Investigation for the Mechanism of Vascular Smooth Muscle Cell Calcification Induced by Calcium and Phosphorus in Experimental Rats

Objective: To explore the effect of oxidative stress injury on the mechanism of vascular smooth muscle cell(VSMC) calcification induced by calcium and phosphorus in experimental rats. Methods: The VSMC calcification was induced by incubating the cells with calcium chloride(Ca Cl2) and β-sodium glycerophosphate(β-GP) for 8 days, and the cells were divided into 4 groups: 1 Control group, 2 Calcification group, 3 Calcification + H2O2 group, 4 Calcification + catalase group. The calcium nodule formation and calcium deposition in VSMC were detected by Alizarin red staining and o-cresolphthalein complexone method, the reactive oxygen species(ROS) was detected by DCFH-DA probe staining and the protein expression of Runx2 was examined by Western blot analysis. Results: Compared with Control group, Calcification group showed the higher ROS production, more calcium nodule and calcium deposition, higher Runx2 protein expression; while compared with Calcification group, the above indexes were even higher in Calcification + H2O2 group, P0.05. The ROS production, calcium nodule, calcium deposition and Runx2 protein expression were lower in Calcification + catalase group than those in Calcification group and Calcification + H2O2 group, but still higher than that in Control group. The protein expression of Runx2 was similar between Calcification + catalase group and Control group, P0.05.Conclusion: Ca Cl2 and β-GP treatment may induce VSMC calcification via activating ROS-Runx2 signal pathway in experimental rats.