Zhao Zhenxin, Xie Jun, Liu Bo, Ge Xianping, Song Changyou, Ren Mingchun, Zhou Qunlan, Miao Linghong, Zhang Huimin, Shan Fan, Yang Zhenfei
Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China.
Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China; Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Fish Shellfish Immunol. 2017 Mar;62:75-85. doi: 10.1016/j.fsi.2017.01.006. Epub 2017 Jan 5.
We determined the effects of emodin on the cell viability, respiratory burst activity, mRNA levels of antioxidative enzymes (Cu-Zn SOD, CAT and NOX2), and gene expressions of the Nrf2-Keap1 signaling molecules in the peripheral blood leukocytes of blunt snout bream. Triplicate groups of cultured cells were treated with different concentrations of emodin (0.04-25 μg/ml) for 24 h. Results showed that the emodin caused a dramatic loss in cell viability, and occurred in a dose-dependent manner. Emodin exposure (1-25 μg/ml) were significantly induced the ROS generation compared to the control. The respiratory burst and NADPH oxidase activities were significantly induced at a concentration of 0.20 μg/ml, and inhibited at 25 μg/ml. Besides, mRNA levels of antioxidant enzyme genes were dramatically regulated by emodin exposure for 24 h. During low concentrations of exposure, mRNA levels of Cu-Zn SOD in the cells treated with 0.04, 0.20 μg/ml, CAT, NOX2 and Nrf2 in the cells treated with 1 μg/ml were sharply increased, respectively. Whereas, high concentrations were dramatically down-regulated the gene expressions of CAT in the cells treated with 5, 25 μg/ml and NOX2 in the cells treated with 25 μg/ml. Furthermore, sharp increase in Keap1and Bach1 expression levels were observed a dose-dependent manner. In conclusion, this study demonstrated that emodin could induce antioxidant defenses which were involved in cytotoxic activities, respiratory burst and the transcriptional regulation levels of antioxidant enzymes and Nrf2-Keap1 signaling molecules.
我们测定了大黄素对团头鲂外周血白细胞的细胞活力、呼吸爆发活性、抗氧化酶（铜锌超氧化物歧化酶、过氧化氢酶和NADPH氧化酶2）的mRNA水平以及Nrf2-Keap1信号分子基因表达的影响。将培养细胞分成三组，用不同浓度（0.04 - 25μg/ml）的大黄素处理24小时。结果表明，大黄素导致细胞活力显著丧失，且呈剂量依赖性。与对照组相比，大黄素暴露（1 - 25μg/ml）显著诱导了活性氧的产生。在浓度为0.20μg/ml时，呼吸爆发和NADPH氧化酶活性显著诱导，而在25μg/ml时受到抑制。此外，大黄素暴露24小时显著调节了抗氧化酶基因的mRNA水平。在低浓度暴露时，用0.04、0.20μg/ml处理的细胞中铜锌超氧化物歧化酶的mRNA水平，以及用1μg/ml处理的细胞中过氧化氢酶、NADPH氧化酶2和Nrf2的mRNA水平分别急剧增加。然而，高浓度时，用5、25μg/ml处理的细胞中过氧化氢酶的基因表达，以及用25μg/ml处理的细胞中NADPH氧化酶2的基因表达显著下调。此外，观察到Keap1和Bach1表达水平呈剂量依赖性急剧增加。总之，本研究表明大黄素可诱导抗氧化防御，这涉及细胞毒性活性、呼吸爆发以及抗氧化酶和Nrf2-Keap1信号分子的转录调控水平。
