核因子κB的反义和诱骗性寡核苷酸对大鼠球囊损伤后血管狭窄和新生内膜形成的影响
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上海市教委重点学科基金(8990207)资助


Role of the Antisense and Decoy Oligonucleotide of Nuclear Factor-κB in the Vessel Stenosis and Neointima Formation in Balloon-injured Rat Artery
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    摘要:

    探讨核因子κBp6 5亚基的反义和诱骗性寡核苷酸单独或联合作用对大鼠颈动脉球囊损伤细胞间粘附分子、单核细胞趋化因子的作用。SD大鼠随机分为7组,每组分为6个时相点(6h和1、3、5、7、1 4d) ,每个时相点3只大鼠。制作血管球囊损伤模型。相应时间点处死动物。模型组、正义组、诱骗对照组血管内膜面积、中膜面积、内膜中膜在第5天增加,1 4d达到高峰。管腔面积随时相点减小。反义组、诱骗组、反义组+诱骗组干预后,血管上述病理指标明显改善(P<0 .0 5 ) ,反义组+诱骗组较反义组、诱骗组治疗效果更明显(P<0 .0 5 )。内皮细胞间粘附分子、单核细胞趋化因子mRNA表达在血管损伤后6h即可检测到,3、5、7d后持续表达增加,1 4d后表达降低。免疫组织化学检测显示,内皮细胞间粘附分子1、单核细胞趋化因子1蛋白质表达在6个时相点均为阳性染色,1 4d达到高峰;反义组、诱骗组、反义组+诱骗组治疗后,与模型组、正义组、诱骗对照组相比,内皮细胞间粘附分子、单核细胞趋化因子mRNA表达和蛋白合成在各时相点均降低(P<0 .0 5 )。免疫印迹法检测核蛋白抽提物显示,核因子κBp6 5在血管损伤后6h有一定蛋白表达,1d后蛋白表达明显增加,至7d达高峰,1 4d后蛋白表达略降低。反义组、诱骗组、反义组+诱骗组干预后,各时相点蛋白质表达均较模型组、正义组、诱骗对照组减弱(P<0 .0 5 )。核因子κB调控内皮细胞间粘附分子、单核细胞趋化因子基因表达和蛋白质合成;球囊损伤后血管壁细胞增殖在不同时相点有动态变化;脂质体介导局部转染反义、诱骗性寡核苷酸可抑制核因子κB激活对下游基因的调控,两者联合作用效果更为显著

    Abstract:

    Aim To examine the effect of the antisense or/and decoy oligonucleotide of nuclear factor κB (NF κB) on balloon injured intercellular adhesive molecular 1 (ICAM 1) and monocytes chemotactic protein 1 (MCP 1) in the carotid artery of rats in vivo. Methods Sprague Dawley rats underwent balloon dilation injury of the left carotid artery. Rats was divided into 7 groups (n=18) and each group included 6 time points (6 hours and 1,3,5,7,14 days) (n=3). Uninjured artery of the same rat was used to be control. Results In model group, sense group and scramble group, vessel intima area, media area and intima/media ratio increased after 5 days and reached the maximum after 14 days. Whereas lumen area decreased with different time points. Antisense group, decoy group, antisense plus decoy group improved these observational index (p<0.05). The effect of antisense plus decoy group was more obvious than that of antisense group and decoy group alone. ICAM 1 and MCP 1 mRNA expression were examined after 6 hours of artery injury, but not evident after 1 days. They increased expression continuously after 3,5 and 7 days and decreased after 14 days. Comparing with model group, sense group and scramble group, antisense group, decoy group and antisense plus decoy group all lowered ICAM 1 and MCP 1 mRNA expression in every time point (p<0.05). Immunohistochemistry studies revealed ICAM 1 and MCP 1 protein were positive stain within six time points and maximal after 14 days. In antisense group, decoy group and antisense plus decoy group, ICAM 1 and MCP 1 protein synthesis decreased in every time point comparing with model group, sense group and scramble group. Western blot studies showed NF κB p65 was disperse positive stain after 6 hours of injury and increased after 1 day and reached the peak, but protein expression was weak after 14 days. Antisense group, decoy group and antisense plus decoy group treatment inhibited protein synthesis more significantly than those of model group, sense group and scramble group(p<0.05). Conclusions NF κB modulated genes expression and protein synthesis of ICAM 1 and MCP 1; Celluar proliferation in vessel wall was dynamic change after balloon angioplasty injury; Antisense and decoy oligonucleotide of NF κB by local lipofectamine transfer inhibited NF κB activating genes modulation and the combined effect were remarkable than alone.

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周俊,陆国平,戚文航.核因子κB的反义和诱骗性寡核苷酸对大鼠球囊损伤后血管狭窄和新生内膜形成的影响[J].中国动脉硬化杂志,2003,11(4):314~318.

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  • 收稿日期:2002-09-20
  • 最后修改日期:2003-02-11
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