摘要
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<CHB组<NC组(P<0.005)。IL-1β的分泌水平在ACLF组与NC组(P=0.001)、CHB组与NC组(P=0.007)间差异均有显著统计学意义。结论 ACLF患者单核细胞来源的树突状细胞TLR3信号转导通路受损,促炎性细胞因子分泌异常,可能是肝细胞免疫功能严重受损的原因之一。
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.
出处
《中国肝脏病杂志（电子版）》
CAS
2012年第1期5-9,共5页
Chinese Journal of Liver Diseases：Electronic Version
基金
国家自然科学基金(81101240)
国家十一五重大科技专项(2008ZX10002-004)
