Early detection of postoperative sepsis and organ dysfunction using cDNA micro-array and therapeutic strategy for these complications by anti-TLR antibody.

背景/目的：积极的肝胆胰手术与高并发症发生率相关。应用cDNA微阵列和转录聚合酶链反应（RT-PCR）对肝胆胰手术患者术后脓毒症和脏器功能障碍的早期检测进行评价。此外，在腹膜败血症或梗阻性黄疸动物模型中测定抗tlr2、4抗体对这些并发症的治疗效果。方法：采用ELISA法测定56例连续行肝胆胰手术患者围手术期血浆白细胞介素(IL)-6、IL-10、可溶性淋巴细胞活化基因(sLAG) -3、可溶性CD30 （sCD30）浓度。采用cDNA微阵列技术分析PBMCs的基因表达，采用逆转录聚合酶链反应（RT-PCR）检测T-bet、GATA-3、TLR2和TLR4 mRNA在PBMCs中的表达水平。在腹膜脓毒症和梗阻性黄疸动物模型中观察抗tlr2、4抗体对脓毒症的影响。结果：56例患者中，术后感染30例。术后感染患者血浆IL-6、IL-10水平明显高于未感染患者（p<0.05）。术后感染患者sCD30水平术前明显升高，术后第7天sCD30水平继续升高（p<0.05）。两组间sLAG-3水平无显著差异。术后感染患者GATA-3 mRNA表达水平术前明显升高，术后第7天仍保持较高水平（p<0.05）。术后感染患者术后第3、7、14天T-bet mRNA表达水平显著升高（p<0.05）。术后感染患者TLR2和TLR4 mRNA表达水平在术后第3天显著升高（p< 0.05）。术前GATA-3 mRNA表达水平与TLR2、TLR4 mRNA表达水平显著相关（p<0.05）。抗tlr2、4抗体对腹膜败血症或梗阻性黄疸动物模型无效。结论：这些结果提示术前Th1/Th2平衡在术后感染患者中向Th2优势转移，术前外周血sCD30和GATA3 mRNA表达水平是预测术后脓毒症和器官功能障碍的新标志物。少

Background/Objectives : Aggressive hepatobiliary pancreatic surgery has been associated with high complication rates. Early detection of postoperative sepsis and organ dysfunction using cDNA micro-array and transcription-polymerase chain reaction (RT-PCR) was evaluated in patients undergoing hepatobiliary pancreatic surgery. Furthermore, therapeutic effects of anti-TLR2, 4 antibodies on these complications were determined in animal models with peritoneal sepsis or obstructive jaundice.Methods : Plasma concentrations of interleukin (IL)-6, IL-10, soluble lymphocyte activation gene (sLAG) -3, and soluble CD30 (sCD30) were determined by ELISA perioperatively in consecutive 56 patients who underwent hepatobiliary pancreatic surgery. Gene expression in PBMCs was analysed by cDNA micro-array, and expression levels of T-bet, GATA-3, TLR2, and TLR4 mRNA in PBMCs were confirmed by reverse RT-PCR.Effects of anti-TLR2, 4 antibodies on sepsis were determined in animal models with peritoneal sepsis … More or obstructive jaundice.Results : Of the 56 patients, 30 patients had postoperative infection. Postoperative plasma levels of IL-6 and IL-10 were significantly higher in patients with postoperative infection than in those without infection (p<0.05). sCD30 levels were significantly higher preoperatively, and remained higher by postoperative day 7 in patients with postoperative infection (p<0.05). sLAG-3 levels were not significantly different between two groups. GATA-3 mRNA expression level was significantly higher preoperatively, and remained higher by postoperative day 7 in patients with postoperative infection (p<0.05). T-bet mRNA expression level was significantly higher on postoperative days 3, 7, and 14 in patients with postoperative infection (p<0.05). TLR2 and TLR4 mRNA expression levels were significantly higher on postoperative day 3 in patients with postoperative infection (p< 0.05). Preoperative expression levels of GATA-3 mRNA correlated significantly with those of TLR2 and TLR4 mRNA (p<0.05). Anti-TLR2, 4 antibodies were not effective in animal models with peritoneal sepsis or obstructive jaundice.Conclusions : These results suggest that preoperative Th1/Th2 balance shifts toward Th2 dominance in patients with postoperative infection, and preoperative expression levels of circulating sCD30 and GATA3 mRNA in PBMCs are the novel markers predicting postoperative sepsis and organ dysfunction. Less