Computational analysis of liquid chromatography-tandem mass spectrometric steroid profiling in NCI H295R cells following angiotensin II, forskolin and abiraterone treatment
Computational analysis of liquid chromatography-tandem mass spectrometric steroid profiling in NCI H295R cells following angiotensin II, forskolin and abiraterone treatment
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DOI：
10.1016/j.jsbmb.2015.09.038
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发表时间：
2016-01-01
影响因子：
4.1
通讯作者：
Eisenhofer, Graeme
中科院分区：
文献类型：
--
作者：
Mangelis, Anastasios;Dieterich, Peter;Eisenhofer, Graeme
调节过多生理功能的肾上腺类固醇激素通过严格控制的途径产生。这些途径的调查，实验数据的基础上，可以促进计算代谢率的变化计算建模。因此，我们使用了一个模型系统，质量平衡和质量反应方程的基础上，在人肾上腺皮质衍生的NCI H295 R细胞中，肾上腺类固醇生成的动力学评价。为此，通过液相色谱-串联质谱法测量了一组10种类固醇。与血管紧张素II，毛喉素和阿比特龙孵育后，测定细胞孵育浓度的类固醇，包括皮质醇，醛固酮，脱氢表雄酮及其前体的时间依赖性变化。基于实验数据，采用加权最小二乘法拟合模型参数。时间依赖性血管紧张素II和毛喉素诱导的变化，观察到孵育浓度的前体类固醇的峰值之前，最大增加的醛固酮和皮质醇。用阿比特龙抑制17-α-羟化酶/17，20-裂解酶导致上游前体类固醇增加和下游产物减少。推导出的模型参数，包括酶促过程的速率常数，适当量化了多个转化步骤中代谢途径的观察和预期变化。我们的数据表明单一时间点测量的局限性和评估途径动力学在肾上腺皮质细胞系类固醇生成研究中的重要性。我们的分析提供了一个框架，肾上腺皮质细胞培养系统中的类固醇合成的评价，并表明，计算建模衍生的动力学参数的估计是一个有效的工具，用于描述相关的代谢途径的扰动。(C)2015爱思唯尔有限公司版权所有。
Adrenal steroid hormones, which regulate a plethora of physiological functions, are produced via tightly controlled pathways. Investigations of these pathways, based on experimental data, can be facilitated by computational modeling for calculations of metabolic rate alterations. We therefore used a model system, based on mass balance and mass reaction equations, to kinetically evaluate adrenal steroidogenesis in human adrenal cortex-derived NCI H295R cells. For this purpose a panel of 10 steroids was measured by liquid chromatographic-tandem mass spectrometry. Time-dependent changes in cell incubate concentrations of steroids including cortisol, aldosterone, dehydroepiandrosterone and their precursors were measured after incubation with angiotensin II, forskolin and abiraterone. Model parameters were estimated based on experimental data using weighted least square fitting. Time-dependent angiotensin II- and forskolin-induced changes were observed for incubate concentrations of precursor steroids with peaks that preceded maximal increases in aldosterone and cortisol. Inhibition of 17-alpha-hydroxylase/17,20-lyase with abiraterone resulted in increases in upstream precursor steroids and decreases in downstream products. Derived model parameters, including rate constants of enzymatic processes, appropriately quantified observed and expected changes in metabolic pathways at multiple conversion steps. Our data demonstrate limitations of single time point measurements and the importance of assessing pathway dynamics in studies of adrenal cortical cell line steroidogenesis. Our analysis provides a framework for evaluation of steroidogenesis in adrenal cortical cell culture systems and demonstrates that computational modeling-derived estimates of kinetic parameters are an effective tool for describing perturbations in associated metabolic pathways. (C) 2015 Elsevier Ltd. All rights reserved.