铜绿假单胞菌影响外周血单个核细胞主要组织相容性复合物的实验研究

The expression of peripheral blood monouclear MHC affected by Pseudomonas aeruginosa

目的研究铜绿假单胞菌对外周血单个核细胞主要组织相容性复合物(MHC)基因表达的影响。方法 10只新西兰白兔随机分成2组,实验组6只,对照组4只。实验组6只新西兰白兔于实验前1天皮下注射铜绿假单胞菌,每2 d无菌抽取实验兔外周血2 mL,乙二胺四乙酸二钾(EDTA-K_2)抗凝,先检测血常规,剩余抗凝血用于分离外周血单个核细胞,逆转录-聚合酶链反应(RT-PCR)测定主要组织相容性复合物(MHC)Ⅰ类和MHC-Ⅱ类mRNA的表达,蛋白印迹法测定其蛋白表达,实验持续8d。结果第8天实验结束时,实验组外周血单个核细胞MHC-Ⅰ类、MHC-Ⅱ类mRNA的表达由实验前的0.65±0.05和2.38±0.25分别升高到3.51±0.42和6.51±0.52,蛋白表达由实验前的2.63±0.05和12.38±1.27分别升高到13.52±0.82和31.23±2.82。结论铜绿假单胞菌能明显升高外周血单个核细胞MHC基因的表达,从而可能升高移植脏器的MHC表达,诱发急性移植排斥反应。

Objective To study peripheral blood monouclear MHC gene expression affected by pseudomonas aeruginosa. Methods Ten New Zealan white rabbits were randomly divided into two groups,four rabbits served as the control group,six rabbits in experimental group. The experiment group subcutaneous injected pseudomonas aeruginosa one day before experiment, rabbits blood sample were drawn every two days,at first the blood routine were detected, then obtained peripheral blood mononuclear cells from remainder anticoagulative blood,RT-PCR and Western blotting were used to examine the mRNA expression and protein expression of MHC I and MHC II , the experiment lasted eight days. Results When the experiment were end in day 8, peripheral blood monouclear MHC I and MHC II mRNA expression in experiment group were 3.51 ± 0.42 and 6.51 ±0.52, both significantly higher than those before the experiment （0.65±0.05 and 2.38±0.25,both P〈0.01）, MHC I and MHCII protein expression in experiment group were 13.52±0.82 and 31.23±2.82 ,both significantly higher than those before the experiment（2.63±0.05 and 12.38±1.27,both P〈0.01）. Conclusion Pseudomonas aeruginosa can significantly increase peripheral blood monouelear MHC gene expression,so maybe increase graft organ MHC gene expression,induced acute graft rejection.