高血压大鼠动脉平滑肌细胞钙超载与腺苷三磷酸酶被引量：21

Calcium Overload and Adenosinetriphosphatase in Artery Smooth Muscle Cells in Spontaneously Hypertensive Rats

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摘要目的探讨自发性高血压大鼠(SHR)动脉血管平滑肌细胞钙超载与腺苷三磷酸酶(AT-Pase)的关系。方法组织块种植法培养SHR和Wistar-Kyoto(WKY)大鼠胸主动脉平滑肌细胞,应用流式细胞术、生化酶学方法和逆转录聚合酶链反应(RT-PCR)技术等检测SHR和WKY大鼠动脉平滑肌细胞内游离钙浓度([Ca2+]i)、细胞膜Ca2+-ATPase和Na+-K+-ATPase活性及其mRNA表达水平。结果SHR动脉平滑肌细胞[Ca2+]i浓度高于WKY大鼠[(307.7±32.1vs118.9±21.4)nmol/L,P<0.01)],Ca2+-ATPase和Na+-K+-ATPase活性低于WKY大鼠(1.3±0.2vs2.6±0.3;3.1±0.5vs4.9±0.5,P均<0.01),细胞质膜Ca2+-ATPase(plasmamembraneCa2+-ATPase,PMCA)亚型1和Na+-K+-ATPaseα1亚单位mRNA表达低于WKY大鼠(0.231±0.007vs0.403±0.021;0.253±0.028vs0.634±0.033,P均<0.01)。结论SHR动脉平滑肌细胞出现钙超载,其机制可能部分与细胞膜PMCA和Na+-K+-ATPase活性降低有关,两种ATPase功能降低可能是其基因转录水平下调的结果。 Objective To investigate the relationship of calcium overload and adenosine triphosphatase （ ATPase ） in artery smooth muscle cells （ ASMCs） isolated from spontaneously hypertensive rats （SHR）. Methods The thoracic ASMCs isolated from SHR and Wistar-Kyoto （ WKY ） rats were cultured. Intracellular free Ca^2＋ concentration （ [Ca^2＋ ]i ） was detected by flow cytometry. The activity of Ca^2＋ -ATPase and Na^＋-K^＋-ATPase was measured by biochemistry and enzymology. Reverse transcriptase-polymerase chain reaction （RT-PCR） assay was employed to determine the levels of plasma membrane Ca^2＋-ATPase isoform 1 （ PMCA1 ） and Na^＋-K^＋-ATPasea1-subunit mRNA. Results [Ca2＋ -]i in ASMCs of SHR was higher than those from WKY （307.7±32. 1 vs 118.9±21.4 nmol/L, P 〈0.01）. The activities of Ca^2＋-ATPase and Na^＋-K^＋-ATPase in ASMCs from SHR were lower than those from WKY （1.3±0.2 vs 2. 6±0.3; 3. 1±0.5 vs 4.9±0. 5,P〈0.01, respectively）. Meanwhile, the levels of PMCA1 and Na^＋-K^＋-ATPase a1-subunit mRNA were significantly decreased in SHR ASMCs compared with WKY （PMCA1 mRNA 0. 231±0. 007 vs 0. 403±0. 021;Na^＋ -K^＋ ATPase a1-subunit mRNA 0. 253±0. 028 vs 0. 634±0. 033, P〈0.01）. Conclusion Calcium overload in ASMCs of SHR may be partially ascribed to the decreased activities of PMCA and Na^＋-K^＋-ATPase.

作者商黔惠刘晓鹏方宁吴芹姜黔峰杨征

机构地区遵义医学院附属医院心血管内科河南省洛阳市中心医院急救中心遵义医学院附属医院细胞工程实验室遵义医学院药理学教研室

出处《高血压杂志》 CAS CSCD 北大核心 2006年第5期379-384,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自然科学基金项目[编号:(2002)3013 (2001)3091]

关键词高血压肌平滑血管腺苷三磷酸酶基因表达 Hypertension Muscle, smooth, vascular Adenosinetriphosphatase Gene expression

分类号 R544.1 [医药卫生—心血管疾病]

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