Advanced glycation endproducts regulate smooth muscle cells calcification in cultured HSMCs.
Huqiang He, Yong Liu, Hong Zeng, Xiaolei Sun, Lei Zhang, Xue-Lin Zhang, Wenjun Liao, Xiangyu Zhou, Yanzheng He
Abstract
Huqiang He, Yong Liu, Hong Zeng, Xiaolei Sun, Lei Zhang, Xue-Lin Zhang, Wenjun Liao, Xiangyu Zhou, Yanzheng He
Abstract
OBJECTIVE: To investigate the mechanism of Advanced glycation end products (AGEs) promoting the calcification of smooth muscle cells. METHODS: The successfully cultured smooth muscle cells were divided into three groups: normal culture group (group A), calcified culture group (group B), calcification + AGEs group (group C); the concentration of intracellular calcium ion was detected in each group; the promotion of AGEs on the calcification of HSMCs was confirmed by VON KOSSA staining; and the expressions of β-catenin, RAGE, β-catenin, OPG and E-cadherin protein were detected by immunofluorescence and western blot. RESULTS: The morphology of the cells in each group showed that the amount of calcified plaques in calcification + AGES group were significantly higher than the calcification group. VON KOSSA staining showed that with increasing concentrations of AGE-BSA, the amount of its calcification gradually increased. Calcium concentration in Calcification + 20 mg/L AGEs group was significantly higher, followed by 40 mg/L AGEs group. The expression of β-catenin increased with the increasing concentrations of AGEs. CONCLUSION: AGEs can promote the calcification of human femoral artery smooth muscle cells, with a concentration gradient effect. With increasing concentrations of AGEs, the expression of RAGE increased, indicating that AGEs-induced HSMCs proliferation was correlated with RAGE expression.
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OBJECTIVE: To investigate the mechanism of Advanced glycation end products (AGEs) promoting the calcification of smooth muscle cells. METHODS: The successfully cultured smooth muscle cells were divided into three groups: normal culture group (group A), calcified culture group (group B), calcification + AGEs group (group C); the concentration of intracellular calcium ion was detected in each group; the promotion of AGEs on the calcification of HSMCs was confirmed by VON KOSSA staining; and the expressions of β-catenin, RAGE, β-catenin, OPG and E-cadherin protein were detected by immunofluorescence and western blot. RESULTS: The morphology of the cells in each group showed that the amount of calcified plaques in calcification + AGES group were significantly higher than the calcification group. VON KOSSA staining showed that with increasing concentrations of AGE-BSA, the amount of its calcification gradually increased. Calcium concentration in Calcification + 20 mg/L AGEs group was significantly higher, followed by 40 mg/L AGEs group. The expression of β-catenin increased with the increasing concentrations of AGEs. CONCLUSION: AGEs can promote the calcification of human femoral artery smooth muscle cells, with a concentration gradient effect. With increasing concentrations of AGEs, the expression of RAGE increased, indicating that AGEs-induced HSMCs proliferation was correlated with RAGE expression.
Key concepts: Von Kossa stain, Calcification, Glycation, Staining, Western blot, Chemistry, Internal medicine, Endocrinology