糖基化终产物对人血管平滑肌细胞表达及分泌甲状旁腺激素相关肽和血管钙化的影响-东南大学学报医学版

糖基化终产物对人血管平滑肌细胞表达及分泌甲状旁腺激素相关肽和血管钙化的影响

单位：1. 东南大学医学院, 江苏南京 210009;
2. 东南大学附属中大医院心血管内科, 江苏南京 210009

分类号：R587.1;R543.5;R-33

出版年·卷·期（页码）：2014·33·第五期（545-550）

摘要：

目的：观察糖基化终末产物（AGEs）对人血管平滑肌细胞表达及分泌甲状旁腺激素相关肽（PTHrP）功能的影响，探讨PTHrP影响血管平滑肌细胞钙化发生的相关机制。方法：不同浓度的AGE-BSA分别与人血管平滑肌细胞孵育相同时间后，检测各组细胞钙含量及碱性磷酸酶（ALP）活性判断钙化程度；酶联免疫吸附法检测各组细胞PTHrP的分泌量；实时荧光定量逆转录聚合酶链反应检测PTHrP、核心结合因子（cbfα1）及骨形态发生蛋白2（BMP-2）在各组细胞中的表达量。结果：随AGEs浓度增加，细胞中的钙含量及ALP活性增加（P<0.05），而细胞分泌的PTHrP量逐渐减少（P<0.05），且AGEs可促进cbfα1、BMP-2及PTHrP在血管平滑肌细胞的表达，这在较高浓度组较为显著（P<0.05）。结论：适当浓度的晚期AGEs可影响人血管平滑肌细胞PTHrP的表达水平及分泌量，促进钙含量增加，进而有助于血管钙化的发生。

Objective: To investigate the effects of advanced glycation end-products(AGEs) on expression and secretion of parathyroid hormone related peptide(PTHrP) in vitro cultured human vascular smooth muscle cells(HVSMCs), and explore the related mechanism of PTHrP influencing vascular smooth muscle cells calcification. Methods: HVSMCs were treated with AGE-BSA of indicated concentration or non-glycated BSA for same periods. The calcium contents and activity of alkaline phosphatase of cells were analyzed by microplate reader; PTHrP levels in the supernatant were detected by the enzyme-linked immunosorbent method. Real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR) was performed to detect the expressive of PTHrP, core-binding factor α1(cbfα1) in human and bone morphogenetic protein-2(BMP-2) in vascular smooth muscle cells. Results: AGE-BSA increased calcium deposition and activity of alkaline phosphatase in HVSMCs in dose-independent manners(P<0.05), but reduced the secretion of PTHrP. Futhermore, the elevated AGE-BSA treatment on HVSMCs significantly enhanced the expression of cbfα1, BMP-2 and PTHrP, compared with the controls(P<0.05). Conclusion: The expression and secretion of PTHrP on human vascular smooth muscle cells can be effected by AGEs, and PTHrP may induce deposition of calcium on vascular smooth muscle cells, thereby contributing to the vascular calcification. However, the related mechanism will be further explored.

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