Zhou Keli, Gao Lei, Ge Pingan, Wang Ling, Liu Lu, Ye Jiayu, Xu Hairu, Wang Lingling, Song Linsheng
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China.
Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Zhuhai, 519000, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China; Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China.
Dev Comp Immunol. 2025 Jan;162:105306. doi: 10.1016/j.dci.2024.105306. Epub 2024 Dec 20.
miRNA, a type of endogenous small non-coding RNA, is involved in the response to various environmental stresses through post-transcriptional regulation. In the present study, the role of CgmiR307 in the regulation of oxidative response under high-temperature stress by targeting CgNrf2 was investigated in the Pacific oyster Crassostrea gigas. The binding site of CgmiR307 were predicted at 1799-1818 bp in the 3'-UTR of CgNrf2, and the binding activity of CgmiR307 with the mRNA of CgNrf2 was further proved by the dual-luciferase reporter assay. The expression levels of CgmiR307 and CgNrf2 in gill were significantly higher than in other tissues, and exhibited significant fluctuations and variations after exposure to 28 °C. There was a significant reduction in the expression levels of CgSOD, and CgCAT in gill, as well as the activities of SOD, CAT, and T-AOC, while ROS and MDA contents significantly increased in CgNrf2-RNAi oysters. After CgmiR307 agomir injection and high-temperature stress, the expression levels of CgNrf2, CgSOD and CgCAT in gill, the activities of SOD and CAT and T-AOC decreased significantly, while ROS and MDA content significantly increased. After CgmiR307 antagomir injection and high-temperature stress, the changes in the parameters of oxidative response shown exactly the opposite trend. These results demonstrated that CgmiR307 was involved in the regulation of oxidative response by inhibiting the mRNA expression of CgNrf2 under high-temperature stress.
微小RNA(miRNA)是一种内源性小非编码RNA,通过转录后调控参与对各种环境胁迫的响应。在本研究中,在太平洋牡蛎(Crassostrea gigas)中研究了CgmiR307通过靶向CgNrf2在高温胁迫下对氧化应激反应的调控作用。预测CgmiR307在CgNrf2的3'-UTR中1799 - 1818 bp处有结合位点,双荧光素酶报告基因检测进一步证实了CgmiR307与CgNrf2 mRNA的结合活性。鳃中CgmiR307和CgNrf2的表达水平显著高于其他组织,在暴露于28°C后呈现出显著的波动和变化。在CgNrf2 - RNA干扰的牡蛎中,鳃中CgSOD和CgCAT的表达水平显著降低,同时SOD、CAT和总抗氧化能力(T - AOC)的活性降低,而活性氧(ROS)和丙二醛(MDA)含量显著增加。注射CgmiR307激动剂并进行高温胁迫后,鳃中CgNrf2、CgSOD和CgCAT的表达水平,SOD、CAT的活性以及T - AOC显著降低,而ROS和MDA含量显著增加。注射CgmiR307拮抗剂并进行高温胁迫后,氧化应激反应参数的变化呈现出完全相反的趋势。这些结果表明,在高温胁迫下,CgmiR307通过抑制CgNrf2的mRNA表达参与氧化应激反应的调控。