CAJ | 학술논문

目的探讨microRNA(即miRNA)在大鼠肝移植急性排斥反应中的表达情况.方法以Kamada“双袖套”法行大鼠原位肝移植术(ROLT),分为排斥组(Lewis/BN,n=12)、无排斥组(Lewis/Lewis,n=12).术后第7d采用miRNA芯片技术确定外周血miRNA表达谱,采用生物信息学方法筛选出表达差异显著的特定miRNA,进而利用qRT-PCR技术对ROLT术后第3、5、7d该特定miRNA进行定量分析,同时结合病理、肝功能结果进行动态化对比分析.结果 miR-206在排斥组与无排斥组的表达差异最为显著.排斥组外周血miR-206表达从术后第3d既显著高于无排斥组(t=8.875,P=0.001),且表达量随术后时间延长而逐步上升(F=32.154,P＜0.01).而两组的排斥反应活动指数(RAI评分)到术后第5d、肝功能指标(TBil、ALT)到术后第7d才出现显著差异.外周血和移植肝中miR-206表达量与RAI评分呈强正相关(rs分别为0.812及0.881,P＜0.01).在排斥组,外周血与移植肝中miR-206表达量呈正相关(rp=0.644,P=0.024).结论对于大鼠肝移植急性排斥反应的诊断,外周血特定miRNA检测具有早期、灵敏、特异、稳定等优势,是一种优于病理检查和肝功能化验的诊断方法.
목적 탐토microRNA(즉miRNA)재대서간이식급성배척반응중적표체정황.방법 이Kamada“쌍수투”법행대서원위간이식술(ROLT),분위배척조(Lewis/BN,n=12)、무배척조(Lewis/Lewis,n=12).술후제7d채용miRNA심편기술학정외주혈miRNA표체보,채용생물신식학방법사선출표체차이현저적특정miRNA,진이이용qRT-PCR기술대ROLT술후제3、5、7d해특정miRNA진행정량분석,동시결합병리、간공능결과진행동태화대비분석.결과 miR-206재배척조여무배척조적표체차이최위현저.배척조외주혈miR-206표체종술후제3d기현저고우무배척조(t=8.875,P=0.001),차표체량수술후시간연장이축보상승(F=32.154,P＜0.01).이량조적배척반응활동지수(RAI평분)도술후제5d、간공능지표(TBil、ALT)도술후제7d재출현현저차이.외주혈화이식간중miR-206표체량여RAI평분정강정상관(rs분별위0.812급0.881,P＜0.01).재배척조,외주혈여이식간중miR-206표체량정정상관(rp=0.644,P=0.024).결론 대우대서간이식급성배척반응적진단,외주혈특정miRNA검측구유조기、령민、특이、은정등우세,시일충우우병리검사화간공능화험적진단방법.
Objective To investigate the role of miRNA expression in the acute rejection of liver transplantation in rats.Methods The rat orthotopic liver transplantation (ROLT) was performed using the two-cuff technique described by Kamada with modification.Rats were randomly divided into two groups,a rejection group (n=12) and a control group (n=12).MicroRNA microarray technology was used to screen the microRNA expression profile in peripheral blood on the 7th postoperative day,and the bioinformatics microRNA screening method detected that miR-206 had the biggest difference in expression level.Peripheral blood samples were gathered before,3 days,5 days,and 7 days after ROLT to analyze the expression level of miR 206 quantitatively with qRT PCR technology.Dynamic contrast analysis was made between the quantitative results of miR-206 and the results of pathological examination and liver function tests.Results There was no significant difference in miR-206 expression level between the rejection group and control group before the operation (t=0.105,P=0.921).MiR-206 expression was significantly higher in the rejection group than in the control group 3 days after ROLT (t=8.875,P=0.001).The expression level of miR 206 in the rejection group also increased gradually over time (F=32.154,P＜0.01).However,pathological changes and liver function index did not show obvious differences until the 5th and 7th day postoperatively.The expression of miR-206 in peripheral blood and graft was strongly and positively correlated with the rejection active index (RAI) of acute liver rejection pathology (0.812 and 0.881 respectively,P＜0.01).In the rejection group,the miR-206 level in peripheral blood and graft were positively associated with each other.Conclusions Detection of miR-206 in peripheral blood was significantly better than pathological examination and liver function test in the diagnosis of acute rejection of rat liver transplantation.Moreover,miR-206 was an early sensitive and specific marker for making this diagnosis.