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三嗪类除草剂阿特拉津对原代培养的睾丸间质细胞类固醇生成基因 mRNA 表达的影响及槲皮素的干预作用。

Effects of quercetin on mRNA expression of steroidogenesis genes in primary cultures of Leydig cells treated with atrazine.

机构信息

Department of Chemical Sciences, College of Natural Sciences, Redeemer's University, Redemption City, Ogun State, Nigeria.

出版信息

Toxicol In Vitro. 2013 Mar;27(2):700-7. doi: 10.1016/j.tiv.2012.11.005. Epub 2012 Nov 28.

DOI:10.1016/j.tiv.2012.11.005
PMID:23200735
Abstract

The effect of the phytoestrogen, quercetin (QT) on the reproductive toxicity of atrazine (ATZ) was explored in interstitial Leydig cells (ILCs). We measured the mRNA expressions of steroidogenesis genes: steroidogenic acute regulatory protein (StAR), 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome-P450 (CYP) 11A1, insulin-like factor 3 (INSL-3), CYP17A1, inhibin-α (INH-α), androgen receptor (AR), estrogen receptor-α (ER-α), and luteinising hormone receptor (LHR) in isolated ILCs by real-time-PCR after cultured cells were treated in vitro with ATZ (232 μM) and QT (50 μM). The mRNA expression of tested genes increased with ATZ treatment and was normalized by quercetin except AR and ER-α expression. Treatment of cells with QT alone (15-50 μM) caused a dose-dependent increase in AR and ER-α mRNA expression. We also found that QT (50 μM) increased the expressions of AR and ER-α in the presence of a sub-threshold level of cyclic-AMP at 1h culture period to the levels seen with maximal stimulation of cyclic-AMP. Furthermore, the expressions of tested genes were unaffected by cyclic-AMP at 6h when the stimulatory effects of ATZ on tested genes were sustained. These findings suggest that ATZ may stimulate the expression of tested steroidogenesis genes in ILCs via a mechanism independent of cyclic-AMP which was partially antagonize by QT.

摘要

探讨了植物雌激素槲皮素 (QT) 对阿特拉津 (ATZ) 生殖毒性的影响在间质莱迪希细胞 (ILCs) 中。我们通过实时 PCR 测量了类固醇生成基因的 mRNA 表达:类固醇急性调节蛋白 (StAR)、3β-羟类固醇脱氢酶 (3β-HSD)、细胞色素 P450 (CYP)11A1、胰岛素样因子 3 (INSL-3)、CYP17A1、抑制素-α (INH-α)、雄激素受体 (AR)、雌激素受体-α (ER-α) 和黄体生成素受体 (LHR) 在离体 ILCs 中,培养细胞用 ATZ(232 μM)和 QT(50 μM)体外处理后。用 ATZ 处理后,测试基因的 mRNA 表达增加,除 AR 和 ER-α 表达外,用槲皮素归一化。单独用 QT(15-50 μM)处理细胞会导致 AR 和 ER-α mRNA 表达呈剂量依赖性增加。我们还发现,QT(50 μM)在 1 小时培养期内,在环磷酸腺苷(cAMP)亚阈值水平下增加 AR 和 ER-α 的表达,使其达到最大 cAMP 刺激下的水平。此外,当 ATZ 对测试基因的刺激作用持续时,cAMP 在 6 小时时不会影响测试基因的表达。这些发现表明,ATZ 可能通过一种独立于 cAMP 的机制刺激 ILCs 中测试的类固醇生成基因的表达,QT 部分拮抗该机制。

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