Real-time in vivo imaging reveals localised Nrf2 stress responses associated with direct and metabolism-dependent drug toxicity.
Real-time in vivo imaging reveals localised Nrf2 stress responses associated with direct and metabolism-dependent drug toxicity.
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DOI：
10.1038/s41598-017-16491-2
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发表时间：
2017-11-22
影响因子：
4.6
通讯作者：
Copple IM
中科院分区：
文献类型：
--
作者：
Forootan SS;Mutter FE;Kipar A;Iwawaki T;Francis B;Goldring CE;Park BK;Copple IM
转录因子Nrf2协调对化学和氧化应激的适应性反应，其特征是细胞保护靶基因的表达上调。为了了解Nrf2作为药物应激标志物的机制相关性，了解这种适应性反应是否真正局限于器官特异性药物毒性是很重要的。在这里，我们通过在给予代谢依赖肝毒素对乙酰氨基酚(APAP)或直接肾毒素顺铂后，对转基因NRF2-荧光素酶(NRF2-Luc)报告小鼠进行实时生物发光成像来解决这一知识差距。我们在体内和体外检测到肝脏(APAP)和肾脏(顺铂)的局部生物发光，而qPCR、Taqman低密度阵列和免疫印迹分析进一步表明这些组织中几个内源性Nrf2调节的基因/蛋白的表达水平增加，包括血红素加氧酶1(Hmox1)。与APAP对肝脏和顺铂对肾脏的毒性作用一致，免疫组织化学分析显示荧光素酶和Hmox1在小叶中心肝细胞和肾小管上皮细胞中的表达分别升高。细胞色素P450抑制剂氨基苯并三氮唑可改善APAP的肝毒性和局部的Nrf2-Luc反应，这与反应代谢物的形成在APAP诱导的化学应激中的作用是一致的。总之，这些发现表明，Nrf2可以反映与相关毒理学机制相关的高度局部化的细胞扰动。
The transcription factor Nrf2 coordinates an adaptive response to chemical and oxidative stress characterised by the upregulated expression of cytoprotective target genes. In order to understand the mechanistic relevance of Nrf2 as a marker of drug-induced stress it is important to know if this adaptive response is truly localised in the context of organ-specific drug toxicity. Here, we address this knowledge gap through real-time bioluminescence imaging of transgenic Nrf2-luciferase (Nrf2-luc) reporter mice following administration of the metabolism-dependent hepatotoxin acetaminophen (APAP) or the direct nephrotoxin cisplatin. We detected localised bioluminescence in the liver (APAP) and kidneys (cisplatin) in vivo and ex vivo, whilst qPCR, Taqman low-density array and immunoblot analysis of these tissues further revealed increases in the expression level of several endogenous Nrf2-regulated genes/proteins, including heme oxygenase 1 (Hmox1). Consistent with the toxic effects of APAP in the liver and cisplatin in the kidney, immunohistochemical analysis revealed the elevated expression of luciferase and Hmox1 in centrilobular hepatocytes and in tubular epithelial cells, respectively. In keeping with the role of reactive metabolite formation in APAP-induced chemical stress, both the hepatotoxicity and localised Nrf2-luc response were ameliorated by the cytochrome P450 inhibitor aminobenzotriazole. Together, these findings show that Nrf2 can reflect highly-localised cellular perturbations associated with relevant toxicological mechanisms.
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影响因子：
13.5
作者：
Lundback, Peter;Lea, Jonathan D.;Sowinska, Agnieszka;Ottosson, Lars;Furst, Camilla Melin;Steen, Johanna;Aulin, Cecilia;Clarke, Joanna I.;Kipar, Anja;Klevenvall, Lena;Yang, Huan;Palmblad, Karin;Park, B. Kevin;Tracey, Kevin J.;Blom, Anna M.;Andersson, Ulf;Antoine, Daniel J.;Harris, Helena Erlandsson
通讯作者：
Harris, Helena Erlandsson
影响因子：
3.9
作者：
Mutter FE;Park BK;Copple IM
通讯作者：
Copple IM
影响因子：
46.9
作者：
通讯作者：
--
影响因子：
4.6
作者：
Clarke, Jessica L.;Murray, James B.;Copple, Ian M.
通讯作者：
Copple, Ian M.
影响因子：
5.7
作者：
Yates, Melinda S.;Tauchi, Masafumi;Kensler, Thomas W.
通讯作者：
Kensler, Thomas W.