摘要
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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2011年第1期72-75,共4页
Chinese Journal of Experimental Surgery
关键词
肝囊型包虫病
T细胞
转录因子
免疫逃避
Liver echinococcosis
T cells
Transcription factor
Immune evasion