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缺氧暴露的抗氧化能力、与能量代谢相关的酶活性及转录组分析

Antioxidant Capacity, Enzyme Activities Related to Energy Metabolism, and Transcriptome Analysis of Exposed to Hypoxia.

作者信息

He Pingping, Li Wei, Wei Pinyuan, Jiang Linyuan, Guan Junliang, Ma Yuan, Zhang Li, Chen Yongxian, Zheng Yusi, Zhang Xingzhi, Peng Jinxia

机构信息

Guangxi Key Laboratory of Aquatic Genetic Breedingand Healthy Aquaculture, China (Guangxi)-ASEAN Key Laboratory of Comprehensive Exploitation and Utilization of Aquatic Germplasm Resources, Ministry of Agriculture and Rural Affairs, Guangxi Academy of Fishery Sciences, Nanning 530021, China.

出版信息

Antioxidants (Basel). 2024 Aug 30;13(9):1063. doi: 10.3390/antiox13091063.

Abstract

() is one of the three most commonly cultivated oyster species in China. Seasonal hypoxia is one of the most serious threats to its metabolism, reproductive behavior, and survival. To investigate the effects of hypoxia stress on the antioxidant capacity and energy metabolism of , the total antioxidant capacity (T-AOC), glycogen content, and enzyme activities (phosphofructokinase, PFK; pyruvate kinase, PK; phosphoenolpyruvate carboxykinase, PEPCK) of oysters were determined under normoxic (DO 6 ± 0.2 mg/L) and hypoxic (DO 1.5 mg/L) conditions at 0 h, 6 h, 48 h, and 72 h. We also determined the T-AOC, glycogen content, and enzyme activities of oysters under reoxygenation (recovered to normoxia for 24 h). To further examine the potential molecular regulatory mechanism of hypoxic adaptation, a transcriptome analysis was conducted on the gill of under normoxia (N, 72 h), hypoxia (H, 72 h), and reoxygenation (R). After being exposed to hypoxia for 6 h, the T-AOC, glycogen content, and enzyme activities of PK, PFK, and PEPCK in were significantly decreased. However, after prolonging the duration of hypoxia exposure for 72 h, the T-AOC, glycogen content, and enzyme activities increased compared to that of 48 h. After 24 h reoxygenation, the T-AOC, glycogen content, and enzyme activity of PK and PFK returned to close to initial levels. In addition, a transcriptome analysis discovered 6097 novel genes by mapping the genome with the clean reads. In total, 352 differentially expressed genes (DEGs) were identified in the H vs. N comparison group (235 upregulated and 117 downregulated genes). After recovery to normoxia, 292 DEGs (134 upregulated and 158 downregulated genes) were identified in the R vs. N comparison group, and 632 DEGs were identified (253 upregulated and 379 downregulated genes) in the R vs. H comparison group. The DEGs included some hypoxia-tolerant genes, such as phosphoenolpyruvate carboxykinase (), mitochondrial (), tyramine beta-hydroxylase (), superoxide dismutase (), glutathione S-transferase (), and egl nine homolog 1 isoform X2 (). Additionally, DEGs were significantly enriched in the KEGG pathways that are involved in hypoxia tolerance, including the metabolism of xenobiotics by cytochrome P450 pathways and the HIF-1 signaling pathway. Then, we selected the five hypoxic-tolerant candidate DEGs for real-time quantitative polymerase chain reaction (RT-qPCR) validation, and the results were consistent with the transcriptome sequencing data. These discoveries have increased our understanding of hypoxia tolerance, recovery ability after reoxygenation, and molecular mechanisms governing the responses to hypoxia in .

摘要

()是中国最常养殖的三种牡蛎品种之一。季节性缺氧是对其新陈代谢、生殖行为和生存最严重的威胁之一。为了研究缺氧胁迫对()抗氧化能力和能量代谢的影响,在常氧(溶解氧6±0.2毫克/升)和缺氧(溶解氧1.5毫克/升)条件下,于0小时、6小时、48小时和72小时测定了牡蛎的总抗氧化能力(T-AOC)、糖原含量和酶活性(磷酸果糖激酶、PFK;丙酮酸激酶、PK;磷酸烯醇式丙酮酸羧激酶、PEPCK)。我们还测定了复氧(恢复到常氧24小时)后牡蛎的T-AOC、糖原含量和酶活性。为了进一步研究缺氧适应的潜在分子调控机制,对常氧(N,72小时)、缺氧(H,72小时)和复氧(R)条件下的()鳃进行了转录组分析。暴露于缺氧6小时后,()的T-AOC、糖原含量以及PK、PFK和PEPCK的酶活性显著降低。然而,延长缺氧暴露时间至72小时后,与48小时相比,T-AOC、糖原含量和酶活性增加。复氧24小时后,PK和PFK的T-AOC、糖原含量和酶活性恢复到接近初始水平。此外,转录组分析通过将干净读段映射到()基因组发现了6097个新基因。总共,在H与N比较组中鉴定出352个差异表达基因(DEG)(235个上调基因和117个下调基因)。恢复到常氧后,在R与N比较组中鉴定出292个DEG(134个上调基因和158个下调基因),在R与H比较组中鉴定出632个DEG(253个上调基因和379个下调基因)。这些DEG包括一些耐缺氧基因,如磷酸烯醇式丙酮酸羧激酶()、线粒体()、酪胺β-羟化酶()、超氧化物歧化酶()、谷胱甘肽S-转移酶()和egl九同源物1异构体X2()。此外,DEG在参与耐缺氧的KEGG途径中显著富集,包括细胞色素P450途径对外源生物的代谢和HIF-1信号通路。然后,我们选择了五个耐缺氧候选DEG进行实时定量聚合酶链反应(RT-qPCR)验证,结果与转录组测序数据一致。这些发现增进了我们对()耐缺氧性、复氧后恢复能力以及对缺氧反应的分子机制的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8417/11429291/14290a43cbbd/antioxidants-13-01063-g001.jpg

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