RNA,可调节其靶基因的表达并参与氧化应激反应。在本研究中,我们使用生物信息学和双荧光素酶报告基因分析来证明杂交黄颡鱼(♀×♂)的miR-489-3p与其靶基因的3'非翻译区(UTR)结合,该靶基因编码一种Cu/Zn-SOD。使用qRT-PCR和荧光素酶活性测定分析了这种miRNA与其靶基因之间的调控关系。我们还研究了miR-489-3p表达缺失对暴露于铜的杂交黄颡鱼氧化应激反应的影响。发现3'UTR区域与miR-489-3p 5'端种子区域的2-9位完全互补。miR-489-3p的表达水平与[此处原文缺失相关内容]的表达呈负相关。miR-489-3p的沉默上调了肝脏和鳃组织中的[此处原文缺失相关内容]表达,增加了超氧化物歧化酶和过氧化氢酶的活性,并降低了丙二醛含量。本研究首次证明miR-489-3p靶向[此处原文缺失相关内容]以介导对金属应激的氧化反应。这些发现为进一步研究养殖鱼类中金属引起的氧化应激反应提供了理论基础,并为养殖环境管理提供了实验依据。