Institute of Hygienic and Environmental Medicinal Science, A Key Laboratory of Risk Assessment & Control for Environment & Food Safety, Tianjin 300050, China.
Toxicol Lett. 2010 Nov 10;199(1):69-79. doi: 10.1016/j.toxlet.2010.08.008. Epub 2010 Aug 21.
Pentachlorophenol (PCP) is a ubiquitous contaminant that has been shown to lead to hepatoxicity and is implicated in the incidence of liver tumors in human. A number of previous studies have described the toxic effects of PCP based on conventional toxicological indices. However, little evidence on protein levels is available at present. For further understanding of mechanisms of action and identifying the potential protein biomarkers for PCP exposure, two-dimensional electrophoresis coupled with mass spectrometry has been used to identify proteins differentially expressed in the livers of rare minnow (Gobiocypris rarus) following PCP exposure of 0.5, 5, 50 μg/L. After comparison of the protein profiles from treated and control groups, 39 protein spots were found altered in abundance (>2-fold) from male and female PCP-treated groups. Matrix-assisted laser desorption/ionization (MALDI) tandem time-of-flight mass spectrometry (TOF/MS) analysis allowed the unambiguous identification, and 18 protein spots were identified successfully, 12 proteins in females and 6 proteins in males, respectively. These proteins were involved in transport, metabolism, response to oxidative stress and other biological processes. Of these proteins, four differentially expressed mRNA encoding proteins underwent quantitative analysis by quantitative real-time PCR (QRT-PCR). The consistent and discrepant results between mRNA and protein levels suggested that complicated regulatory mechanisms of gene expression were implicated in the response to PCP exposure. In addition, marked gender differences in response to PCP have been described from the comparison of the male and female liver protein profiles.
五氯酚(PCP)是一种普遍存在的污染物,已被证明会导致肝毒性,并与人类肝癌的发病率有关。许多先前的研究已经基于传统的毒理学指标描述了 PCP 的毒性作用。然而,目前关于蛋白质水平的证据很少。为了进一步了解作用机制并确定 PCP 暴露的潜在蛋白质生物标志物,我们使用二维电泳结合质谱法鉴定了在 0.5、5、50μg/L PCP 暴露后稀有鲫(Gobiocypris rarus)肝脏中差异表达的蛋白质。比较处理组和对照组的蛋白质图谱后,从雄性和雌性 PCP 处理组中发现了 39 个蛋白质斑点的丰度发生了改变(>2 倍)。基质辅助激光解吸/电离(MALDI)串联飞行时间质谱(TOF/MS)分析允许明确鉴定,成功鉴定了 18 个蛋白质斑点,分别在雌性中有 12 个蛋白质斑点,雄性中有 6 个蛋白质斑点。这些蛋白质参与运输、代谢、对氧化应激的反应和其他生物过程。其中,四种差异表达的编码蛋白质的 mRNA 通过实时定量 PCR(QRT-PCR)进行了定量分析。mRNA 和蛋白质水平之间一致和不一致的结果表明,基因表达的复杂调控机制参与了对 PCP 暴露的反应。此外,从雄性和雌性肝脏蛋白质图谱的比较中描述了对 PCP 反应的明显性别差异。