MiRNAs-Modulation of Nrf2 Signaling Networks in Regulation Oxidative Stress of Chinese Perch Skeletal Muscle After Fasti
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ORIGINAL ARTICLE
MiRNAs-Modulation of Nrf2 Signaling Networks in Regulation Oxidative Stress of Chinese Perch Skeletal Muscle After Fasting Treatment Ping Wu 1,2 & Lin Chen 2 & Jia Cheng 2 & Yaxiong Pan 2 & Xinhong Guo 1 & Wuying Chu 2 & Jianshe Zhang 2 & Xuanming Liu 1 Received: 15 May 2020 / Accepted: 17 June 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Nrf2 is an important transcription factor involved in the antioxidant response and is widely expressed in animal tissues. The function of Nrf2 is regulated by its negative regulator Keap1 by inducing its cytoplasmic degradation. Recent studies have suggested that Nrf2 is also regulated post-transcriptionally via miRNAs. However, to date, how miRNAs regulate Nrf2 in fish skeletal muscles is unknown. In this study, the full-length cDNAs with 2398 bp of the Nrf2 was firstly cloned by SMART RACE amplification tools from Chinese perch. The Nrf2 gene structure and its 3’-UTR region for possible miRNA binding sites, as well as its spatial expression profile were assayed. Then, we employed TargetScan Fish tool MiRNAnome to predict putative sites for five miRNAs including miR-181a-5p, MiR-194a, MiR-216a, miR-459-5p, and miR-724. Using qRT-PCR assay, we found that Nrf2 mRNA levels have negative correlation with all five miRNAs expression in muscle of nutritionally deprived fish, and that ectopic expression of miR-181a-5p alone reduces Nrf2 mRNA levels. Luciferase reporter assay in a heterologous cell system revealed that each of the five miRNAs reduced Nrf2 expression, suggesting a direct regulatory mechanism. Moreover, the miR181a-5p suppression using specific antagomir led to a significant increase in Nrf2 expression in vivo. At the same time, the expression levels of the antioxidant enzymes CAT, ZnSOD, GPx, GSTA, and GSTA genes increased significantly after injecting miR-181a-5p antagomir. Taken together, these findings provide evidence that miRNAs are involved in the Nrf2 signaling networks in regulation of oxidative stress in fish, at least in Chinese perch muscle. Keywords Nrf2 . Chinese perch . Nutritional deprivation . Skeletal muscle . miRNAs . Keap1
Introduction Nrf2, an important transcription factor involved in the antioxidative response, is widely expressed in animal tissues, but is only activated through a series of oxidative and electrophilic stimuli (including ROS, some antioxidants, heavy metals, and certain disease processes) (Ma 2008, Kensler et al. 2007).
* Jianshe Zhang [email protected] * Xuanming Liu [email protected] 1
State Key laboratory of Chemo/Biosening and Chemomertrics, College of Biology, Hunan University, Changsha 410082, People’s Republic of China
2
Hunan Provincial Key Laboratory of Nutrition and Quality Control of Aquatic Animals, Department of Biological and Environmental Engineering, Changsha University, Changsha 410022, China
Excessive accumulation of ROS caused by oxidative stress increases cellular autophagy levels, including the antioxidant signaling pathway Nrf2-Keap1. In its static form,
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