Effect of magnesium on calcification of human vascular smooth muscle cells
SUN Wenxue1, ZHANG
1.Departmant of Nephrology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325015; 2.Departmant of Nephrology, the Central Hospital, Shengli Oil Field, Dongying, 257000
Abstract：Objective: To investigate the effect of magnesium on calcification of vascular smooth muscle cells (VSMCs) induced by elevated calcium (Ca++) and phosphate (P) culture. Methods: The fetus’s umbilical artery VSMCs were extracted under aseptic conditions and cultured in vitro. Cells were divided into three groups: normal control group (normal concentration of Ca++ and P, group A), positive control group (high calcium and phosphate, group B), treatment groups (high calcium and phosphate + magnesium sulfate, group C). The cells were cultured for 3 days, then Part (B1, C1) were continued to develop in high calcium and phosphate culture medium, Part (B2, C2) were changed to develop in ordinary solution. Respectively the calcium content of cell layer were determinated in calcium content test in the 4, 7, 10 days. Meanwhile, the protein expression of Cbfa1 and Prps2 were observed by Western bloting. Results: The primary cells were identified, purity of cultures was assessed by positive immunostaining for α-SMA. Compared with negative control group, the sedimentary on the cells layer of experimental group (B) was observed. The calcium content in calcium content test showed that the high calcium and high phosphorus group (B, B1 and B2) increased significantly (P<0.05), compared with the B, B1 and B2 group, magnesium sulfate group (C, C1 and C2) showed a lower intracellular calcium levels. Western blot test results confirmed that: compared with control group (A), the expression of Cbfa1 increased in B group in 24 hours, 72 hours; compared with B group, the expression of Cbfa1 declined in C group in 12 hours, 24 hours, 72 hours. compared with B1, B2 group, the expression of Prps2 increased in C1, C2 group in 4 days, 7 days, 10 days, was statistically significant (P<0.05). Conclusion: High calcium and high phosphorus promote the VSMCs’ calcification. While magnesium has a significant effect to inhibit the VSMCs calcification, activate the Prps2 protein expression, promote the calcification fade.
