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首页> 外文期刊>The Journal of Experimental Biology >Inductive transcription and protective role of fish heme oxygenase-1 under hypoxic stress.
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Inductive transcription and protective role of fish heme oxygenase-1 under hypoxic stress.

机译：低氧胁迫下鱼血红素加氧酶-1的诱导转录和保护作用。

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Heme oxygenase-1 is the rate-limiting enzyme in the degradation of heme into biliverdin, carbon monoxide and free divalent iron. In this study, we cloned heme oxygenase isoform 1 (CaHO-1) from a hypoxia-tolerant teleost fish Carassius auratus. The full-length cDNA of CaHO-1 is 1247 bp and encodes a protein of 272 amino acids. RT-PCR and real-time PCR analysis indicated that CaHO-1 was predominantly transcribed in posterior kidney, head kidney, gill and intestine, and induction of gene transcription was observed predominantly in posterior kidney under hypoxic stress. Moreover, the hypoxia-induced transcription was confirmed in goldfish larvae and in in vitro cultured CAB cells. Fluorescence of the HO-1-GFP fusion protein revealed a cytoplasmic and plasma membrane localization, which was consistent with the putative transmembrane structure. Subsequently, we established a stably transfected CAB/pcDNA3.1-HO-1 cell line and a control CAB/pcDNA3.1 cell line, and found that the number of dead cells was obviously reduced in the pcDNA3.1-HO-1-transfected group following 4 days of hypoxic (1% O(2)) treatment in comparison with numerous detached dead cells in the control pcDNA3.1-transfected cells. Furthermore, a significant cell viability difference between the two kinds of transfected cells during hypoxia-reoxygenation was revealed. Therefore, the data suggest that fish HO-1 might play a significant protective role in cells in response to hypoxic stress.
机译：血红素加氧酶-1是将血红素降解为胆绿素，一氧化碳和游离二价铁的限速酶。在这项研究中，我们从耐缺氧的硬骨鱼Car鱼（Carassius auratus）中克隆了血红素加氧酶同工型1（CaHO-1）。 CaHO-1的全长cDNA为1247 bp，编码272个氨基酸的蛋白质。 RT-PCR和实时PCR分析表明CaHO-1主要在后肾，头肾、,和肠中转录，并且在低氧胁迫下在后肾中主要观察到基因转录的诱导。此外，在金鱼幼虫和体外培养的CAB细胞中证实了低氧诱导的转录。 HO-1-GFP融合蛋白的荧光显示胞质和质膜的定位，这与推定的跨膜结构一致。随后，我们建立了稳定转染的CAB / pcDNA3.1-HO-1细胞系和对照CAB / pcDNA3.1细胞系，发现pcDNA3.1-HO-1-中的死细胞数量明显减少。转染组经过4天的低氧（1％O（2））处理后，与对照pcDNA3.1转染的细胞中大量脱落的死细胞相比。此外，揭示了在低氧-复氧期间两种转染细胞之间的显着细胞活力差异。因此，数据表明鱼HO-1在低氧胁迫下可能在细胞中起重要的保护作用。

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