哇巴因对自发性高血压大鼠和Wistar-Kyoto大鼠动脉平滑肌细胞钠泵基因表达的影响  

Effects of Ouabain on Na^+-K^+-ATPase Activity and mRNA Expression in Cultured Aortic Smooth Muscle Cells from Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

作　　者：李成浩[1,3] 商黔惠[1] 姜黔峰[1] 刘祖林 万卫红 

机构地区：[1]遵义医学院临床医学研究所,遵义医学院附属医院心血管内科,贵州遵义563003 [2]贵州省细胞工程重点实验室,贵州遵义563003 [3]湖北省襄樊市中医院心血管内科

出　　处：《中华高血压杂志》2010年第6期559-564,共6页Chinese Journal of Hypertension

基　　金：贵州省科学技术基金重点项目;贵州省优秀科技教育人才省长专项资金[(2002)3013,(2005)239]

摘　　要：目的研究钠泵(Na+-K+-ATPase)抑制剂哇巴因对自发性高血压大鼠(SHR)和Wistar-Kyoto(WKY)大鼠动脉平滑肌细胞钠泵活性及其α1、β1亚单位mRNA表达的影响。方法体外培养14周龄SHR和WKY大鼠动脉平滑肌细胞,以无血清培养基培养的动脉平滑肌细胞作为对照,先向培养液中加入终浓度为1×10-7mol/L或1×10-5mol/L的哇巴因,分别孵育1、6、24h后收集细胞,再向培养液中分别加入不同终浓度的哇巴因(1×10-9、1×10-7、1×10-5mol/L)孵育24h后收集细胞。用生化酶学方法测定钠泵活性,用逆转录聚合酶链反应技术测定钠泵α1、β1亚单位mRNA相对表达量。结果 SHR动脉平滑肌细胞钠泵活性与WKY比较显著降低(P<0.05);SHR动脉平滑肌细胞钠泵α1、β1亚单位mRNA表达均低于WKY大鼠(均P<0.05)。与对照组比较,哇巴因1×10-5mol/L干预24h后,SHR钠泵活性降低(P<0.05),α1、β1亚单位mRNA表达下调(均P<0.05),WKY组钠泵活性增高(P<0.05),α1、β1亚单位mRNA表达上调(均P<0.05);其改变没有明显的浓度依赖性。结论 SHR动脉平滑肌细胞钠泵活性下降可能部分与α1、β1亚单位mRNA表达下调有关。哇巴因下调SHR动脉平滑肌细胞钠泵活性及α1、β1亚单位mRNA表达,而上调WKY大鼠动脉平滑肌细胞钠泵活性及α1、β1亚单位mRNA表达。Objective To investigate the effects of ouabain on Na＋-K＋-ATPase activity and the mRNA expression of α1,β1 subunit in cultured aortic smooth muscle cells （ASMCs） from spontaneously hypertensive rats（SHR） and Wistar-Kyoto（WKY） rats. Methods ASMCs from 14-week-old SHR and WKY were investigated as target cells,whereas cultured ASMCs derived from SHR and WKY in serum-free medium were used as control groups. Na＋-K＋-ATPase activity was determined biochemically. One-step reverse transcription-polymerase chain reaction （one step RT-PCR） technique were used to detect the effects of ouabain on the α1,β1 subunits mRNA expression level. Results The activity of Na＋-K＋-ATPase and the mRNA expression of Na＋-K＋-ATPase α1 and β1 subunits were significantly decreased in SHR as compared with WKY group （P〈0.05）. Ouabain distinctly attenuated the activity of Na＋-K＋-ATPase（P〈0.05）,downregulated α1,β1 subunits mRNA expression in SHR ASMCs （P〈0.05）. However,Na＋-K＋-ATPase activity was increased in WKY rats ASMCs（P〈0.05）. Conclusion The decreased activity of Na＋-K＋-ATPase in SHR may be partly related to suppression of the mRNA expression of α1 and β1 subunits. Ouabain inhibits Na＋-K＋-ATPase activity and downregulates α1,β1 subunits mRNA expression from SHR ASMCs,whereas,ouabain increases Na＋-K＋-ATPase activity and upregulates mRNA expression of α1,β1 subunits in ASMCs derived from WKY rats.

关 键 词：哇巴因 钠泵 动脉平滑肌细胞 高血压 

分 类 号：R544.1[医药卫生—心血管疾病]