慢加急性肝衰竭患者Toll样受体3触发后DC分泌细胞因子的变化

目的探讨慢加急性肝衰竭（ACLF）患者外周血单核细胞衍生的树突状细胞（MoDC）上TLR3触发后细胞因子的分泌情况。方法选择ACLF患者60例,慢性乙型肝炎（CHB）患者40例,20例健康者作对照组（NC组）,取其抗凝全血,通过密度梯度法离心及免疫磁珠阳性选择法分离单核细胞,体外诱导培养为MoDC,经poly（I︰C）刺激后,实时荧光定量PCR检测TLR3及核因子κB和干扰素调节因子3的表达,液相蛋白流式检测促炎性因子IL-6、IL-8、IL-10、IL-12p70、IL-1β和TNF-α的分泌水平,均数间两两比较采用t检验。结果 poly（I︰C）刺激后,ACLF组患者NF-κBmRNA显著高于CHB组和NC组（P〈0.01）,而其IRF3mRNA低于CHB和NC组（P〈0.01）。3组研究对象MoDC分泌的IL-6及TNF-α差异有统计学意义（P〈0.005）,其分泌水平为ACLF组〉CHB组〉NC组。分泌的IL-10在ACLF组低于CHB组（P=0.021）,但与NC组差异无统计学意义。IL-12p70的分泌水平在ACLF组显著低于CHB组和NC组（P〈0.001）。IL-8分泌水平为ACLF组

Expression of cytokines in monocyte-derived dendritic cells after Toll like receptor 3 being triggered in patients with hepatitis B related acute-on-chronic liver failure

Objective To investigate the expression of cytokines in monocyte-derived dendritic cells (MoDC) after Toll like receptor (TLR) 3 being triggered in acute-on-chronic liver failure (ACLF) patients. Methods Peripheral blood was collected from 60 ACLF patients, 40 chronic hepatitis B (CHB) patients, and 20 healthy individuals who are served as normal controls (NC). Purified monocytes were isolated by combination of Histopaque-1.077 and CD14 Microbeads. MoDCs were induced from CD14+ monocytes and stimulated such cells in vitro with poly (I︰C). TLR3, NF-κB and IRF3 were detected by real-time PCR. Supernatant fluids from the cultures were analyzed for levels of 6 different pro-inflammatory cytokines, interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-12p70, and TNF-α by flowcytomix measurements. Results With the progress of the disease, the expression of NF-κB was significantly higher in turn, while the level of IRF3 decreased, among three groups. The amounts of IL-10 produced were lower in ACLF than CHB (P = 0.021), but this difference did not achieve statistical significance (P < 0.05) compared to NC. The production of IL-12p70 in MoDC of ACLF was extremely decreased than other two groups (P < 0.001). IL-8 levels were significantly lower in ACLF than in CHB. While, significantly lower amounts were found of IL-8 on MoDCs in CHB than in NC (P < 0.005). IL-1β levels were lower in ACLF than in NC (P = 0.001). Expression of IL-1β was also lower in CHB than in NC (P = 0.007). Conclusions The results suggest impaired TLR3 pathway signaling in MoDC from patients with ACLF, resulting in secretion of selective pro-inflammatory cytokines integral to the inflammatory response that may be critical in the pathogenesis of ACLF.