孕期苯并a芘暴露对子代大鼠大脑皮质芳香烃受体表达及其与芳香烃受体核转运蛋白2相互作用的影响
Effects of benzoapyrene exposure during pregnancy on expression of aryl hydrocarbon receptor and its interaction with aryl hydrocarbon receptor nuclear transporter 2 in offspring rat cortex
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摘要:
背景 苯并
a 芘（BaP）作为生物异源性物质可以引起相关转录调节因子生理功能异常，但其具体机制尚不明确。目的 探究孕期BaP暴露对芳香烃受体（AhR）表达的影响以及AhR与转录因子芳香烃受体核转运蛋白2（ARNT2）的相互作用。
方法 取健康SD大鼠60只，以雌雄比例1：1的形式合笼。怀孕大鼠随机分为空白组、橄榄油组，以及10、20、40 mg·kg-1 BaP组，每组6只。受孕后17~19 d BaP组腹腔注射相应剂量BaP，橄榄油组注射等量橄榄油，空白组没有任何处理。取出生后1 d（PND1）和出生后7 d（PND7）的子鼠大脑皮质，使用Western blotting法和免疫共沉淀法分析不同时点子鼠AhR的表达及AhR与ARNT2相互作用的情况。
结果 Western blotting法检测结果显示：在PND1子鼠中，40mg·kg-1 BaP组皮质AhR蛋白相对表达水平（1.91±0.74）高于空白组（1.00±0.00）（
P < 0.05）；在PND7子鼠中，20、40 mg·kg-1 BaP组皮质AhR蛋白相对表达水平（1.92±0.56、1.77±0.62）高于空白组（1.00±0.00）（P < 0.05）。免疫共沉淀法检测结果显示：在PND1，ARNT2与AhR结合蛋白的相对表达量在10、20、40mg·kg-1 BaP组（1.74±0.27、2.55±0.40、1.26±0.24）均高于空白组（1.00±0.00）（P < 0.05）；在PND7，ARNT2与AhR结合蛋白的相对表达量在20、40 mg·kg-1 BaP组（1.75±0.59、1.83±0.49）均高于空白组（1.00±0.00）（P < 0.05）。结论 孕期BaP暴露会引起子鼠皮质内AhR表达水平升高，并且增强AhR与ARNT2的相互作用。
Abstract:Background Benzo
a pyrene (BaP), an xenobiotic compound, can induce dysfunction of related transcription regulators, but the underlying mechanism has not been clarified yet.Objective This study aims to investigate the effects of BaP exposure during pregnancy on aryl hydrocarbon receptor (AhR) expression and the interaction between AhR and aryl hydrocarbon receptor nuclear translocator 2 (ARNT2).
Methods Sixty healthy SD rats were selected and mated 1:1. The pregnant rats were divided into a blank group, an olive oil group, and 10, 20, and 40 mg·kg-1 BaP groups, with 6 rats in each group. At gestational day 17-19, the 30 pregnant rats were intraperitoneally injected with BaP, olive oil, or received no treatment as designed. The expression of AhR and its interaction with ARNT2 in the cortex of offspring rats on postnatal day 1 (PND1) and postnatal day 7 (PND7) were measured by Western blotting and Co-Immunoprecipitation assay.
Results According to the results of Western blotting: in the PND1 offspring cortex, the AhR relative expression level was increased in the 40 mg·kg-1 BaP group (1.91±0.74) compared with the blank group (1.00±0.00) (
P < 0.05); in the cortex of PND7 offspring, the AhR relative expression levels were increased in the 20 mg·kg-1 BaP group (1.92±0.56) and the 40 mg·kg-1 BaP group (1.77±0.62) (P < 0.05). The results of Co-Immunoprecipitation assay showed that, on PND1, the binding amounts of ARNT2 to AhR were increased in the 10, 20, and 40 mg·kg-1 BaP groups (1.74±0.27, 2.55±0.40, and 1.26±0.24, respectively) compared with the blank group (1.00±0.00) (P < 0.05); on PND7, the bindings amounts were increased in the 20 and 40mg·kg-1 BaP groups (1.75±0.59 and 1.83±0.49) compared with the blank group (1.00±0.00) (P < 0.05).Conclusion Prenatal BaP exposure can cause elevated offspring AhR expression and enhance AhR-ARNT2 interaction.
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