雌性大鼠血清和海马雌二醇水平及海马孕烯醇酮水平的动态变化  

Dynamic Changes of Estradiol and Pregnenolone Levels in Serum and Hippocampus of Female Rats during Different Estrous Cycle

作　　者：宗煜[1,2] 马昱[1] 林豪杰[1] 汪昕[1] 范薇[1] 

机构地区：[1]复旦大学附属中山医院神经内科,上海200032 [2]青岛市立医院神经内科,山东青岛266011

出　　处：《神经损伤与功能重建》2009年第4期252-256,共5页Neural Injury and Functional Reconstruction

摘　　要：目的:建立检测大鼠海马组织中雌二醇(E2)水平和孕烯醇酮(PREG)水平的方法,观察正常雌性大鼠血清及海马E2水平和海马PREG水平的动态变化。方法:24只健康成年雌性SD大鼠分为发情前期组、发情期组、发情后期组和发情间期组各6只;取血清及海马组织,用有机溶剂(乙酸乙酯/正己烷=3/2)萃取甾体激素后,评估高效液相色谱(HPLC)分离、纯化E2和PREG的性能指标(包括分离效果、日内/日间变异率、回收率和总回收率),采用HPLC分离、纯化萃取物中的E2和PREG;应用放射免疫法(RIA)检测E2,应用酶联免疫吸附分析(ELISA)检测PREG。结果:HPLC分离E2和PREG的保留时间日内变异率分别为0.3%和0.4%,日间变异率分别为0.6%和0.4%;不同浓度的E2和PREG的回收率与总回收率差异无统计学意义(P>0.05)。雌性大鼠血清和海马E2水平分别在发情前期和发情期最高,与其它各期相比差异有统计学意义(P<0.05),各期大鼠海马E2水平(1.3～1.9fmoL.mg-1)均高于血清E2峰值水平(0.69fmoL.mg-1)(P<0.001)。各期大鼠海马PREG水平差异无统计学意义(P>0.05)。结论:本实验检测脑内E2和PREG的方法灵敏且可靠。正常雌性大鼠海马组织中的E2可能主要由自身局部合成,并且有可能是通过芳香化酶的作用由睾酮直接转化而来。Objective：To develop a method for the determination of estradiol （E2） and pregnenolone （PREG） levels in serum and hippocampus of female rats ; and to observe the dynamic changes of E2 in serum and hippocampus and hippocampal PREG levels in female rats during different estrous cycle. Methods：The female SD rats were divided into proestrus, estrus, metestrus and diestrus groups （each group： n=6）. The steroid hormones were ex- tracted from serums and hippocampal tissues by an organic solvent, and then the E2 and PREG were separated and purified by high performance liquid chromatography （HPLC）. The levels of E2 and PREG were measured by Radio- immunoassay （RIA） and Enzyme linked immunosorbent assay （ELISA） respectively. Results：The separation of E2 and PREG by HPLC showed good reproducibility [the coefficients of variation （CVs） ]. Within each day （n=6） retention time was 0.3% and 0.4%, and the CVs between-day were 0.6% and 0.4%, respectively. Moreover, there was no significant differences in the recovery of E2 and PREG at different concentrations. The levels of E2 in serum and hippocampus were significantly higher in the proestrus and estrus groups than those in the other groups. The concentration of Ez （1. 3 - 1. 9 fmoL. mg -1） in the hippocampus was much higher than those in the serum （0. 69 fmoL. mg -1 ）. PREG levels in hippocampus showed no significant difference during the estrous cycle. Con- clusion;The established method to detect E2 and PREG of brain tissue was sensitive and reliable. E2 level in rat hip- pocampus is higher than that in serum, and there were no change of PREG content during estrus cycle. Our results suggest that the E2 in hippocampus of female rat is mainly endogenous.

关 键 词：雌二醇 孕烯醇酮 血清 海马 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学] R742.1[医药卫生—临床医学]