中文摘要：

目的研究miR-146a对原发性胆汁性肝硬化（PBC）患者T细胞增殖以及白细胞介素（IL）-2分泌的调控作用。方法荧光定量聚合酶链反应（RT-PCR）检测20例PBC患者和20名健康个体外周血单个核细胞（PBMC）、辅助性T细胞、细胞毒性T细胞、B细胞和单核细胞内miR-146a的表达。将miR-146a表达载体和表达抑制体转染入PBC患者辅助性T细胞内，以抗CD3抗体和抗CD28抗体刺激辅助性T细胞，以CCK-8法检测辅助性T细胞增殖能力，酶联免疫吸附试验（ELISA）法检测培养基内IL-2水平，以RT．PCR法监测辅助性T细胞内miR．146a的表达。2组间的比较采用t检验。结果PBC患者PBMC内miR-146a的表达（0．46±0．20）较健康个体（1．00±0．26）明显降低（t=7．47，P〈0．01）。PBC患者辅助性T细胞和单核细胞内miR-146a的表达（分别为（0．33±0．13，0．56±0．11）较健康个体（分别为1．00±0．14，1．00±0．11））明显降低（t=6．15，4．97；P〈0．01或P〈0．05），但PBC患者与健康个体细胞毒性T细胞（0．98±0．15，1．00±0．12，t=0．22，P〉O．05）和B细胞（0．91±0．06，1．00±0．14；t=0．97，P〉0．05）内miR-146a的表达量之间的差异无统计学意义。抗C03和抗CD28可以诱导辅助性T细胞表达miR-146a（刺激前：1．00±0．18；刺激后：9．12±2．05；t=8．81，P〈0．01kPBC患者辅助性T细胞较健康个体具有较强的增殖能力{PBC：0．35±0．06[吸光度（A值）]；健康个体：0．26±0．04（A值）；t=2．83，P〈0．05}以及释放IL-2[PBC（686±109）pg/ml；健康个体（512±72）pg／ml；t=2．96，P〈0．05]的能力。miR-146a可以抑制PBC患者T细胞的增殖以及分泌IL-2的能力。结论miR-146a可以负向调控活化的PBC患者辅助性T细胞增殖以及分泌IL-2的能力。PBC的发病机制可能与患者辅助性I生T细胞内miR-146a表达降低、对细胞增殖的抑制作用减弱有关。

英文摘要：

Objective To investigate the effect of miR-146a on the proliferation and interleukin （IL）-2 production of T helper cells from primary biliary cirrhosis （PBC） patients. Methods MiR-146a in the peripheral blood mononuclear cells （PBMC）, monocytes, T helper cells, cytotoxic T cells and B ceils from 20 confirmede PBC patients and age/sex matched healthy controls were detected by quantitative PCR. By gainand-loss of function, the miR-146a＇s effect on anti-CD3/anti-CD28 activated T helper＇s proliferation and IL-2 production ability were measured by CCK-8 approach and enzyme linked immunosorbent assays（ELISA）, respectively. Statistical analysis were carried out by t-test. Results PBMCs （0.46±0.20 vs 1.00±0.26; t=7.47,P〈0.01）, T helpers （0.33±0.13 vs 1.00±0.14; t=6.15, P〈0.01） and monocytes （0.56±0.11 vs 1.00±0.11; t= 4.97, P〈0.05）, hut not B cells （0.91±0.06 vs 1.00±0.14; t=0.97, P〉0.05） and cytotoxic T cells （0.98±0.15 vs 1.00±0.12; t=0.22; P〉0.05） from PBC patients had lower miR-146a expression level than that of healthy controls. Inducible up expression of miR-146a was observed in PBC patients＇T helpers stimulated with anti- CD3/anti-CD28 （1.00±0.18 vs 9.12±2.05; t=8.81; P〈O.O1）. The activated T helpers from PBC patients had higher proliferative ability EPBC ： 0.35±0.06 （A） ; healthy controls ： 0.26±0.04 （A） ; t=2.83 ; P〈0.05 ] and increased IL-2 production [PBC： （685.60±109.19 pg/ml）]; Healthy controls： [（512.20±72.26） pg/ml; t= 2.96 ;pg /ml〈0.05 ] than those of healthy controls. For activated T helpers, the proliferation ability, as well as IL-2 production, was enhanced by miR-146a. Conclusion MiR-146a can clown regulate the proliferation and IL-2 releasing of activated T helpers. The reduced miR-146a expression enhances IL-2 production and promotes proliferation of T helper of PBC patients, thus, may be involved in the pathogenesis of PBC.