MS-222 induces biochemical and transcriptional changes related to oxidative stress, cell proliferation and apoptosis in zebrafish embryos
MS-222 induces biochemical and transcriptional changes related to oxidative stress, cell proliferation and apoptosis in zebrafish embryos
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DOI：
10.1016/j.cbpc.2020.108834
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发表时间：
2020-11-01
影响因子：
3.9
通讯作者：
Valentim, Ana M.
中科院分区：
文献类型：
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作者：
Felix, Luis M.;Luzio, Ana;Valentim, Ana M.
MS-222是鱼类中使用最广泛的麻醉剂，已被证明会对斑马鱼产生胚胎毒性效应。然而，潜在的分子效应仍然难以捉摸。本研究旨在通过评价生物化学和分子水平的变化来研究MS-222暴露对早期发育阶段的影响。在胚胎发育的3个阶段(256-细胞期、50%胚胎期和1-4体节期)分别用50、100和150 mg L-1MS-222作用20min，在8和26hpf两个时间点检测氧化应激、细胞增殖和细胞凋亡相关参数。在256细胞阶段暴露后，生化氧化还原平衡没有受到影响。与谷胱甘肽稳态相关的基因(gstpi和gclc)在8hpf时受到影响，而与细胞凋亡相关的基因(casp3a和casp6)和细胞增殖相关基因(增殖细胞核抗原)在26hpf时受到影响。过氧化氢酶活性在8hpf时呈倒U型反应。在50%胚胎期暴露后，与氧化应激相关的GCLC基因在8hpf上调，而gstpi下调，casp6随后上调，与谷胱甘肽过氧化物酶(GPx)活性下降和蛋白质羰基和casp3a非单调上调相一致。此外，MS-222处理的胚胎在26hpf时DCF染色减少。当暴露在1-4个体节阶段时，观察到类似的DCF染色模式。GPX活性也受到影响，而RT-qPCR显示caspase转录本呈剂量依赖性增加(casp3a、casp6和CASP9)。MS-222还上调了细胞增殖细胞核抗原的表达，同时改变了GCIC的表达。这些结果强调了MS-222对斑马鱼胚胎发育的影响，以及它对抗氧化剂、细胞增殖和细胞死亡系统的干扰机制尚不清楚；然而，结果表明Erk/Nrf2信号通路是一个目标候选。
MS-222, the most widely used anaesthetic in fish, has been shown to induce embryotoxic effects in zebrafish. However, the underlying molecular effects are still elusive. This study aimed to investigate the effects of MS-222 exposure during early developmental stages by evaluating biochemical and molecular changes. Embryos were exposed to 50, 100 or 150 mg L-1 MS-222 for 20 min at one of three developmental stages (256-cell, 50% epiboly, or 1-4 somite stage) and oxidative-stress, cell proliferation and apoptosis-related parameters were determined at two time-points (8 and 26 hpf). Following exposure during the 256-cell stage, the biochemical redox balance was not affected. The genes associated with glutathione homeostasis (gstpi and gclc) were affected at 8 hpf, while genes associated with apoptosis (casp3a and casp6) and cellular proliferation (pcna) were found affected at 26 hpf. An inverted U-shaped response was observed at 8 hpf for catalase activity. After exposure at the 50% epiboly stage, the gclc gene associated with oxidative stress was found upregulated at 8 hpf, while gstpi was downregulated and casp6 was upregulated later on, coinciding with a decrease in glutathione peroxidase (GPx) activity and a non-monotonic elevation of protein carbonyls and casp3a. Additionally, MS-222 treated embryos showed a decrease in DCF-staining at 26 hpf. When exposure was performed at the 1-4 somite stage, a similar DCF-staining pattern was observed. The activity of GPx was also affected whereas RT-qPCR showed that caspase transcripts were dose-dependently increased (casp3a, casp6 and casp9). The pcna mRNA levels were also found to be upregulated while gcic was changed by MS-222. These results highlight the impact of MS-222 on zebrafish embryo development and its interference with the antioxidant, cell proliferation and cellular death systems by mechanisms still to be explained; however, the outcomes point to the Erk/Nrf2 signalling pathway as a target candidate.