论文摘要
目的:探讨E-选择素（E-selectin）和细胞间粘附分子-1（ICAM-1）反义寡核苷酸（asODN）及其与辛伐他汀（simvastatin）联合应用对人脐静脉内皮细胞（HUVEC） ICAM-1和E-selectin表达的抑制和对外周血单个核细胞粘附的影响。方法:原代培养人脐静脉内皮细胞（HUVEC）,并传代;应用脂质体转染法将人工合成E-selectin asODN和ICAM-1 asODN分别导入HUVECs;实验共分8组,分别为空白对照（basal）组、TNF-α诱导组、liposome对照组、liposome-control asODN组、裸asODN （nude asODN）组、脂质体asODN （lipo-asODN）组、simvastatin处理组、lipo-asODN与simvastatin联合组,除空白组外,其余7组均以TNF-α诱导内皮损伤;以流式细胞技术测定HUVECs细胞膜E-selectin和ICAM-1蛋白的表达;以RT-PCR半定量技术测定HUVECs E-selectin mRNA和ICAM-1 mRNA的表达;以细胞粘附试验测定HUVECs与单个核细胞的粘附率;应用辛伐他汀预先处理HUVECs,再用E-selectin lipo-asODN和ICAM-1 lipo-asODN转染HUVECs,观察E-selectin asODN和ICAM-1 asODN与辛伐他汀联合应用对HUVECs E-selectin和ICAM-1的表达以及对HUVECs与外周血单个核细胞粘附抑制的影响。结果:⑴与TNF-α诱导组相比,E-selectin asODN和ICAM-1 asODN以及E-selectin lipo-asODN和ICAM-1 lipo-asODN均明显降低了HUVECs相应的E-selectin和ICAM-1蛋白及mRNA的表达（P<0.01）;⑵E-selectin asODN和ICAM-1 lipo-asODN与simvastatin联合应用可降低对HUVECs相应的E-selectin和ICAM-1的蛋白及mRNA表达;⑶lipo-asODN与simvastatin联合组与1ipo-asODN组比较,E-selectin表达水平显著下降（P<0.01）,而ICAM-1的表达水平下降无显著意义（P>0.05）,表明E-selectin lipo-asODN与simvastatin联合作用有更好的抑制效果。⑷与TNF-α刺激组相比,lipo-asODN组及1ipo-asODN与simvastatin联合组HUVECs与单个核细胞粘附率均显著下降（P<0.01）,尤其以HUVECs与CD3+T淋巴细胞的粘附率下降最为明显,但1ipo-asODN与simvastatin联合组与1ipo-asODN组比较,未见明显变化（P>0.05）。结论:E-selectin asODN和ICAM-1 asODN可抑制HUVECs相应的蛋白及mRNA的表达,并可抑制HUVECs与单个核细胞的粘附,尤其对HUVECs与CD3+ T细胞粘附的抑制作用更为明显。1ipo-asODN与simvastatin联合应用比单独1ipo-asODN的抑制作用更加明显。本研究为E-selectin asODN和ICAM-1 asODN应用于临床抗动脉粥样硬化的基因治疗的前景开拓新的思路和提供理论依据。
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标签：选择素论文; 细胞间粘附分子论文; 反义寡核苷酸论文; 辛伐他汀论文; 人脐静脉内皮细胞论文;