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Effect of estradiol and nonylphenol on mRNA levels of vitellogenin II in the liver of chicken embryos

机译:雌二醇和壬基酚对鸡胚肝脏中卵黄蛋白原mRNA水平的影响

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摘要

Nonylphenol is one of the endocrine disrupting agents with estrogenic activity in some vertebrates. The present study was conducted to assess estrogenic activity of p-nonylphenol (NP) in chicken embryos by determining mRNA levels of liver vitellogenin II (VTG II). Fertile chicken eggs were incubated at 37.5 deg C using standard conditions. In the group 1, the eggs were treated with a single injection of either NP at doses of 50, 10, 1, 0.5, 0.2, 0.1, 0.01 and 0.001 mg/egg or estradiol-17 beta (E2)at doses of 1.0, 0.1 and 0.01 mg/egg in 50 mu l on day 16 of incubation. In the group 2, the eggs were treated with double injections of either NP or E2 on days 13 and 16 of incubation. In the control group the eggs were treated with the vehicle (propanediol (PD), 50 mu l/egg). On day 18 of incubation the liver was collected and total RNA was extracted. VTG II mRNA levels were determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. All the embryos were alive in the group of E2, PDor 0.01 and 0.001 mg/egg of NP. Only 17 percent (3/18) of embryos were alive in the group of NP at 10 mg/egg. No VTG II mRNA was detected in the control group, whereas distinct VTG II mRNA was revealed in the E2 treatment group where there was higher expression in the group 2 than in the group 1. Weak but distinct VTG II mRNA was detected in the NP treatment group. This study indicates that NP may have estrogenic activity in terms of liver VTG II mRNA assessed by RT-PCR assay in the chicken embryo.
机译:壬基酚是在某些脊椎动物中具有雌激素活性的内分泌干扰物之一。本研究旨在通过测定肝卵黄蛋白原II(VTG II)的mRNA水平来评估鸡胚中对壬基酚(NP)的雌激素活性。在标准条件下,将可育的鸡蛋在37.5摄氏度下孵育。在第1组中,以50、10、1、0.5、0.2、0.1、0.01和0.001 mg / egg的剂量单次注射NP或以1.0,在培养的第16天,每50毫升加入0.1和0.01毫克/蛋。在第2组中,在孵化的第13天和第16天,用NP或E2的双次注射处理卵。在对照组中,将卵用媒介物(丙二醇(PD),50μl/蛋)处理。在培养的第18天,收集肝脏并提取总RNA。 VTG II mRNA水平通过逆转录-聚合酶链反应(RT-PCR)分析确定。 E2,PDor 0.01和0.001 mg / egg NP组中的所有胚胎均活着。 NP组中只有10%/蛋的存活率为17%(3/18)。在对照组中未检测到VTG II mRNA,而在E2处理组中发现了明显的VTG II mRNA,其中在组2中的表达高于组1。在NP处理中检测到弱但独特的VTG II mRNA。组。这项研究表明,通过RT-PCR分析在鸡胚中,NP可能具有肝VTG II mRNA的雌激素活性。

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