Vascular calcification is closely linked to patients in diabetes mellitus and chronic kidney disease. Advanced glycation end-products (AGEs) are associated with osteogenic differentiation of vascular smooth muscle cell (VSMC), vascular calcification, and autophagy that takes part in the process. However, the underlying mechanism of the effects of AGEs on the phenotypic transition and autophagy of VSMCs is not clearly understood. In this study, we cultured the rat VSMC line (A7R5) and thoracic aorta organ with bovine serum albumin (BSA) or AGEs (AGEs-BSA) and detected proteins expression by Western blotting or immunofluorescence. Autophagosome was observed by transmission electron microscopy (TEM). The mineralization and calcific nodules were identified by Alizarin Red S and Von Kossa staining. AGEs significantly downregulated p-AMPKα expression and upregulated p-mTOR expression and then increased the expression of osteoblastic differentiation, while suppressing autophagy in a time-dependent pattern. Pretreatment with autophagy activator rapamycin and AMPK activator AICAR both upregulated the autophagy level and downregulated the effects of AGEs on osteoblastic differentiation of VSMCs. Moreover, the result from rat thoracic aorta culture also confirmed that AGEs promote vascular calcification in a time-dependent manner. Thus, our study showed that AGEs quicken vascular calcification and suppress autophagy associated with AMPK/mTOR signaling pathway.

血管钙化与糖尿病和慢性肾脏病患者密切相关。晚期糖基化终产物（AGEs）与参与该过程的血管平滑肌细胞（VSMC）的成骨分化，血管钙化和自噬有关。然而，尚不清楚AGEs对VSMCs的表型转变和自噬的潜在机制。在这项研究中，我们用牛血清白蛋白（BSA）或AGEs（AGEs-BSA）培养了大鼠VSMC系（A7R5）和胸主动脉器官，并通过Western印迹或免疫荧光检测了蛋白表达。通过透射电子显微镜（TEM）观察自噬体。通过茜素红S和Von Kossa染色鉴定了矿化和钙化结节。AGEs显着下调p-AMPKα表达并上调p-mTOR表达，然后增加成骨细胞分化的表达，同时以时间依赖性方式抑制自噬。自噬激活剂雷帕霉素和AMPK激活剂AICAR的预处理均上调了自噬水平，并下调了AGEs对VSMC成骨细胞分化的影响。此外，大鼠胸主动脉培养的结果也证实AGEs以时间依赖性方式促进血管钙化。因此，我们的研究表明，AGEs可以加速血管钙化并抑制与AMPK / mTOR信号通路相关的自噬。自噬激活剂雷帕霉素和AMPK激活剂AICAR的预处理均上调了自噬水平，并下调了AGEs对VSMC成骨细胞分化的影响。此外，大鼠胸主动脉培养的结果也证实AGEs以时间依赖性方式促进血管钙化。因此，我们的研究表明，AGEs可以加速血管钙化并抑制与AMPK / mTOR信号通路相关的自噬。用自噬活化剂雷帕霉素和AMPK活化剂AICAR预处理均上调了自噬水平，并下调了AGEs对VSMC成骨细胞分化的影响。此外，大鼠胸主动脉培养的结果也证实AGEs以时间依赖性方式促进血管钙化。因此，我们的研究表明，AGEs可以加速血管钙化并抑制与AMPK / mTOR信号通路相关的自噬。