Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells
Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells
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DOI：
10.1006/taap.2002.9420
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发表时间：
2002-07-01
影响因子：
3.8
通讯作者：
van den Berg, M
中科院分区：
文献类型：
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作者：
Sanderson, JT;Boerma, J;van den Berg, M
在 H295R 细胞中筛选了各种已知或怀疑干扰类固醇激素功能的农药对芳香酶催化活性和 mRNA 表达的影响。二丁基氯化锡、三丁基氯化锡和三苯基氯化锡可降低芳香酶和乙氧基试卤灵 O-脱乙基酶的活性，浓度依赖性（1-300 nM；暴露 24 小时）。然而，这些减少仅发生在细胞毒性浓度下，线粒体 MTT 减少和细胞内中性红摄取的减少表明。有机锡在催化测定过程中不会引起直接抑制（1-1000 nM；暴露 1.5 小时）。 p,p'-DDT、o,p-DDT 和 o,p-DDE 也是如此，仅在细胞毒性浓度（大于或等于 10 muM；24 小时暴露）下才降低芳香酶活性。 1 和 10 muM 的 p,p'-DDE 对芳香酶活性或细胞活力没有影响。各种咪唑类杀菌剂都是芳香酶抑制剂。抑霉唑和咪鲜胺是有效的混合抑制剂（K-i/K-i' 分别为 0.04/0.3 和 0.02/0.3 muM），而丙环唑、苯醚甲环唑和戊菌唑是效力较弱的竞争性抑制剂（K-i 分别为 1.9、4.5 和 4.7 muM）。 Fenarimol、戊唑醇和己唑醇将芳香酶活性降低到接近细胞毒性浓度。正如先前对莠去津所显示的那样，乙烯菌唑啉可将芳香酶活性和 CYP19 mRNA 水平分别诱导约 2.5 倍和 1.5 倍。为了研究 H295R 细胞中芳香酶诱导的机制，检测了农药增加细胞内 cAMP 水平的能力。乙烯菌核利 (100 μM) 和莠去津 (30 μM) 使 cAMP 水平比对照增加约 1.5 倍。 Forskolin 和异丁基甲基黄嘌呤 (IBMX) 分别使 cAMP 水平增加 3 倍和 1.8 倍。乙烯菌核利、莠去津和IBMX诱导cAMP的时间-响应曲线和诱导芳香酶的浓度-响应曲线相似，表明这些农药诱导芳香酶的机制是通过抑制磷酸二酯酶活性来介导的。 (C) 2002 年爱思唯尔科学（美国）。
Various pesticides known or suspected to interfere with steroid hormone function were screened in H295R cells for effects on catalytic activity and mRNA expression of aromatase. Dibutyl-, tributyl-, and triphenyltin chloride decreased aromatase and ethoxyresorufin O-deethylase activities concentration dependently (1-300 nM; 24-h exposure). However, these decreases occurred only at cytotoxic concentrations, indicated by decreases in mitochondrial MTT reduction and intracellular neutral red uptake. The organotins did not cause direct inhibition during the catalytic assay (1-1000 nM; 1.5-h exposure). The same was true for p,p'-DDT, and o,p-DDT, and o,p-DDE, which decreased aromatase activity only at cytotoxic concentrations ( greater than or equal to10 muM; 24-h exposure). p,p'-DDE had no effect on aromatase activity or cell viability at 1 and 10 muM. Various imidazole-like fungicides were aromatase inhibitors. Imazalil and prochloraz were potent mixed inhibitors (K-i/K-i' = 0.04/0.3 and 0.02/0.3 muM, respectively), whereas propiconazole, difenoconazole, and penconazole were less potent competitive inhibitors (K-i = 1.9, 4.5, and 4.7 muM, respectively). Fenarimol, tebuconazole, and hexaconazole decreased aromatase activity close to cytotoxic concentrations. Vinclozolin, as was shown previously for atrazine, induced aromatase activity and CYP19 mRNA levels about 2.5- and 1.5-fold, respectively. To investigate the mechanism of aromatase induction in H295R cells, the ability of the pesticides to increase intracellular cAMP levels was examined. Vinclozolin (100 muM) and atrazine (30 muM) increased cAMP levels about 1.5-fold above control. Forskolin and isobutyl methylxanthine (IBMX) increased cAMP levels 3 and 1.8-fold, respectively. Time-response curves for cAMP induction and concentration-response curves for aromatase induction by vinclozolin, atrazine, and IBMX were similar, suggesting that the mechanism of aromatase induction by these pesticides is mediated through inhibition of phosphodiesterase activity. (C) 2002 Elsevier Science (USA).