转人α1,2岩藻糖苷转移酶基因猪动脉内皮细胞的活化抑制实验  

Human α1, 2-fucosyltransferase (HT) gene transferring is associated with inhibition of arterial endothelial cell activation in porcine xenograft models

作　　者：马志方[1] 王广有[1] 刘秉乾[1] 李胜芝[1] 张玥[1] 马腾骧[1] 

机构地区：[1]天津市泌尿外科研究所分子生物学教研室

出　　处：《中国药物与临床》2008年第1期22-25,共4页Chinese Remedies & Clinics

基　　金：天津市科委科技发展计划重点项目(043803411)

摘　　要：目的探讨人α1,2岩藻糖苷转移酶(HT)基因转移抑制猪动脉内皮细胞(PAEC)Ⅱ型活化的作用和机制。方法用不同条件刺激转人HT基因和正常的PAEC,在不同时段分别采用细胞酶联免疫吸附测定方法检测细胞间黏附分子(ICAM)-1在细胞表面的表达。采用免疫细胞化学方法检测细胞接受刺激12h后2种PAEC核因子(NF)-κB阳性细胞百分数。结果3种刺激均可以活化PAEC,使其表面ICAM-1的表达逐步升高,并在12h左右达到峰值。实验组和对照组细胞在接受刺激4h后ICAM-1的表达差异有统计学意义(P值均<0.05),实验组细胞ICAM-1的表达量均低于对照组,细胞的活化受到抑制。在不同条件下刺激12h后2种细胞的NF-κB阳性细胞百分数差异有统计学意义(P值均<0.05),表达人HT的PAECNF-κB阳性细胞百分数比正常猪动脉内皮细胞低。结论表达人HT的PAECⅡ型活化受到抑制,人HT基因转移可能一定程度抑制异种移植急性血管排斥反应(AVR),并且可能通过NF-κB发挥作用,PAECⅡ型活化的条件是多样的。Objective To explore the mechanism of human α1, 2-fueosyltransferase （HT） gene transferring in inhibiting type Ⅱ arterial endothelial cells （PAECs） activation in xenograft porcine models. Methods Transgenic and normal PAECs were stimulated with 5 % human serum, 5 % human serum of inactivated complement and 1000 U/ml human TNF-α, respectively. ICAM-1 expression on the PAEC was assayed by cell ELISA after 2, 4, 8, 12 and 24 h respectively. At 12 h, NF-kappa B expression in the two types of PAEC stimulated under three conditions was assayed by immunocytochemistry. Results The three stimulations were shown to activate PAECs, with ICAM-1 increasingly expressed on the PAEC over time and peaking after 12 h. After 4 h of stimulation onwards, transgenic PAECs showed less ICAM-1 expression on cell membrane as compared with normal PAECs （all P〈0.05）, which implicated inhibition of PAEC activation. Under all conditions, percentages of NF-kappa B positive cells were lower in transgenic than in normal PAECs （all P〈0.05）. Conclusions The type Ⅱ activation of transgenic PAEC expressing human HT was inhibited. To a certain degree, human HT gene transferring appears to suppress acute vascular rejection （AVR） where NF-kappa B may be playing an important role. In addition, type Ⅱ activation of endothelial cells can be muhifactorial.

关 键 词：移植 异种 基因转移技术 岩藻糖基转移酶类 内皮细胞 

分 类 号：R346[医药卫生—基础医学]