急性胰腺炎患者循环miR-29a水平升高且与疾病严重程度及预后差正相关

Up-regulation of circulating miR-29a in patients with acute pancreatitis and is positively correlated with disease severity and poor prognosis

目的明确循环miR-29a在急性胰腺炎(AP)的诊断与监控预后中的应用价值。方法募集轻度急性胰腺炎(MAP30例、中度重症胰腺炎与重度急性胰腺炎(M-SAP)共30例,30例健康成人为对照组。比较一般临床指标,采用实时定量PCR检测外周血miR-29a水平并分析与临床评分的相关性。受试者工作特征(ROC)曲线评估miR-29a对AP的诊断价值。以脱氧胆酸(DCA)处理AR42J细胞建立AP细胞模型,Western blot法检测miR-29a及胱天蛋白酶3(caspase-3)表达量,采用慢病毒载体转入miR-29a模拟物或抑制物,Western blot法检测miR-29a及DCA处理后caspase-3的蛋白水平。结果 MAP组与M-SAP组急性期循环miR-29a相对表达量显著升高且M-SAP组高于MAP组,同组内恢复期miR-29a表达量低于急性期,且miR-29a表达量与Ranson评分及APACHEⅡ评分呈正相关。miR-29a用以诊断MAP的受试者工作特征曲线下面积(AUC)为0. 961 (95%CI:0. 921～1. 002),截断点(cut-off)值为1. 460;用以诊断M-SAP的AUC为0. 972 (95%CI:0. 939～1. 004),cut-off值为1. 340。AP细胞模型中,miR-29a与caspase-3水平升高;与空载体对照组相比,过表达miR-29a后caspase-3表达量升高,抑制miR-29a后则caspase-3水平降低。结论 MAP与M-SAP患者循环miR-29a水平明显升高,miR-29a高表达可能与胰腺腺泡细胞凋亡有关,该指标对于AP的诊断与预后监测具有较高的应用价值。

Objective To investigate the clinical application value of circulating miR-29 a in diagnosis and prognosis monitoring in acute pancreatitis( AP) patients,and to preliminary explore the molecular pathology significance of high expression ofmiR-29 a. Methods 30 cases of mild acute pancreatitis( MAP) patients( MAP group) and 30 cases of moderately serveacute pancreatitis( MSAP) or serve acute pancreatitis( SAP) patients( M-SAP group) were randomly selected,and 30 cases of healthy adults were used as control group. The general clinical data of each group were compared,and miR-29 alevels in peripheral blood were measured by real tome quantitative PCR,then the correlation between miR-29 a levels andRanson score or APACHEⅡscore were analyzed. The clinical usefulness of miR-29 a for AP patients was assessed by ROCcurve analysis. AR42 J cells were treated with deoxycholic acid( DCA) to establish AP cell model,the expression levels ofmiR-29 a and caspase-3 were detected. AR42 J cells were transfected by lentiviral vector contain miR-29 a mimic or miR-29 aAMO and measured of expression levels of miR-29 a and caspase-3. Results In acute phase,the expression of circulatingmiR-20 a was up-regulated in MAP and M-SAP group patients,and miR-20 a levels of M-SAP group patients were higher thanMAP group patients. In same group,miR-20 a levels in acute phase were higher than recovery phase. Correlation analysisindicated the positive correlation between miR-20 a levels and Ranson score or APACHE Ⅱ score. ROC curve analysisindicated that the AUC of miR-29 a for diagnosis MAP patients was 0. 961( 95% CI: 0. 921 ~ 1. 002),cut-off value was 1. 460.The AUC of miR-29 a for diagnosis M-SAP patients was 0. 972( 95% CI: 0. 939 ~ 1. 004),cut-off value was 1. 340. Theexpression levels of miR-29 a and caspase 3 were increased in AP cell model. Moreover,caspase 3 levels in AP cell modelwere increased than vector control after overexpression of miR-29 a,and caspase-3 levels were reduced than vector controlafter suppressing of miR-29 a expression. Conclusion Circulating miR-29 a is high expression in MAP and M-SAP patients,and high expression of miR-29 a may relate to pancreatic acinar cell apoptosis,and this biomarkers has high clinical value inAP diagnosis and prognosis monitoring.