RNA干扰内皮细胞分泌白细胞介素6影响平滑肌细胞的迁移

背景：血管支架置入后靶血管部位易发生炎症反应。 目的：利用siRNA技术抑制内皮细胞白细胞介素6的生成,观察其对平滑肌细胞迁移的影响。 方法：采用RT-PCR测定脂多糖刺激EA.HY926细胞表达白细胞介素6 mRNA的时间梯度与浓度梯度,针对白细胞介素6构建短发卡状siRNA真核表达载体pGensil-1.1-白细胞介素6,通过lipofectamine 2000转染EA.HY926,抑制其白细胞介素6的产生。 结果与结论：pGensil-1.1-白细胞介素6转染EA.HY926细胞后,脂多糖刺激下EA.HY926细胞表达的白细胞介素6 mRNA及蛋白明显减少。共培养模型中,转染pGensil-1.1-白细胞介素6的EA.HY926细胞作用下,人脐静脉平滑肌细胞表达的基质金属蛋白酶9 mRNA及蛋白明显降低,结晶紫染色显示人脐静脉平滑肌细胞迁移数量减少。说明siRNA技术可抑制内皮细胞白细胞介素6的生成,并通过降低平滑肌细胞基质金属蛋白酶9的表达减弱平滑肌细胞的迁移能力。

Effects of RNA interference on interleukin-6 expression in endothelial cells and migration of vascular smooth muscle cells

BACKGROUND:Inflammatory reaction easily occurs in target vascular region after implantation of intravascular stent. OBJECTIVE:To investigate the effects of siRNA technique on migration of vascular smooth muscle cells by inhibiting the expression of interleukin-6 (IL-6) in the endothelial cells. METHODS:The time gradient and concentration gradient of the IL-6 mRNA expression in the EA.HY926 cells stimulated by lipopolysaccharide (LPS) were detected by RT-PCR. Short hairpin RNA eukaryotic expression vector targeting IL-6 gene, named pGensil-1.1-IL-6, was constructed and transfected into EA.HY926 cells for intervention of IL-6 expression by lipofectamine 2000. RESULTS AND CONCLUSION:After pGensil-1.1-IL-6 was transfected into EA.HY926 cells, IL-6 mRNA and protein expression in the EA.HY926 cells stimulated by LPS was obviously decreased. In co-culture model, under the effect of EA.HY926 cells transfected by pGensil-1.1-IL-6, matrix metalloproteinase 9 mRNA and protein expression in the human cord venous smooth muscle cells was obviously decreased. The number of human cord venous smooth muscle cells was decreased. These findings suggest that siRNA technique can inhibit the expression of IL-6 in endothelial cells and attenuate the migration of smooth muscle cells by decreasing matrix metalloproteinase 9 expression.