甲基奈醌-4对血管平滑肌细胞钙化的影响

Effect of Menaquinone-4 on vascular smooth muscle cell calcification

目的探讨甲基奈醌-4对血管平滑肌细胞钙化的影响。方法分别采用生长培养基(磷1. 4 mmol/L)和钙化培养基(磷3. 0 mmol/L),培养人脐静脉血管平滑肌细胞,使用甲基酚酞络合铜法检测细胞基质钙、比色法检测碱性磷酸酶活性、液相色谱法检测细胞内甲基奈醌-4水平,茜素红S染色观察细胞基质钙盐沉积。在钙化培养基下,改变甲基奈醌-4水平,观察细胞基质钙、碱性磷酸酶活性的变化。结果钙化培养基能增加人脐静脉血管平滑肌细胞基质钙含量、碱性磷酸酶活性,减少细胞内甲基奈醌-4水平,细胞外基质出现明显的钙盐沉积。减少甲基奈醌-4的水平能增加基质钙含量、碱性磷酸酶活性。添加甲基奈醌-4能减少基质钙含量、碱性磷酸酶活性。结论甲基奈醌-4能抑制磷诱导的血管平滑肌细胞钙化。

Objective To investigate the effect of menaquinone-4 on vascular smooth muscle cell calcification. Methods With either growth medium( 1. 4 mmol/L,Pi) or calcification medium( 3. 0 mmol/L,Pi),primary human umbilical vein smooth muscle cells( HUVSMCs)were incubated. Cell matrix calcium was quantitated using o-cresolphthalein complexone method. Alkaline phosphatase activity was assessed colorimetrically. Menaquinone-4 was measured by ultra-performance liquid chromatography-tandem mass spectrometry. Matrix calcification morphology was by Alizarin Red S staining. Matrix calcium morphology and alkaline phosphatase activity were observed under different levels of Menaquinone-4. Results Calcification medium increased matrix calcium,alkaline phosphatase activity and reduced menaquinone-4 level compared with growth medium( P < 0. 05). Abundant mineral deposits could be detected in the extracellular matrix upon calcification medium. Decreased menaquinone-4 in HUVSMCs increased matrix calcification and alkaline phosphatase activity,and vice-versa. Conclusion Menaquinone-4 might inhibit phosphate-induced vascular smooth cells calcification.