Department of Chemistry, College of Natural Sciences, Hanyang University , Seoul 133-791, South Korea.
Environ Sci Technol. 2013 Dec 3;47(23):13804-12. doi: 10.1021/es403269v. Epub 2013 Nov 14.
Cytochrome P450 (CYP) proteins are involved in the first line of detoxification mechanism against diverse polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P). In aquatic invertebrates, there is still a lack of knowledge on the CYP genes involved in the molecular response to B[a]P exposure due to limited gene information. In this study, we cloned the entire 25 CYP genes in the monogonont rotifer Brachionus koreanus with the aid of next generation sequencing (NGS) technologies and analyzed their transcript profiles with a real-time RT-PCR array to better understand B[a]P-triggered molecular response over different time courses. As a result, B[a]P exposure induced CYP2/3-involved detoxification mechanisms and defensome, including phase II detoxification and antioxidant systems with a modulation of the chaperone heat shock protein (hsp) expression but did not change expression of other CYP clans in B. koreanus . Therefore, we found that B[a]P induced a strong detoxification mechanism to overcome detrimental effects of B[a]P associated with B[a]P-induced growth retardation as a trade-off in fitness costs. Also, this approach revealed that the entire CYP profiling can be a way of providing a better understanding on the mode of action of B[a]P in B. koreanus with respect to molecular defense metabolism.
细胞色素 P450(CYP)蛋白参与了多种多环芳烃(PAHs)的第一线解毒机制,包括苯并[a]芘(B[a]P)。在水生无脊椎动物中,由于基因信息有限,对于参与 B[a]P 暴露的分子反应的 CYP 基因仍然知之甚少。在这项研究中,我们借助下一代测序(NGS)技术在单巢轮虫 Brachionus koreanus 中克隆了 25 个完整的 CYP 基因,并通过实时 RT-PCR 阵列分析了它们的转录谱,以更好地了解不同时间过程中 B[a]P 引发的分子反应。结果表明,B[a]P 暴露诱导了 CYP2/3 参与的解毒机制和防御体,包括 II 相解毒和抗氧化系统,同时调节伴侣热休克蛋白(hsp)的表达,但不会改变 B. koreanus 中其他 CYP 族的表达。因此,我们发现 B[a]P 诱导了一种强烈的解毒机制,以克服与 B[a]P 相关的生长迟缓的不利影响,这是一种适应成本的权衡。此外,这种方法表明,整个 CYP 分析可以提供一种更好的理解方式,即 B[a]P 在 B. koreanus 中的作用模式,涉及分子防御代谢。