白细胞介素10对体外培养大鼠主动脉平滑肌细胞成骨样分化与钙化的影响

Role of interleukin 10 in osteogenic transition and calcification of rat vascular smooth muscle cells in vitro

目的探究白细胞介素10(IL?10)对体外培养大鼠主动脉平滑肌细胞(VSMCs)成骨分化,钙化和可能的信号传导途径。方法提取大鼠胸VSMCs,采用钙浓度测定,碱性磷酸酶(ALP)活性测定和茜素红染色观察IL?10对钙化的影响。实时聚合酶链反应(Real?Time PCR)探究IL?10对成骨样分化的作用;蛋白质免疫印迹(Western Blot)用于检测成骨分化蛋白,观察IL?10对高钙高磷诱导的VSMCs钙化的作用。结果IL?10促进高钙高磷引起的VSMCs钙化,上调成骨分化标志物的表达,激活骨形态形成蛋白2/白细胞抑制因子1,5/Runt相关转录因子2通路(BMP2/Smad1,5/RUNX2),并且抑制活化T细胞核因子c1(NFATc1)的表达。结论IL?10能够诱导VSMCs成骨样分化,这可能是临床上观察到IL?10与血管钙化相关的机制之一。IL?10这一作用与其激活BMP2/Smad1,5/RUNX2通路,抑制NFATc1激活有关。

Objective To investigate the role of interleukin 10(IL?10)in osteogenic transition and calcification of vascular smooth muscle cells(VSMCs),and the possible cell signal transduction way.Methods Rat VSMCs were isolated.The calcium quantifica?tion,alkaline phosphatase activation and Alizarin red staining were performed to determine the effect of IL?10 on calcification,and the mRNA expression of osteogenic markers were determined by RT?PCR,Western Blot.Results IL?10 promoted VSMCs calcifica?tion induced by high calcium and phosphate,enhanced the mRNA expression of osteogenic markers,activated the BMP2/Smad1,5/RUNX2 pathway and decreased the expression of NFATc1.Conclusion IL?10 could induce osteogenic transition in vitro,which may be one of the mechanism involved in IL?10 associated vascular calcification as observed in clinical studies.The role of IL?10 in VSMCs may partially achieved through activating BMP2/Smad1,5/RUNX2 cell signal transduction wayand decreasing the expres?sion of NFATc1.