CAJ | 학술논문

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(Ct)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和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.
목적 탐토유갑산고독수체γτ(RORγτ)화도차양합단백-3(FoxP3)재간낭형포충병환자외주혈중적mRNA적표체급기의의.방법 장54례수시자분위3조:건강지원자(HD)조20례,간낭형포충병(CE)조20례,간낭형포충병복발(RCE)조14례.응용실시형광정량역전록-취합매련반응(RT-PCR)법,검측각조수시자외주혈단개핵세포중RORγτ화FoxP3적mRNA표체,병용t검험분석기결과.결과 실시형광정량RT-PCR검측RORγτ화FoxP3적확증효솔위90%～100%,비내화비간실험순배역치(Ct)변화＜0.2.여HD조비교,RORγτ(RORγτ고패/GAPDH고패)재CE조화RCE조표체강저,차이유통계학의의(t=2.91,t=2.79,P＜0.05),이CE조화RCE조지간차이무통계학의의(t=0.56,P＞0.05).FoxP3표체(FoxP3고패/GAPDH고패)재CE조화RCE조중교HD조증고,차이무통계학의의(t=0.49,t=0.02,P＞0.05).RORγτ/FoxP3적비치재CE화RCE조중교HD조강저차차이유통계학의의(t=0.33,t=0.35,P＜0.01),이CE화RCE조지간차이무통계학의의(t=0.48,P＞0.05).결론 실시형광정량RT-PCR검측RORγτ화FoxP3적mRNA표체결과준학가고.재간낭형포충병환자외주혈단개핵세포중,FoxP3표체승고,RORγτ화RORγτ/FoxP3표체차이강저RORγτ표체강저기주도작용.저가능여간포충감염과정중적면역도피현상유관.RORγτ/FoxP3적비치비단독검측RORγτ혹FoxP3경능객관평개간낭형포충병감염과정중적면역도피상태.
Objective To investigate the mRNA expression and significance of transcription factors RORγτ and FoxP3 in peripheral mononuclear cells in patients with liver cystic echinococcosis. Methods Ffity-four subjects were enrolled and devided into three groups: ealthy donor ( HD, n = 20), liver cystic echinococcosis ( CE,n =20), ecuured cystic echinococcosis ( RCE,n = 14). 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 RORγτ/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 immuno-evasion during the infection.