title: Stress-induced changes of neurosteroid profiles in rat brain and plasma under immobilized condition.
abstract: 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.
authors.affiliation: Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Gyeonggi-do 15588, Korea.
authors.forename: Myeong Hyeon
authors.initials: MH
authors.lastname: Park
authors.affiliation: Department of Pharmacy, COMSATS Institute of Information Technology, Abbottabad, Pakistan.
authors.forename: Shaheed Ur
authors.initials: SU
authors.lastname: Rehman
authors.affiliation: Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Gyeonggi-do 15588, Korea.
authors.forename: In Sook
authors.initials: IS
authors.lastname: Kim
authors.affiliation: Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Gyeonggi-do 15588, Korea.
authors.forename: Min Sun
authors.initials: MS
authors.lastname: Choi
authors.affiliation: Institute of Pharmaceutical Science and Technology and College of Pharmacy, Hanyang University, Gyeonggi-do 15588, Korea. Electronic address: yoohh@hanyang.ac.kr.
authors.forename: Hye Hyun
authors.initials: HH
authors.lastname: Yoo
mesh_terms: D019342:Acetic Acid / Q000737:chemistry; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D000432:Methanol / Q000737:chemistry; D018377:Neurotransmitter Agents / Q000097:blood* / Q000737:chemistry / Q000378:metabolism*; D010949:Plasma / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D052616:Solid Phase Extraction / Q000379:methods
chemical_list: D018377:Neurotransmitter Agents; D019342:Acetic Acid; D000432:Methanol
keywords: Brain; Immobilization stress; LC–MS/MS; Neurosteroids; Plasma