胎兔动脉导管平滑肌细胞氧敏感性钾通道mRNA的表达  

Expression of Kv1.5 Gene in Smooth Muscle Cells of Ductus Arteriosus in Fetal Rabbit

作　　者：余忠琴[1] 李涛[1] 

机构地区：[1]郧阳医学院附属太和医院儿童医疗中心二病区,湖北十堰442000

出　　处：《武汉大学学报（医学版）》2009年第1期21-23,51,I0002,共5页Medical Journal of Wuhan University

摘　　要：目的:建立胎兔动脉导管平滑肌细胞的体外培养方法,并检测其细胞膜氧敏感性钾通道亚型(Kv1.5)mRNA的表达。方法:采用组织块改良贴壁法(玻片压迫组织块贴壁方法)进行胎兔动脉导管平滑肌细胞的体外原代培养,并行免疫组织化学法对培养的细胞进行鉴定,用RT-PCR的方法检测其细胞膜氧敏感性钾通道亚型(Kv1.5)的表达。结果:经10-14 d培养后,培养皿底及盖玻片上铺满梭状细胞,免疫组织化学法鉴定确认为胎兔动脉导管平滑肌细胞,其细胞膜存在Kv1.5基因表达。结论:植块改良贴壁法可以成功的进行胎兔动脉导管平滑肌细胞的体外培养,且本法简便、可靠,短时间内可以获得大量细胞,从而为动脉导管未闭发病机制的研究奠定了实验基础。同时,动脉导管平滑肌细胞膜上存在氧敏感性钾通道mRNA的表达,从而为从氧敏感性钾通道层面来研究动脉导管未闭的发生机制提供了实验基础。Objective： To set up a method of cultivating fetal rabbit ductus arteriosus smooth muscle cells in vitro and to investigate the expression of Kvl. 5 mRNA in the cultured ductus arteriosus smooth muscle cells. Methods： Fetal rabbit smooth muscle ceils from ductus arteriosus were primary cultured by modified explant attached method that tissues were pressed with glass slices in vitro, and the cultured cells were identified using immunohistochemistry. The RT-PCR assay was used to investigate the mRNA expression of Kv 1.5. Results： After 10-14 days, spindle cells spreaded over the bottom of culture capsule and the glass slice, and were confirmed as smooth muscle cells of ductus arteriosus by immunohistochemistry. The Kv1. 5 gene was expressed in cultured ductus arteriosus smooth muscle cells. Conclusion： Smooth muscle cells of ductus arteriosus could be culture by modified explant attached method in vitro and proliferated well, and Kv1. 5 mRNA expression could be detected in these cells. It provides a useful and reliable method of experiment foundation for investigating the mechanism of patent dusctus arteriosus.

关 键 词：氧敏感性钾通道(Kv1.5)mRNA 平滑肌细胞 动脉导管 细胞培养 胎兔 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R331.31[医药卫生—基础医学]