血管紧张素Ⅱ对正常和高血压大鼠主动脉平滑肌细胞腺苷三磷酸酶的影响  

Effects of AngiotensinⅡ on Adenosinetriphosphatases in Aortic Vascular Smooth Muscle Cells from Wistar-Kyoto Rats and Spontaneously Hypertensive Rats

作　　者：张贵海[1] 商黔惠[1] 姜黔峰[1] 万卫红 

机构地区：[1]遵义医学院临床医学研究所附属医院心内科 [2]贵州省细胞工程重点实验室,贵州省遵义市563003

出　　处：《中国动脉硬化杂志》2010年第4期260-264,共5页Chinese Journal of Arteriosclerosis

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：目的探讨血管紧张素Ⅱ对正常血压Wistar-Kyoto大鼠和自发性高血压大鼠胸主动脉平滑肌细胞膜腺苷三磷酸酶(Ca2+-ATPase和Na+,K+-ATPase)活性及基因表达的影响。方法组织块种植法培养14周龄Wistar-Kyoto大鼠和自发性高血压大鼠胸主动脉平滑肌细胞,分别加入含有1×10-9、1×10-8和1×10-7mol/L血管紧张素Ⅱ培养液,共同孵育6h、12h和24h。采用生化酶学方法和逆转录聚合酶链反应技术,检测主动脉平滑肌细胞膜Ca2+-ATPase、Na+,K+-ATPase活性及其mRNA表达水平。结果低、中浓度血管紧张素Ⅱ(1×10-9和1×10-8mol/L)增加Wistar-Kyoto大鼠Ca2+-ATPase活性(P<0.05～P<0.01),与干预时间呈正相关(r=0.340,0.725),24h达最大值,且上调质膜Ca2+-ATPase亚型1mRNA表达(P<0.05～P<0.01);高浓度(1×10-7mol/L)血管紧张素Ⅱ抑制Ca2+-ATPas活性(P<0.05),与干预时间呈负相关(r=-0.348),其24h效应最强,并下调质膜Ca2+-ATPase亚型1mRNA表达(P<0.05)。3种浓度血管紧张素Ⅱ均抑制自发性高血压大鼠Ca2+-ATPase活性(P<0.05～P<0.01),与干预时间呈负相关(r=-0.346,-0.493,-0.759),24h抑制最强,并下调质膜Ca2+-ATPase亚型1mRNA表达(P<0.05～P<0.01)。3种浓度(1×10-9、1×10-8和1×10-7mol/L)血管紧张素Ⅱ依次在24h、12h、6h显著增加Wistar-Kyoto大鼠Na+,K+-ATPase活性(P<0.05～P<0.01),且剂量依赖性增加24hNa+,K+-ATPase活性及上调α1亚单位mRNA表达(P<0.05～P<0.01),与干预时间呈正相关(r=0.425,0.645,0.767)。低、中浓度血管紧张素Ⅱ对自发性高血压大鼠Na+,K+-ATPase活性及α1亚单位mRNA表达均无影响(均P>0.05);高浓度血管紧张素Ⅱ则抑制Na+,K+-ATPase活性(P<0.01),与干预时间呈负相关(r=-0.589),24h达最大效应,且下调α1亚单位mRNA表达(P<0.05)。结论血管紧张素Ⅱ对正常血压大鼠主动脉平滑肌细胞膜Ca2+-ATPase活性及质膜Ca2+-ATPase亚型1mRNA表达有双向作用,并呈剂量依赖性激活Na+,K+-ATPase活性及α1亚单位mRNA表达;抑制高血压大鼠�Aim To explore the effects of angiotensinⅡon the activities of Ca^2＋-ATPase,Na^＋,K^＋-ATPase and mRNA expression levels of the plasma membrane Ca2＋-ATPase isoform 1 （PMCA1） and Na^＋,K＋-ATPase α1-subunit in cultured thoracic aortic vascular smooth muscle cells （ASMC） from Wistar-Kyoto rats and spontaneously hypertensive rats （SHR）. Methods ASMC isolated from 14-week-old male Wistar-Kyoto rats and SHR were cultured,and treated with different concentrations （1×10^-9,1×10^-8,1×10^-7 mol/L） of AngiotensinⅡ. The activities of Ca^2＋-ATPase,Na^＋,K^＋-ATPase were measured by biochemistry and enzymology. RT-PCR assay was employed to determine the relative levels of PMCA1 and Na＋,K＋-ATPase α1-subunit mRNA in ASMC. Results Low and moderate concentration （1×10^-9,1×10^-8 mol/L） of angiotensinⅡsignificantly increased the activity of Ca^2＋-ATPase in ASMC from Wistar-Kyoto rats（P0.05-P0.01）,and the activity of Ca^2＋-ATPase was positively correlated with the intervention time of angiotensinⅡ（ r=0.340,0.725）,reached the maximum at 24 hours,up-regulated PMCA1 mRNA level at the same time （P〈0.05-P〈0.01）; while high concentration （1×10^-7 mol / L） angiotensinⅡ inhibited Ca^2＋-ATPase activity （P0.05）,and the activity of Ca^2 ＋-ATPase was negatively correlated with the intervention time（r=-0.348）,reached the greatest effect at 24 hours,and simultaneously down-regulated PMCA1 mRNA level （P〈0.05）. Angiotens inⅡ（1×10^-9,1×10^-8,1×10^-7mol/L） decreased significantly the activity of Ca^2 ＋-ATPase in ASMC from SHR （P〈0.05 ～P〈0.01）,and the Ca^2 ＋-ATPase activity was negatively correlated with the intervention time （r=-0.346,-0.493,-0.759）,had the strongest effect at 24 hours,and simultaneously attenuated PMCA1 mRNA level. Three concentration of angiotensinⅡ （1×10^-9,1×10^-8,1×10^-7mol/L） stimulated the activity of Na^＋,K^＋-ATPase in ASMC from Wistar-Kyoto rats in turn at24 hours,12 hours,6 hours （P〈0.

关 键 词：大鼠 平滑肌细胞 腺苷三磷酸酶 血管紧张素Ⅱ 高血压 

分 类 号：R363[医药卫生—病理学]