SYBR GreenⅠ荧光定量PCR检测肾移植患者CsA效用  

SYBR GreenⅠ fluoresence quantitative reverse transcriptase polymerase chain reaction for measurement of cyclosporine A effect in patients with renal transplantation

作　　者：王科[1] 王晓雄[1] 石炳毅[2] 钱叶勇[2] 贾金风[2] 常京元[2] 柏宏伟[2] 

机构地区：[1]解放军总医院泌尿外科,北京100853 [2]解放军总医院第309临床部,北京100853

出　　处：《军医进修学院学报》2006年第3期233-235,共3页Academic Journal of Pla Postgraduate Medical School

基　　金："十五"全军重大课题项目(01Z097)

摘　　要：目的:检测肾移植患者外周血淋巴细胞(PBL)有关细胞因子基因表达,判定环孢素A(CsA)效用,进一步验证CsA服用后2h浓度(C2)测定的必要性。方法:采用SYBR GreenⅠ荧光定量PCR对20例肾移植患者术后第7天PBL的IL-2和IFN-γmRNA表达进行检测,判定CsA的免疫抑制效用,对比CsA空腹浓度(C0)和C2与细胞因子抑制关系。结果:C2对应IL-2和IFN-γmRNA/β-actin mRNA相对模板数分别为0.038±0.006和0.018±0.004,C2对IL-2和IFN-γmRNA表达抑制率分别为(72.42±3.95)%和(69.52±6.97)%;C2与IL-2和IFN-γmRNA表达的抑制率有良好的线性关系(P<0.01);C0与IL-2和IFN-γmRNA表达的抑制率无良好的线性关系(P(0.05)。结论:SYBR GreenⅠ荧光定量PCR技术检测肾移植患者术后PBL的IL-2和IFN-γmRNA的表达,是判定CsA效用和研究免疫耐受的有效方法;C2仍为判定CsA生物利用度的重要指标。Objective： To measure expressions of cytokines genes in peripheral blood lymphocyte of patients after renal transplantation, evaluate cyclosporine A effect, further verify the necessity of testing cyclosporine A concentration 2. Methods： By SYBR Green Ⅰ PCR technique, we measured IL-2 and IFN-γ mRNA expressions of PBL from 20 cases of patients 7 days after renal transplantation, assessed immunosuppressive effect of cyclosporine A, contrasted the relation of CO and C2 with cytokines inhibiting. Results： Corresponding to C2, IL-2 and IFN-γ/ mRNA/β-actin mRNA relative templates were 0. 038 ± 0.006和10.018 ± 0.004 respectively, IL-2 and IFN-γ/inhibiting rates were （72.42 ± 3.95 ） % 和 （69.52 ± 6.97 ） %. linear correlation of C2 with inhibiting rate of IL-2,IFN-γ mRNA expression was fine（ P 〈 0.01 ）, but the one of CO with them was bad （P 〉0.05）. Conclusions： To measure IL-2 and IFN-γ mRNA expressions in PBLof patients after renal transplantation with SYBR Green Ⅰ PCR technique is an effective method to evaluate cyclosporine A effect and immunosuppressive study, C2 is still an important index to judge biological availability of cyclosporine A.

关 键 词：荧光定量反转录聚合酶链反应 肾移植 环孢素 

分 类 号：R699[医药卫生—泌尿科学]