CAJ | 학술논문

目的探讨T-bet(T-box expression in T cells)和GATA结合蛋白3(GATA-3)在大鼠肺移植急性排斥反应中的mRNA表达及其意义.方法建立同种异体大鼠肺移植模型,实验分为正常对照组(n=5只)、同种异体移植3 d组(n=4只)和5 d组(n=6只),用实时荧光定量逆转录-聚合酶链反应(RT-PCR)法,检测移植后肺组织中T-bet和GATA-3的mRNA表达.结果实时荧光定量RT-PCR检测T-bet和GATA-3的扩增效率为78%～85%,批内和批间实验循环阈值(Ct)变化均＜0.15.与止常对照和右侧非移植肺比较,同种异体移植术后3 d组,T-bet和GATA-3无显著性改变,但其比值明显高于右侧肺;同种异体移植术后5 d组,T-bet表达增高(9.31拷贝/106 18 S拷贝),但差异无统计学意义(P＞0.05),GATA-3表达下降(0.88拷贝/105 18 S拷贝),T-bet/GATA3的比值增高(1.12),差异均有统计学意义(P＜0.05).结论实时荧光定量RT-PCR检测T-bet和GATA-3结果准确可靠.在急性排斥反应中,T-bet表达升高,GATA-3表达降低,其中GATA-3的表达起主导作用.T-bet/GATA-3的比值比单独检测T-bet或GATA-3更能客观地评价大鼠肺移植急性排斥反应.
목적 탐토T-bet(T-box expression in T cells)화GATA결합단백3(GATA-3)재대서폐이식급성배척반응중적mRNA표체급기의의.방법 건립동충이체대서폐이식모형,실험분위정상대조조(n=5지)、동충이체이식3 d조(n=4지)화5 d조(n=6지),용실시형광정량역전록-취합매련반응(RT-PCR)법,검측이식후폐조직중T-bet화GATA-3적mRNA표체.결과 실시형광정량RT-PCR검측T-bet화GATA-3적확증효솔위78%～85%,비내화비간실험순배역치(Ct)변화균＜0.15.여지상대조화우측비이식폐비교,동충이체이식술후3 d조,T-bet화GATA-3무현저성개변,단기비치명현고우우측폐;동충이체이식술후5 d조,T-bet표체증고(9.31고패/106 18 S고패),단차이무통계학의의(P＞0.05),GATA-3표체하강(0.88고패/105 18 S고패),T-bet/GATA3적비치증고(1.12),차이균유통계학의의(P＜0.05).결론 실시형광정량RT-PCR검측T-bet화GATA-3결과준학가고.재급성배척반응중,T-bet표체승고,GATA-3표체강저,기중GATA-3적표체기주도작용.T-bet/GATA-3적비치비단독검측T-bet혹GATA-3경능객관지평개대서폐이식급성배척반응.
Objective To investigate the mRNA expression and significance of transcription factots T-bet and GATA-3 in rat lung acute allograft rejection.Methods Experimental animals were established and divided into normal group(n=5),allogeneic 3-day group(n=4)and allogeneic 5-day group (n=6).The expression of transcription factors T-bet and GATA-3 mRNA in rat lung allografts was detected by real-time PCR.Results The amplification efficiency of quantification of T-bet and GATA-3 with real-time PCR was 78%-85%.The Ct in intra-and interassay was＜0.15.Compared with normal controls (n=5),the expression had no significant change in allogeneic 3-day group:the expression of T-bet was increased(9.60±4.07 copies/106 copies),the expression of GATA-3 was decreased(0.82±0.04 copies/106 copies),and the ratio of T-bet and GATA-3 was increased(1.26±0.40 copies/106copies)in allogeneic 5-day group(P＜0.05).Conclusion The real-time PCR assay for quantification of T-bet and GATA-3 mRNA is efficient.special and reproductive.The increased T-bet and decreased GATA-3 are relative to acute rejcction.The expression of GATA-3 plays a maior role in acute rejection.Quantification of the ratio of T-bet/GATA-3 can be used as a more objective marker than T-bet or GATA-3 alone for estimation of the acute rejection of lung transplantation.