Park Myeong Hyeon, Rehman Shaheed Ur, Kim In Sook, Choi Min Sun, Yoo Hye Hyun
Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Gyeonggi-do 15588, Korea.
Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad, Pakistan.
J Pharm Biomed Anal. 2017 May 10;138:92-99. doi: 10.1016/j.jpba.2017.02.007. Epub 2017 Feb 3.
In this study, various neurosteroids in brain and plasma were simultaneously determined using liquid chromatography-tandem mass spectrometry and their profile changes in a stress-induced rats were investigated. The investigated neurosteroids are as follows: progesterone (P4), 5α-dihydroprogesterone (5α-DHP), 5β-dihydroprogesterone, estrone, androstenedione (AE), cortisol, cortisone, corticosterone (CORT), dehydroepiandrosterone (DHEA), pregnanolone (3α,5β-THP), allopregnanolone (ALLO), 11-deoxycorticosterone (DOC), 11-deoxycortisol, pregnenolone (PREG), and 5α/5β-tetrahydrodeoxycorticosterone (5α/5β-THDOC). Brain and plasma samples were processed using solid-phase extraction with methanol and acetic acid (99:1), and derivatized with a hydroxylamine reagent. Separation was achieved within 13min at a flow rate of 0.4mL/min with a C18 column (3.0×50mm, 2.7μm). The triple quadrupole mass spectrometer was operated in the positive electrospray ionization mode. Using this method, the neurosteroid level variation was quantitated and investigated in the brain and plasma upon immobilization stress in rats. As a result, AE, CORT, DOC, P4, 5α-DHP, 5α/5β-THDOC, DHEA, 3α,5β-THP, ALLO, and PREG levels were significantly altered in both the brain and plasma samples when stress was induced. These findings demonstrated that stress leads to the alteration of the GABAergic neurosteroid profile. The present results will be helpful for furthering an understanding of the role of neurosteroids in stressed conditions.
在本研究中，使用液相色谱 - 串联质谱法同时测定大脑和血浆中的各种神经甾体，并研究其在应激诱导大鼠中的谱变化。所研究的神经甾体如下：孕酮（P4）、5α - 二氢孕酮（5α - DHP）、5β - 二氢孕酮、雌酮、雄烯二酮（AE）、皮质醇、可的松、皮质酮（CORT）、脱氢表雄酮（DHEA）、孕烷醇酮（3α,5β - THP）、别孕烷醇酮（ALLO）、11 - 脱氧皮质酮（DOC）、11 - 脱氧皮质醇、孕烯醇酮（PREG）以及5α/5β - 四氢脱氧皮质酮（5α/5β - THDOC）。大脑和血浆样本用甲醇和乙酸（99:1）进行固相萃取处理，并用羟胺试剂衍生化。使用C18柱（3.0×50mm，2.7μm），以0.4mL/min的流速在13分钟内实现分离。三重四极杆质谱仪在正电喷雾电离模式下运行。使用该方法，对大鼠固定应激后大脑和血浆中的神经甾体水平变化进行了定量和研究。结果表明，当诱导应激时，大脑和血浆样本中的AE、CORT、DOC、P4、5α - DHP、5α/5β - THDOC、DHEA、3α,5β - THP、ALLO和PREG水平均发生显著变化。这些发现表明应激导致GABA能神经甾体谱的改变。本研究结果将有助于进一步理解神经甾体在应激条件下的作用。
