目的探讨丹参多酚酸盐对大鼠血管平滑肌细胞(vascularsmoothmusclecells，VSMCs)钙化的抑制作用。方法实验分钙化组(β-磷酸甘油诱导培养大鼠VSMCs钙化)，钙化+不同浓度的丹参多酚酸盐组和正常对照组(普通培养液)。通过细胞钙含量和碱性磷酸酶(ALP)活性测定，判断钙化程度;分别用real-timePCR方法和westernblot方法检测细胞中骨形成蛋白BMP-2的mRNA水平和蛋白水平。结果钙化VSMCs的钙含量和ALP活性分别增加3.92倍和10.5倍(均P<0.01)。而丹参多酚酸盐呈剂量依赖的抑制钙化的VSMCs中的钙含量升高和ALP活性，其中10-8、10-7、10-6M的丹参多酚酸盐使钙化量分别减低32.3%、42.2%和60.9%(P<0.01)，使ALP活性分别减低36.4%、48.8%和74.7%(P<0.01)。钙化的VSMCs中BMP-2mRNA和蛋白水平较对照组均有显著增高，而丹参多酚酸盐呈剂量依赖的抑制钙化的VSMCs中BMP-2mRNA表达和蛋白水平。结论丹参多酚酸盐可以抑制大鼠的VSMCs发生钙化，此效应可能与降低BMP-2表达有关。
Abstract
Objective To study the effects of salvianolate on the calcification in rat vascular smooth muscle cells (VSMCs). Methods Calcification of cultured rat VSMCs was prepared by incubation with β-glycerophosphate. Calcification was confirmed by measurement of calcium content and alkaline phosphatase (ALP) activity in VSMCs. The mRNA and protein levels of BMP-2 were detected by using real-time PCR and Western blotting, respectively. Results Compared with control group, 3. 92- fold calcium content and 10.5-fold increase in ALP activity were observed in calcified VSMCs group, respectively. Levels of mRNA and protein of BMP-2 were significantly increased in calcified group. Salvianolate decreased the calcium and ALP activity in a dose-dependent manner, these data were decreased by 32.3 %, 42.2%, 60.9%(P<0.05); 36.4%, 48.8% and 74.7% (P<0.01), respectively. And salvianolate decreased the mRNA and protein levels of BMP-2. Conclusion Salvianolate may have cardiovascular protecting effect by antagonizing vascular calcification, maybe by regulating BMP-2 level.
关键词
丹参多酚酸盐 /
钙化 /
血管平滑肌细胞 /
骨形成蛋白
Key words
salvianolate /
calcification /
vascular smooth muscle cells /
BMP-2
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参考文献
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