S100A4基因沉默抑制晚期糖基化终产物诱导内皮细胞钙化的作用

Inhibition of advanced glycation end products-induced endothelial cells calcification by silencing S100A4

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 effects of silencing the S100A4 gene of human umbilical vein endothelial cells （HUVECs） on advanced glycation end products （AGE）-induced vascular calcification. Methods Recombinant viruses were transfected into HUVECs cultured in vitro to inhibit the expression of S100A4. Transfected HUVECs were stimulated with AGE to construct a diabetic cell model. The calcification of HUVECs was assessed by measuring the calcium ion level in ceils, detecting the expression levels of matrix Gla protein （MGP）, bone morphogenetic protein-2 （BMP2）, and alkaline phosphatase （ALP） with Western blotting, as well as alizarin red staining. Results The concentration of calcium ion in HUVECs was significantly increased by the stimulation of AGE. Silencing of S100A4 could reverse the AGE-induced high expression of MGP and AGE-induced low expressions of BMP2 and ALP （P〈0.01 for all） and significandy reduce the area of alizarin red staining. Conclusion Silencing of S100A4 with recombinant viruses can reduce the calcification of HUVEC induced by AGE. SIOOA4 may be a potential target for the treatment of vascular calcification in diabetic Patients.