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大型溞和颤蚓中的多异生物质抗性外排活性。

Multixenobiotic resistance efflux activity in Daphnia magna and Lumbriculus variegatus.

作者信息

Vehniäinen Eeva-Riikka, Kukkonen Jussi V K

机构信息

University of Jyväskylä, Department of Biological and Environmental Science, P.O. Box 35, FI-40014 University of Jyväskylä, Finland.

出版信息

Chemosphere. 2015 Apr;124:143-9. doi: 10.1016/j.chemosphere.2014.12.014. Epub 2014 Dec 27.

Abstract

Multixenobiotic resistance is a phenomenon in which ATP-binding cassette (ABC) family proteins transfer harmful compounds out of cells. Daphnia magna and Lumbriculus variegatus are model species in aquatic ecotoxicology, but the presence and activity of ABC proteins have not been well described in these species. The aim of this work was to study the presence, activity, and inhibition of ABC transport proteins in D. magna and L. variegatus. The presence of abcb1 and abcc transcripts in 8-9-day-old D. magna was investigated by qRT-PCR. The activity of MXR in D. magna and L. variegatus was explored by influx of the fluorescent ABC protein substrates rhodamine B and calcein-AM, with and without the model inhibitors verapamil (unspecific ABC inhibitor), reversin 205 (ABCB1 inhibitor) and MK571 (ABCC inhibitor). Juvenile D. magna possessed all examined abcb and abcc transcripts, but only reversin 205 inhibited MXR activity. The MXR activity in L. variegatus was inhibited by MK571, and to a lesser extent by verapamil, whereas reversin 205 seemed to stimulate the transport activity. Whereas calcein-AM worked better as an MXR substrate in D. magna, rhodamine B was a better substrate for L. variegatus MXR activity measurements. This is the first report on MXR activity in the order Lumbriculida, subclass Oligochaeta, and class Clitellata.

摘要

多异种生物抗性是一种ATP结合盒(ABC)家族蛋白将有害化合物转运出细胞的现象。大型溞和颤蚓是水生生态毒理学中的模式物种,但这些物种中ABC蛋白的存在和活性尚未得到充分描述。这项工作的目的是研究大型溞和颤蚓中ABC转运蛋白的存在、活性及抑制情况。通过qRT-PCR研究了8 - 9日龄大型溞中abcb1和abcc转录本的存在情况。利用荧光ABC蛋白底物罗丹明B和钙黄绿素-AM的内流情况,在有和没有模型抑制剂维拉帕米(非特异性ABC抑制剂)、瑞维菌素205(ABCB1抑制剂)和MK571(ABCC抑制剂)的情况下,探究大型溞和颤蚓中多异种生物抗性(MXR)的活性。幼年大型溞拥有所有检测的abcb和abcc转录本,但只有瑞维菌素205抑制MXR活性。MK571抑制了颤蚓中的MXR活性,维拉帕米的抑制作用较小,而瑞维菌素205似乎刺激了转运活性。在大型溞中,钙黄绿素-AM作为MXR底物效果更好,而在测量颤蚓的MXR活性时,罗丹明B是更好的底物。这是关于寡毛纲颤蚓目和环带纲中MXR活性的首次报道。

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