介绍一种LHRH调节LH分泌的离体模型——分散垂体细胞灌流技术  

CONTINUOUS PERFUSION OF DISPERSED ANTERIOR PITUITARY CELLS:A MODEL FOR LHRH REGULATION OF LH SECRETION

作　　者：屈智超 喻晶华 

机构地区：[1]四川省计划生育科学研究所生理室 [2]世界卫生组织人类生殖研究合作中心

出　　处：《生理学报》1990年第1期89-96,共8页Acta Physiologica Sinica

基　　金：WHO 人类生殖研究特别规划署资助

摘　　要：成年雄性 SD 大鼠断头后分离出垂体前叶(anterior pituitary,AP)。用胰蛋白酶消化和机械分散制备 AP 细胞(成活率大于95%)。分散的细胞悬液与生物凝胶混合后装上灌流柱,然后用 M199溶液连续灌流24h 以上。每间隔1~h 给予一次6min 的 LHRH 脉冲式刺激。细胞在此灌流过程中有一个稳定的基础 LH 分泌水平。LHRH 刺激能迅速引起 LH 分泌。对同一剂量 LHRH 的多次刺激可产生相同的 LH 脉冲。在一定的 LHRH 浓度范围内(1×10^(-10)_1×10^(-7)mol/L),LH 分泌与 LHRH 的剂量-效应曲线呈线性。实验结果表明,连续灌流分散的 AP 细胞的技术,优于单层细胞培养和组织块灌流等其他方法,是一种较为理想的研究LHRH 调节 LH 分泌机理的体外模型。The anterior pituitary(AP)tissues were removed from adult male Spraque-Dawley rats by decapitation.The dispersed cells with high viability(≥95%)were prepared using trypsin digestion and mechanical dissociation,and then mixedwith Bio-Gel P2,packed into the columns,and perfused continuously with M199medium for more than 24 hours.Different doses of LHRH were administeredby 6-min pulses at one hour intervals.A steady and detectable basal LH secretionwas present in all columns during the experiment.LHRH stimulation pulse couldinduce LH secretion rapidly,and repeated LHRH pulses of same dose producedstatistically equal LH pulses.The dose-response curve of LH secretion was linearwithin the range of 1×10^(-10) to 1×10^(-7)mol/L LHRH.These results indicate thatcontinuous perfusion system of dispersed AP cells which offers significant advantagesover other methodologies provides a very useful in vitro model for studying the me-chanisms on LHRH regulation of LH secretion.

关 键 词：分散垂体前叶细胞 灌流系统 离体模型 LHRH LH分泌 

分 类 号：Q575.12[生物学—生物化学] Q453