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目的通过干扰人脐静脉内皮细胞(HUVEC)S100A4基因表达,探讨其对晚期糖基化终产物(AGE)诱导HUVEC钙化的影响。方法用重组病毒转染体外培养的HUVEC抑制S100A4的表达。用AGE刺激转感染后的HUVEC,建立糖尿病细胞模型。通过检测细胞内钙离子水平,Western blotting测定基质Gla蛋白(MGP)、骨形态发生蛋白2(BMP2)、碱性磷酸酶(ALP)的表达水平,以及茜素红染色评估HUVEC钙化水平。结果在AGE的刺激下HUVEC内钙离子浓度显著升高。沉默S100A4基因后,可以反转AGE诱导的MGP高表达,以及BMP2、ALP的低表达(均P<0.01);且茜素红染色面积明显降低。结论用重组病毒沉默S100A4基因可以降低AGE诱导的HUVEC钙化水平。S100A4基因可能成为治疗糖尿病血管钙化的一个潜在的靶点。
Objective To investigate the effect of endothelin (S100A4) on the expression of S100A4 in human umbilical vein endothelial cells (HUVECs) induced by advanced glycation end products (AGE). Methods The recombinant virus was transfected into HUVECs in vitro to inhibit the expression of S100A4. The transfected HUVECs were stimulated with AGE to establish diabetic cell model. The levels of intracellular Ca2 + were measured by Western blotting, and the expression of matrix glycoprotein (MGP), bone morphogenetic protein 2 (BMP2) and alkaline phosphatase (ALP) were detected by Western blotting. The calcification of HUVECs was evaluated by alizarin red staining. The results of AGE stimulated HUVEC intracellular calcium concentration was significantly increased. Silencing the S100A4 gene could reverse AGE-induced MGP overexpression and BMP2 and ALP expression (all P <0.01), and the area of alizarin red staining was significantly reduced. Conclusion Silencing S100A4 gene with recombinant virus can reduce AGE-induced HUVEC calcification. The S100A4 gene may be a potential target for the treatment of diabetic vascular calcification.