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不同浓度β-甘油磷酸盐对人脐静脉平滑肌细胞钙化的影响

Keywords: 人脐静脉血管平滑肌细胞,β-甘油磷酸盐,血管钙化

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Abstract:

目的探寻β-甘油磷酸盐诱导原代培养人脐静脉平滑肌细胞(HUSMC)钙化的适宜浓度。方法采用原代培养人脐静脉平滑肌细胞,根据培养液中β-甘油磷酸盐浓度不同依次分为8组正常对照组、10mM组、12mM组、14mM组、16mM组。以更换培养液当天记为第1天,培养10d,vonkossa染色观察细胞钙沉积情况、显微图像分析法测量钙化阳性细胞百分比、比色法检测细胞内钙含量和碱性磷酸酶活性。结果对照组SMC银染无钙结节,显微图像分析阳性细胞为(0.1060±0.05)%;12mM组SMC银染可见明显钙结节存在,钙化阳性细胞数为(4.7009±1.35)%,钙含量和碱性磷酸酶活性较对照组明显升高(P0.05),但10mM组、16mM组钙化阳性细胞百分比升高明显(P0.05);40mM组钙含量和碱性磷酸酶活性明显降低(P<0.001),灰度扫描钙化阳性细胞随之降低。结论12mMβ-甘油磷酸盐可刺激人脐静脉平滑肌细胞出现钙盐沉积,且人脐静脉SMC钙化对β-甘油磷酸盐无浓度依赖性。

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