维生素K_2对大鼠主动脉钙化的作用研究

Effect of Vitamin K_2 on Theaortic Artery Calcification in Experimental Rats

目的:探讨维生素K_2对大鼠血管钙化形成及氧化应激损伤的作用。方法:24只大鼠随机分为4组:对照组、6周钙化组、12周钙化组和6周钙化+6周维生素K_2组。采用华法林诱导大鼠体内血管钙化形成。分别通过茜素红S染包法和邻甲酚肽络合酮比包法检测4组大鼠主动脉组织钙结节形成情况及钙沉积含量;活性氧检测试剂盒(二氢乙啶)检测大鼠主动脉组织活性氧阳性细胞数;透射电子显微镜检测大鼠血管平滑肌细胞线粒体形态变化。结果:6周及12周两个钙化组大鼠主动脉有钙结节形成,钙沉积含量及活性氧水平均显著高于对照组(P均<0.01);6周钙化+6周维生素K_2组大鼠与两个钙化组相比,上述各指标均显著降低,差异均有统计学意义(P均<0.01)。两个钙化组大鼠血管平滑肌细胞线粒体肿胀,结构模糊不清,胞质内空泡变性;6周钙化+6周维生素K_2组大鼠血管平滑肌细胞体积与对照组比较无明显变化,胞质内未见空泡变性。结论:华法林诱导血管钙化形成与氧化应激损伤有关,氧化应激损伤可造成细胞超微结构损伤。维生素K_2可能通过减轻血管平滑肌细胞氧化应激损伤,改善血管钙化。

Objective： To explore the effects of Vitamin K2 （VK2） on theaortic artery calciifcation and oxidative stress injury in experimental rats. Methods： A total of 24 rats were divided into 4 groups：①Control group,②6-week calciifcation group,③12-week calciifcation group and④6-week calciifcation ＋ 6-week VK2 group;n=6 in each group. The arterial calciifcation was induced by warfarin （WFN） treatment. The calcium nodule and deposition in rat’s theaortic artery were detected by Alizarin red staining and o-cresolphthalein complexone method, the reactive oxygen species （ROS） were measured by DHE probe staining and the morphological changes of mitochondria in smooth muscle cells were detected by transmission electron microscopy. Results： Calciifcation nodule formed in both 6-week and 12-week calciifcation groups, the calciifcation deposition and ROS were higher than Control group,P〈0.01. Compared with both calcification groups, the above indexes were decreased in 6-week calciifcation ＋ 6-week VK2 group,P〈0.01. Both calciifcation groups showed mitochondria swelling with unclear structure and cytoplasm vacuoles degeneration in vascular smooth muscle cells. The vascular smooth muscle cell volumes were similar between Control group and 6-week calcification ＋ 6-week VK2 group, and no cytoplasm vacuoles degeneration was observed. Conclusion： Warfarin induced aortic calciifcation is related to oxidative stress injury which may cause the ultra-micro structural damage in smooth muscle cells; VK2 may reduce the oxidative stress injury and improve the condition of vessel calciifcation in experimental rats.