Xiahui Ouyang, Xuelei Zhu, Lichao Wang, Xiangxiang Zheng, Juan Wang
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引用次数: 0
摘要
雌激素相关受体(Estrogen-related receptor, ERRs)是核受体(nuclear receptor, NR)家族中一大类孤儿受体。双酚A (BPA)被称为内分泌干扰物和异雌激素,与人类雌激素相关受体(ERRγ)有很高的亲和力。昆虫缺乏雌激素受体(ER),其ERR与ERRγ同源。本研究旨在探讨双酚a是否会影响蜂王卵巢孤儿核ERR的表达。采用聚合酶链反应(PCR)从蜜蜂的cDNA中扩增出amERR基因。收获amERR蛋白并用重组载体amERR/pET-32a纯化。采用RT-qPCR和Western blot (WB)技术检测对照组和处理组蜜蜂卵巢中amERR基因及蛋白表达水平。结果表明,3µg和10µg剂量均能抑制卵巢中amERR的表达,其中6H抑制作用最为显著。利用AutoDockVina模拟I-TASSER预测的amERR三级结构与BPA之间的分子对接。提示BPA可与amERR的配体结合袋(LBP)结合。因此,BPA可能通过结合阻止amERR的激活。结果表明,BPA可能会影响amERR基因的转录和蛋白表达水平。这可能与BPA与amERR的潜在特异性结合有关。
Effect of bisphenol A on the ovarian expressions of estrogen-related receptor gene and protein in queen honey bee (Apis mellifera)
Estrogen-related receptors (ERRs) belong to a large class of orphan receptors in the nuclear receptor (NR) family. Bisphenol A (BPA) is known as an endocrine disruptor and xenoestrogen with a high affinity for the human estrogen-related receptor (ERRγ). Insects lack estrogen receptors (ER), and their ERR is orthologous to ERRγ. This study was intended to examine whether BPA does not affect the ovarian expression of orphan nuclear ERR in queen honey bees (Apis mellifera L.). The amERR gene was amplified by polymerase chain reaction (PCR) from the cDNA of bees. The amERR protein was harvested and purified with the recombinant vector amERR/pET-32a. Using RT-qPCR and Western blot (WB), the levels of amERR gene and protein expression in the ovaries of control and treated bees were determined. The results indicated that both 3 µg and 10 µg doses suppressed the ovarian expression of amERR, with 6H inhibition being the most significant. Furthermore, AutoDockVina was utilized for simulating molecular docking between the tertiary structure of amERR predicted by I-TASSER and BPA. It hinted that BPA could conjugate with the ligand-binding pocket (LBP) of amERR. Thus, BPA might arrest amERR activation through binding. The results showed that BPA might impact the transcription and protein expression levels of the amERR gene. It may be correlated with the potential specific binding of BPA and amERR.
期刊介绍:
Apidologie is a peer-reviewed journal devoted to the biology of insects belonging to the superfamily Apoidea.
Its range of coverage includes behavior, ecology, pollination, genetics, physiology, systematics, toxicology and pathology. Also accepted are papers on the rearing, exploitation and practical use of Apoidea and their products, as far as they make a clear contribution to the understanding of bee biology.
Apidologie is an official publication of the Institut National de la Recherche Agronomique (INRA) and Deutscher Imkerbund E.V. (D.I.B.)