不同置换量血液滤过对脓毒症患者Toll样受体表达的影响  

Effects of hemodiafiltration with different amount of substitute fluid on toll-like receptors in sepitc patients

作　　者：茅尧生[1] 吕铁[1] 郑建鹏[1] 

机构地区：[1]绍兴市人民医院危重病科,浙江省绍兴312000

出　　处：《中华急诊医学杂志》2009年第10期1047-1051,共5页Chinese Journal of Emergency Medicine

基　　金：浙江省医药卫生科学研究基金（2007A186）

摘　　要：目的前瞻性的研究不同置换量血液滤过对严重脓毒症患者外周血单核细胞Toll样受体表达及其内在功能的影响。方法对2006年3月至2009年2月入住绍兴市人民医院ICU符合纳入标准的严重脓毒症患者30例，在脓毒症集束化治疗策略的基础上联合血液滤过治疗，并随机分为高通量血液滤过组（HVHF）（置换量4L／h，15例）与常规通量血液滤过组（置换量2L／h，15例）。①血液滤过治疗0h，6h，12h，24h，48h各时相点，分离外周血单核细胞，采用半定量RT-PCR法和流式细胞仪榆测单核细胞表面Toll样受体-4（TLR4）和Toll样受体-2（TLR4）mRNA和蛋白表达变化。②血液滤过治疗24h后分离患者外周血单核细胞于休外培养，以内毒素（LPS）刺激，在6h，12h，24h，48h各时相点检测单核细胞表面TLR4和TLR2受体mRNA和蛋白的变化，及ELISA法检测单核细胞培养液肿瘤坏死因子-α（TNF-α），白介素-6（IL-6），白介素-8（IL-8）细胞因子水平。统计学方法：对计量资料采用均数±标准差描述，采用t检验或Oneway ANOVA分析。结果高通量血液滤过较常规通量可显著下调单核细胞TLR2和TLR4 mRNA及蛋白表达水平（P〈0．05）。不同置换量血滤治疗处理后孵育的单核细胞，其TNF-α，IL-6，IL-8分泌水平各时相点呈现：常规通量组〉高通量组〉正常对照，P〈0．05；TLR4和TLR2 mRNA各时相点表达上调水平。呈现：正常对照〉高通量组〉常规通量组，P〈0．05；其TLR2和TLR4蛋白表达亦呈现mRNA相同趋势，差异无统计学意义。结论高通量血液滤过能改善脓毒症患者单核细胞功能，使部分细胞免疫功能得到恢复，对重建机体免疫内稳状态起一定作用。Objective To prospectively evaluate the effect of hemodiafihration with different amount of substitute fluid on expression and function of Toll-like receptor on peripheral blood monocyte in severe septic patients. Method Thirty severe septic patients accepted in ICU of Shaoxing People＇s Hospital from March 2006 to February 2009 were qualified for bundle strategies therapy plus hemodiafiltration, and they were randomly derided into two groups with fifteen persons in each one： high volume hemofihration group （HVHF4 L/hr） and continuous veno-venous hemofihration group （CWH 2 L/hr）. ① The peripheral blood monocytes were isolated at specific points of hemofihration treatment（0 h,6 h, 12 h,24 h,48 h） ; Toll-like reeepto-2（TLR2）, reeeptor-4（TLR4） mRNA and protein on surface were analyzed by scmiquantitative RT-PCR and flow cytometry. ② The peripheral blood monoeytes were isolated 24 h after hemofiltration and underwent incubation in vitro with stimulation of （ lipopolysaecharide）LPS;the expression of TLR2/TLR4 mRNA and protein were determined at 6 h, 12 h,24 h and 48 h point of incubation repectively; and the levels of TNF-α, IL-6, IL-8 in the substrate fluid were also tested by ELISA. All the data were presented as x ± s and analyzed by t -test or one way ANOVA. Results The expression of TLR2/ TLR4 mRNA and protein more decreased in HVHF group （ P 〈 0.05）. The levels of TNF-α, IL-6, IL-8 secreted by the incubated monocytes in each specific time point varied in terms of the amount of substitute fluid： CWH 〉 HVHF 〉 controlled group （ P 〈 0.05）. Both the expression of TLR2/TLR4 mRNA and protein on surface up-regulated at each time point： controlled group 〉 HVHF 〉 CVVH （mRNA P 〈 0.05; protein P 〉 0.05）. Conclusions HVHF cotdd improve the function of monocytes to partially recover the celluar-immunity and may play a role in restoration of immunal homeostasis in septic patients.

关 键 词：脓毒症 血液净化 高通量血液滤过 Toll样受体-2 TOLL样受体-4 肿瘤坏死因子-α 白介素-6 白介素-8 单核细胞培养 半定量RT-PCR 

分 类 号：R459.7[医药卫生—急诊医学]