核因子κB诱骗寡核苷酸对大鼠肝脏的Kupffer细胞共刺激分子表达的影响  

The effects of NF-κB decoy oligodeoxynucleotides on expression of co-stimulatory molecules in Kupffer cells

作　　者：李敬东[1] 刘海忠[2] 刘作金[2] 龚建平[2] 肖江卫[1] 彭勇[1] 

机构地区：[1]川北医学院附属医院普外三科,四川南充637000 [2]重庆医科大学附属第二医院肝胆外科

出　　处：《中华器官移植杂志》2009年第3期137-140,共4页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金（30300337,30700773）

摘　　要：目的探讨核因子κB（NF-κB）诱骗寡核苷酸（ODN）对大鼠肝脏Kupffer细胞（KC）膜表面共刺激分子表达的影响。方法取雄性SD大鼠，麻醉后经门静脉插管，以0．5g／LIV型胶原酶体外循环灌注消化肝脏，不连续Percoll密度梯度离心分离KC。以人工合成的NF-κB诱骗ODN转染KC，再以终浓度为img／L的脂多糖（LPS）刺激（NF-κB诱骗组），培养8h后，采用逆转录聚合酶链反应测定KC膜表面CD80、CD40和CD54mRNA的表达。以不转染NF-xB诱骗ODN、但接受LPS刺激的KC为对照（LPS刺激组），正常大鼠KC为对照组。每组每份标本含KC1×10^5个。结果每个大鼠肝脏可以分离得到KC（3-4）×10^6个，NF-κB诱骗ODN可以在KC内维持12d而不被降解。对照组KC膜表面CD80、CD40、CD54 mRNA呈低表达；LPS刺激组KC表面3种共刺激分子表达均较对照组明显升高，分别是对照组的2．53、2．51和2．74倍（P〈0．01）；NF-κB诱骗组CD80和CD40mRNA表达明显低于LPS组，仅为LPS组的0．52和0．55倍（P〈0．01），而CD54 mRNA表达的差异无统计学意义（P〉0．05）。结论在体外，NF-κB诱骗ODN可以高效地抑制KC表面共刺激分子的表达，为活体内应用NF-κB诱骗ODN抑制肝移植后急性排斥反应提供了实验依据。Objective To investigate the inhibitory effects of NF-κB decoy oligodeoxynucleotides （ODN） on the expression of co-stimulatory molecules in Kupffer cells （KCs）. Methods KCs were isolated by in situ collagenase perfusion and randomly divided into three groups： control group, LPS stimulation group （ 1 mg/L LPS）, and NF-κB decoy ODN group, in which KCs were transfected with NF-κB decoy ODN （4μg/1 × 10^5 KCs） before LPS stimulation. After 8 h of LPS stimulation, KCs were collected and CD80,CD40, and CD54 mRNA expression was assayed by reverse transcription-polymerase chain reaction. Results LPS stimulation significantly increased the expression of co-stimulatory molecules （CD80, CD40, and CD54） in KCs by 2. 53,2. 51, and 2. 74 fold respectively over that of the control group （P〈0. 01）. NF-κB decoy ODN could efficiently suppress KCs activation. In decoy group, the expression of CD80 and CD40 mRNA was significantly decreased by 0. 52 and 0. 55 fold respectively over that of the LPS group （P〈0. 01） ,while that of CD54 mRNA was not （P）0. 05 ）. Conclusion NF-κB decoy ODN can efficiently inhibit the expression of co-stimulatory molecules in KCs,which provides reliable experimental data for the clinical application of NF-κB decoy ODN to inhibit acute rejection after liver transplantation in vivo.

关 键 词：NF-κB 寡核苷酸类 枯否细胞 抗原 CD80 抗原 CD40 

分 类 号：R686[医药卫生—骨科学]