Evaluation of neuroactive steroid levels by liquid chromatography-tandem mass spectrometry in central and peripheral nervous system: Effect of diabetes
Evaluation of neuroactive steroid levels by liquid chromatography-tandem mass spectrometry in central and peripheral nervous system: Effect of diabetes
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DOI：
10.1016/j.neuint.2007.06.004
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发表时间：
2008-03-01
影响因子：
4.2
通讯作者：
Melcangi, Roberto C.
中科院分区：
文献类型：
--
作者：
Caruso, Donatella;Scurati, Samuele;Melcangi, Roberto C.
神经系统是神经活性类固醇的生理和保护作用的靶点。因此，在生理和病理条件下评估它们在神经结构中的水平是当务之急。为此，建立了液相色谱-串联质谱法（LC-MS/MS）对孕烯醇酮（PREG）、孕酮（PROG）、二氢孕酮（DHP）、四氢孕酮（THP）、睾酮(T)、二氢睾酮（DHT）、5 α -雄激素-3 α、17 β -二醇（3 α -二醇）、17 α -和17 β -雌二醇（17 α -e和17 β -e）进行鉴定和定量的方法。经验证后，应用该方法测定对照组和糖尿病大鼠中枢（即大脑皮层、小脑和脊髓）和外周（即臂神经）神经系统中神经活性类固醇的水平。在对照组中，只有臂神经检测到所有这些神经活性类固醇的水平。小脑17 α - e、大脑皮层17 α - e、17 β - e、DHP、THP、脊髓17 α - e、17 β - e、DHP均低于检测限。注射链脲佐菌素诱导的糖尿病强烈影响某些神经活性类固醇的水平。特别是小脑中PREG、PROG和T水平，大脑皮层中PROG、T和3 -二醇水平，脊髓中PROG、DHT和3 -二醇水平以及臂神经中PREG、DHP、THP、T、DHT和3 -二醇水平均显著降低。总之，本文报道的数据表明，LC-MS/MS方法可以以高灵敏度和特异性评估神经系统中的神经活性类固醇，糖尿病对其水平有强烈影响，为基于神经活性类固醇的新治疗工具提供了进一步的基础，旨在对抗糖尿病神经病变。(C) 2007 Elsevier Ltd.版权所有。
The nervous system is a target for physiological and protective effects of neuroactive steroids. Consequently, the assessment of their levels in nervous structures under physiological and pathological conditions is a top priority. To this aim, identification and quantification of pregnenolone (PREG), progesterone (PROG), dihydroprogesterone (DHP), tetrahydroprogesterone (THP), testosterone (T), dihydrotestosterone (DHT), 5 alpha-androstan-3 alpha, 17 beta-diol (3 alpha-diol), 17 alpha- and 17 beta-estradiol (17 alpha-E and 17 beta-E) by liquid chromatography and tandem mass spectrometry (LC-MS/MS) has been set up. After validation, this method was applied to determine the levels of neuroactive steroids in central (i.e., cerebral cortex, cerebellum and spinal cord) and peripheral (i.e., brachial nerve) nervous system of control and diabetic rats. In controls only the brachial nerve had detectable levels of all these neuroactive steroids. In contrast, 17 alpha-E in cerebellum, 17 alpha-E, 17 beta-E, DHP and THP in cerebral cortex, and 17 alpha-E, 17 beta-E and DHP in spinal cord were under the detection limit. Diabetes, induced by injection with streptozotocin, strongly affected the levels of some neuroactive steroids. In particular, the levels of PREG, PROG and T in cerebellum, of PROG, T and 3 alpha-diol in cerebral cortex, of PROG, DHT and 3 alpha-diol in spinal cord and of PREG, DHP, THP, T, DHT and 3 alpha-diol in brachial nerve were significantly decreased. In conclusion, the data here reported demonstrate that the LC-MS/MS method allows the assessment of neuroactive steroids in the nervous system with high sensitivity and specificity and that diabetes strongly affects their levels, providing a further basis for new therapeutic tools based on neuroactive steroids aimed at counteracting diabetic neuropathy. (C) 2007 Elsevier Ltd. All rights reserved.