急性胰腺炎患者外周血单核细胞TLR9 mRNA的表达及意义

Expression of TLR9 mRNA in peripheral blood mononuclear cells of patients with acute pancreatitis

目的观察急性胰腺炎（AP）患者外周血单核细胞（PBMCs）Toll样受体9（TLR9）mRNA的表达情况。方法收集52例发病24h内的AP患者,采集第1、3、5天外周乙二胺四乙酸二钾（EDTAK2）抗凝静脉血,提取血浆低温冻存,用于成批检测胰弹性蛋白酶、促炎细胞因子、抗炎细胞因子等。同法采集其治愈后2~3个月的外周EDTAK2抗凝静脉血行上述检测,作为相应指标的基准水平值。分离外周血单核细胞用于随后成批作逆转录聚合酶链反应（RT-PCR）检测TLR9mRNA的表达变化规律。同法采集其治愈后3个月的外周EDTAK2抗凝静脉血行上述检测,作为相应指标的基准水平值。最终将成功采得治愈后3个月血液标本的前36例患者纳入最终研究。结果 AP患者的外周血PBMCs TLR9mRNA相对含量高于其痊愈后3个月的基线水平（P〈0.05）。AP患者的外周血PBMCs TLR9mRNA相对含量表达改变与胰弹性蛋白酶、促炎细胞因子水平呈正相关,与抗炎细胞因子无相关性。结论 AP患者的外周血PBMCs TLR9mRNA表达显著升高并与促炎细胞因子表达同步上调,提示AP的发生、发展可能通过TLR9介导。

Objective To investigate the expression of TLR9 mRNA of peripheral blood mononuclear cells（PBMCs）in patients with acute pancreatitis.Methods Fifty two AP patients with the disease duration in 24 hwere collected,peripheral EDTAK2 coagulation vein blood were collected on the first,third and fifth day,then plasma were cryop reserved to detect pancreatic elastase,proinflammatory cytokines and anti-inflammatory cytokines.Then the peripheral EDTAK2 coagulation vein blood two to three months after treatment were collected in the same method to undertake these tests,and act as the reference level value.Peripheral blood was collected from 36 acute pancreatitis patients.Three months later,peripheral blood was collected again from these 36 people as controls.PBMCs were isolated by Ficoll-Hypaque gradient centrifugation.RT-PCR was adopted to determine the relative content of the expression of TLR9 mRNA of the PBMCs.Results The relative content of expression of TLR9 mRNA were significantly up-regulated in the patients with acute pancreatitis,compared with that of controls（P〈0.05）.The up-regulated expression of TLR9 mRNA was related to expression of TNF-a and IL-1.Conclusion The up-regulated expression of TLR9 mRNA in acute pancreatitis patients indicates that infective factors might be mediated by TLR9.