转录因子维甲酸孤独受体γτ与刀叉样盒蛋白-3在肝囊型包虫病患者外周血中的mRNA表达及其意义

Quantitative gene expression of transcription factors RORγτ and FoxP3 in patients with liver cystic echinococcosis Turgun.

目的探讨维甲酸孤独受体γτ（RORγτ）和刀叉样盒蛋白-3（FoxP3）在肝囊型包虫病患者外周血中的mRNA的表达及其意义。方法将54例受试者分为3组：健康志愿者（HD）组20例，肝囊型包虫病（CE）组20例，肝囊型包虫病复发（RCE）组14例。应用实时荧光定量逆转录-聚合酶链反应（RT-PCR）法，检测各组受试者外周血单个核细胞中RORγτ和FoxP3的mRNA表达，并用t检验分析其结果。结果实时荧光定量RT-PCR检测RORγτ和FoxP3的扩增效率为90％-100％，批内和批间实验循环阈值（α）变化〈0．2。与HD组比较，RORγτ（RORγτ拷贝／GAPDH拷贝）在CE组和RCE组表达降低，差异有统计学意义（t=2．91，t=2．79，P〈0．05），而CE组和RCE组之间差异无统计学意义（t=0．56，P〉0．05）。FoxP3表达（FoxP3拷贝／GAPDH拷贝）在CE组和RCE组中较HD组增高，差异无统计学意义（t=0．49，t=0．02，P〉0．05）。RORγτ／FoxP3的比值在CE和RCE组中较HD组降低且差异有统计学意义（t=0．33，t=0．35，P〈0．01），而CE和RCE组之间差异无统计学意义（t=0．48，P〉0．05）。结论实时荧光定量RT-PCR检测RORγτ和FoxP3的mRNA表达结果准确可靠。在肝囊型包虫病患者外周血单个核细胞中，FoxP3表达升高，RORγτ/FoxP3表达且以降低RORγτ表达降低起主导作用。这可能与肝包虫感染过程中的免疫逃避现象有关。RORγτ／FoxP3的比值比单独检测RORγτ或FoxP3更能客观评价肝囊型包虫病感染过程中的免疫逃避状态。

Objective To investigate the mRNA expression and significance of transcription factors RORγτ and FoxP3 in peripheral mononuclear cells in patients with liver cystic echinococcosis, nethotis Ffity-four subjects were enrolled and derided into three groups： ealthy donor （HD, n = 20）, liver cystic echinococcosis （CE,n = 20）, ecuured cystic echinococcosis （RCE,n = g）. The expression of transcrition factors RORγτ and FoxP3 mRNA in peripheral mononuclear cells of patients with liver cystic echinococcosis was detected by real-time polymerase chain reaction （PCR）. Results The amplification efficiency of quantification of RORγτ and FoxP3 with real-time PCR was 90% -100%. The Ct in intra- and interassay was 〈 0. 20. As compared with HD group, the expression of RORγτ（ RORγτ copies/GAPDH copies） was decreased in CE and RCE groups （ t = 2. 91, t = 2. 79, P 〈 0.05 ） , but there was no statistically significant difference between CE and RCE groups （ t = 0. 56, P 〉 0.05 ）. Oppositely, the expression of FoxP3 （FoxP3 copies/GAPDH copies） was increased in CE and RCE groups as compared with HD group, but there was statistically significant difference （ t = 0.49, t = 0. 02, P 〉 0. 05 ）. The ratio of RORγτ and FoxP3 was obviously decreased in CE and RCE groups as compared with HD group （ t = 0. 33, t = 0. 35, P 〈0. 01 ）, but there was no significant difference between CE and RCE groups （ t = 0. 48, P 〉 0. 05 ）. Conclusion The real-time PCR assay for quantification of RORγτ and FoxP3 is efficient, special and productive. The increased expression of FoxP3 and decreased expression of RORγτ and ROR3,＇r/FoxP3 are related to special immunity during the pathogenesis of liver cystic echinococcosis, The decreased expression of RORγτ may play a major role and the RORγτ/FoxP3 expression ratio may be used as a more objective marker than RORγτ and FoxP3 alone for estimation of special immunovasion during the infection.