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Regulation of antioxidant systems in response to anoxia and reoxygenation in Rana sylvatica.
Comparative Biochemistry and Physiology B: Biochemistry & Molecular Biology ( IF 1.9 ) Pub Date : 2020-04-01 , DOI: 10.1016/j.cbpb.2020.110436 Aakriti Gupta 1 , Kenneth B Storey 1
Comparative Biochemistry and Physiology B: Biochemistry & Molecular Biology ( IF 1.9 ) Pub Date : 2020-04-01 , DOI: 10.1016/j.cbpb.2020.110436 Aakriti Gupta 1 , Kenneth B Storey 1
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The wood frog (Rana sylvatica) is a remarkable species. These frogs can endure prolonged oxygen deprivation as well as dehydration to ~60% of total body water lost and, combining these two abilities, they survive whole body freezing for weeks at a time during the winter. Episodes of anoxia/reoxygenation or freeze/thaw can trigger elevated production of reactive oxygen species (ROS) causing cellular damage, especially when oxygen is reintroduced during reoxygenation or thawing. To mitigate ROS damage, stress-responsive transcription factors such as the Octamer Binding Transcription factor (OCT4) and Nuclear factor (erythroid-derived 2)-like 2 transcription factor (Nrf2) were postulated to be involved in enhancing pro-survival pathways and antioxidant defenses. The present study used immunoblotting to analyze OCT4 and Nrf2 responses (and downstream factors under their control) to 24 h anoxia and 4 h reoxygenation in liver and skeletal muscle of wood frogs, with an emphasis on antioxidant systems. Surprisingly, no change was observed in relative total protein expression of either of the two transcription factors in liver. Furthermore, a significant decrease in total protein levels of OCT4 and Nrf2 occurred in skeletal muscle after 4 h recovery. However, essential cofactors of OCT4 and Nrf2 were significantly upregulated during anoxia and/or recovery. Downstream targets of the Nrf2-ARE pathway were evaluated, including glutathione-S-transferases (GSTs) and aldo-keto reductases (AKRs). Significant increases in GSTT1 and GSTP1 were observed in liver and muscle whereas AKRs showed a tissue specific response to both anoxia and recovery from anoxia. This study demonstrates activation of antioxidants as a cell protective mechanism against generation of reactive oxygen species during anoxia in wood frogs.
中文翻译：
林蛙中抗缺氧和复氧的抗氧化剂系统调节。
木蛙（Rana sylvatica）是一种非凡的物种。这些青蛙可以忍受长时间的缺氧以及脱水至约60％的体内总水分流失，结合这两种能力，它们在整个冬季一次可以冷冻数周。缺氧/复氧或冻结/解冻的发作可触发活性氧（ROS）产生增加，引起细胞损伤，特别是在复氧或解冻过程中重新引入氧气时。为了减轻ROS的损害，假定应激响应转录因子，例如Octamer结合转录因子（OCT4）和核因子（类胡萝卜素衍生的2）样2转录因子（Nrf2）参与增强生存途径和抗氧化剂防御。本研究使用免疫印迹法分析了木蛙的OCT4和Nrf2反应（以及受其控制的下游因素）对24 h缺氧和4 h复氧在木蛙的肝脏和骨骼肌中的反应，重点是抗氧化系统。令人惊讶地，在肝脏中两个转录因子中的任一个的相对总蛋白表达均未观察到变化。此外，恢复4 h后，骨骼肌中OCT4和Nrf2的总蛋白水平显着下降。但是，在缺氧和/或恢复过程中，OCT4和Nrf2的必需辅因子显着上调。评估了Nrf2-ARE途径的下游靶标，包括谷胱甘肽S-转移酶（GST）和醛基酮还原酶（AKR）。在肝脏和肌肉中观察到了GSTT1和GSTP1的显着增加，而AKRs显示了对缺氧和从缺氧恢复的组织特异性反应。这项研究表明，抗氧化剂的活化是防止青蛙在缺氧过程中产生活性氧的一种细胞保护机制。
更新日期：2020-04-01
中文翻译：
林蛙中抗缺氧和复氧的抗氧化剂系统调节。
木蛙（Rana sylvatica）是一种非凡的物种。这些青蛙可以忍受长时间的缺氧以及脱水至约60％的体内总水分流失，结合这两种能力，它们在整个冬季一次可以冷冻数周。缺氧/复氧或冻结/解冻的发作可触发活性氧（ROS）产生增加，引起细胞损伤，特别是在复氧或解冻过程中重新引入氧气时。为了减轻ROS的损害，假定应激响应转录因子，例如Octamer结合转录因子（OCT4）和核因子（类胡萝卜素衍生的2）样2转录因子（Nrf2）参与增强生存途径和抗氧化剂防御。本研究使用免疫印迹法分析了木蛙的OCT4和Nrf2反应（以及受其控制的下游因素）对24 h缺氧和4 h复氧在木蛙的肝脏和骨骼肌中的反应，重点是抗氧化系统。令人惊讶地，在肝脏中两个转录因子中的任一个的相对总蛋白表达均未观察到变化。此外，恢复4 h后，骨骼肌中OCT4和Nrf2的总蛋白水平显着下降。但是，在缺氧和/或恢复过程中，OCT4和Nrf2的必需辅因子显着上调。评估了Nrf2-ARE途径的下游靶标，包括谷胱甘肽S-转移酶（GST）和醛基酮还原酶（AKR）。在肝脏和肌肉中观察到了GSTT1和GSTP1的显着增加，而AKRs显示了对缺氧和从缺氧恢复的组织特异性反应。这项研究表明，抗氧化剂的活化是防止青蛙在缺氧过程中产生活性氧的一种细胞保护机制。
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