钙调神经磷酸酶对钙化细胞Ⅰ型三磷酸肌醇受体表达的影响
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Regulation of Expression of 1,4,5 Trisphosphate ReceptorⅠby Calcineurin in Cellular Calcification
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    摘要:

    目的探讨大鼠血管平滑肌细胞钙化过程中钙调神经磷酸酶对Ⅰ型三磷酸肌醇受体蛋白和mRNA表达的影响。方法大鼠胸主动脉血管平滑肌细胞经传代培养后随机分为对照组、钙化组和环孢素A组,采用磷酸二氢钠诱导大鼠动脉血管平滑肌细胞钙化。应用免疫印迹法及实时荧光定量RT-PCR法测定钙调神经磷酸酶B亚基、Ⅰ型三磷酸肌醇受体的表达变化,同时测定Ca2+浓度及碱性磷酸酶、钙调神经磷酸酶活性。结果与对照组比较,钙化组钙调神经磷酸酶B亚基、Ⅰ型三磷酸肌醇受体蛋白和mRNA表达显著增加(P<0.01);与钙化组比较,环孢素A组钙调神经磷酸酶B亚基、Ⅰ型三磷酸肌醇受体蛋白和mRNA表达显著降低(P<0.01)。钙化组Ca2+浓度、钙调神经磷酸酶活性、碱性磷酸酶活性较对照组显著增高(P<0.01);环孢素A组碱性磷酸酶活性、Ca2+浓度较钙化组显著增加(P<0.05或P<0.01),钙调神经磷酸酶活性较钙化组显著降低(P<0.01)。结论钙调神经磷酸酶能够增强钙化细胞中Ⅰ型三磷酸肌醇受体mRNA和蛋白的表达。

    Abstract:

    Aim To explore the modification of inositol 1,4,5-trisphosphate receptor type Ⅰ(IP3RⅠ) expression by calcineurin in calcification of vascular smooth muscle cells. Methods Rat aortic vascular smooth muscle cells after subculture were randomly divided into control group,calcification group and cyclosporine A(CsA) group.Rat aortic vascular smooth muscle cell calcification model was constructed by NaH2PO4.Expression of IP3RⅠand calcineurin was determined by RT-PCR and Western blot methods.Alkaline phosphatase(ALP) Activity and Ca2+ concentration were determined by colorimetric method.CaN activity was determined by ELISA. Results Expressions of IP3RⅠand CaNB mRNA and protein increased in calcification group compared with control group(P<0.01).Expressions of IP3RⅠand CaNB mRNA and protein decreased in CsA group compared with calcification group(P<0.01).Ca2+ concentration and CaN and ALP activities of rat aortic smooth muscle cells increased in calcification group compared with control group(P<0.01).ALP activity and Ca2+ concentration increased in CsA group compared with calcification group(P<0.05 or P<0.01).CaN activity decreased in CsA group compared with calcification group(P<0.01). Conclusions CaN can enhance the expression of IP3RⅠmRNA and protein in cell calcification.

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李芳,李绍冰,苑晓烨,姚丽霞,田一帆,冯会英,李秀芹,孙鹏.钙调神经磷酸酶对钙化细胞Ⅰ型三磷酸肌醇受体表达的影响[J].中国动脉硬化杂志,2011,19(12):1001~1004.

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  • 收稿日期:2011-04-01
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