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大黄素对团头鲂外周白细胞活性和Nrf2-Keap1通路中相关基因表达的影响
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摘要
本实验主要研究了团头鲂外周白细胞在不同浓度大黄素(0.04μg/ml、0.2μg/ml、1μg/ml、5μg/ml、25μg/ml)作用下细胞活性、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和Nrf2-Keap1通路中几种主要蛋白基因表达的影响。结果表明高浓度大黄素降低了白细胞活性,且呈剂量依赖。定量PCR结果显示低浓度大黄素诱导了SOD、CAT和核因子NF-E2相关因子-2(Nrf2)mRNA的基础表达水平,其中0.04μg/ml和0.2μg/ml大黄素显著促进了SOD mRNA表达水平,1μg/ml的大黄素显著促进了CAT和Nrf2 mRNA表达水平。而高浓度大黄素(5μg/ml和25μg/ml)显著降低了CAT mRNA表达水平。此外,Kelch样环氧氯丙烧相关蛋白-1(Keap1)和BTB-CNC异体同源蛋白-1(Bach1)mRNA表达水平随着大黄素浓度的增加而不断上升。本实验初步证实高浓度大黄素可降低白细胞活性,而白细胞在发生氧化应激时可能通过调节抗氧化酶和Nrf2-Keap1信号通路来诱导产生抗氧化应激反应。
This study aimed at investigating the effects of emodin exposure(0.04-25μg/ml) on cell viability andmRNA levels of antioxidative enzymes(SOD and CAT), and gene expressions of the Nrf2-Keap1 signaling molecules in the peripheral blood leukocytes of Megalobrama amblycephalafor 24 h. Results showed that cell viability effects of emodin were greatly reduced,and occurred in a dose-dependent manner. During low concentrations of exposure, mRNA levels of SOD in the cells treated with 0.04, 0.20 μg/ml, CATand 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. Furthermore, a sharp increase in Keap1 and Bach1 expression levels were observed a dose-dependent manner.
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