CD73对动脉粥样硬化中血管平滑肌细胞的作用

 目的  探讨CD73对血管平滑肌细胞单核细胞趋化蛋白-1(moncyte chemoattractant protein-1,MCP-1)、IL-8和基质金属蛋白酶-1(matrix metalloproteinase-1,MMP-1)表达及迁移能力的影响,并分析其与动脉粥样硬化发生发展的关系。方法  原代培养人脐带动脉平滑肌细胞 (human umbilical artery smooth muscle cells,HUASMC),利用RNA干扰技术抑制细胞中CD73的表达,以未转染组和阴性转染组为对照。实时荧光定量PCR和ELISA方法检测细胞表达及分泌MCP-1和IL-8 的水平。实时荧光定量PCR和Western blot方法检测细胞表达MMP-1的情况。利用划痕实验观察CD73干扰对细胞迁移能力的影响。结果  成功培养原代HUASMC,并且CD73 siRNA转染可显著降低细胞内CD73表达水平。干扰HUASMC的CD73表达后,细胞表达和分泌MCP-1及IL-8的能力升高,细胞内MMP-1的表达升高,而细胞的迁移能力下降。结论  干扰CD73表达可以使HUASMC的MCP-1、IL-8及MMP-1表达升高;干扰CD73表达可降低HUASMC的迁移,提示CD73在动脉粥样硬化的发生发展过程中发挥重要的生物学作用。

The role of CD73 on vascular smooth muscle cells in atherosclerosis

Objective  To investigate the effect of CD73 on the expression of monocyte chemoattractant protein-1 (MCP-1),IL-8 and matrix metalloproteinase-1 (MMP-1) in vascular smooth muscle cells and their migration capacity,and to determine its relationship with atherosclerosis.Methods  Human umbilical artery smooth muscle cells (HUASMC) were cultured using explant attached method.The expression of CD73 in HUASMC was knocked down by siRNA.Blank and negative transfected cells were served as control groups.The mRNA levels and concentrations of MCP-1 and IL-8 in supernatant of siRNA-treated HUASMC were analyzed by real-time PCR and ELISA analysis.The expression of MMP-1 was detected by real-time PCR and Western blot.Migration capacity was assessed by scratch-wound assay.Results  Primary HUASMC were cultured successfully.CD73 siRNA could knock down the expression of CD73 in HUASMC.CD73 siRNA-treated HUASMC showed enhanced MCP-1,IL-8 and MMP-1 levels,and decreased migration capacity.Conclusions  The down regulation of CD73 enhances the expressions of MCP-1,IL-8 and MMP-1 in HUASMC and reduces the migration capability of HUASMC,which indicates that CD73 may play an important role in the development of atherosclerosis.