黏附分子__(期刊论文)

脂质体介导NFκB decoy ODN低温缺氧对ECV304的转染及复氧后对其黏附分子mRNA表

http://www.100md.com 《第四军医大学学报》 2004年第15期

黏附分子,,内皮细胞株ECV304；低温低氧；脂质体；转染；黏附分子,脂质体介导NFκBdecoyODN低温缺氧对ECV304的转染及复氧后对其黏附分子mRNA表达的影响,0引言,1材料和方

     Transfection of NFκB decoy ODN mediated by liposome on ECV304 under hypothermic/anoxic condition and influence on expression of adhesive molecules after reoxygenation

    FENG GuiWen, YU LiXin, ZHAO XianGuo, ZHANG HongTao

    1Department of Kidney Transplantation, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China, 2Center of Hemodialysis, First Hospital, Zhengzhou University, Zhengzhou 450052, China

    【Abstract】 AIM: To investigate the transfection efficiency of NFκB decoy ODN mediated by liposome on ECV304 cells in EuroCollin’s (Ecs) under hypothermic and anoxic condition and the influences on its NFκB activation and the expression of proinflammatory molecules in ECV304 after reoxygenation. METHODS: ① ECV304 monolayers were transfected with liposome/decoy ODN or scrambled ODN complexes in ECs under hypothermic and anoxic condition. ② The transfection efficiency was evaluated by flow cytometry and the distribution of FITClabeled decoy ODN in ECV304 cells was observed by fluorescence microscope. ③ The NFκB activation of ECV304 at different period after reoxygenation was analyzed by electrophoretic mobility shift assay (EMSA). ④ Genes expression of ICAM1, VCAM1 and Pselectin was assessed by reverse transcription polymerase chain reaction (RTPCR). RESULTS: MFI of Liposome/decoy ODN group was 1072.7±33.1, 91.1 folds of naked ODN group, and the transfection efficiency of Liposome/decoy ODN group was (93.1±2.6)%, 71.5 folds of naked ODN group. Intensive fluorescence was seen in cell nuclei in liposome/decoy ODN group, but it was rarely seen in cell nuclei in naked ODN group. The NFκB activation of ECV304 at 1 h started rising and peaked at 4 h after reoxygenation. The NFκB activation of ECV304 transfected with decoy ODN at 4 h after reoxygenation was suppressed, but that of ECV304 transfected with scrambled ODN under the same condition was not suppressed. The gene expression of ICAM1, VCAM1 and Pseletin was significantly inhibited by NFκB decoy ODN. CONCLUSION: Decoy ODN can be transfected with high efficiency into ECV304 cells using in vivo liposome in ECs under hypothermic and anoxic condition while the naked ODN can not. The transfection of NFκB decoy ODN results in the inhibition of NFκB activation and genes expression encoding adhesive molecules in ECV304 cells. ......

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