摘要
目的前瞻性观察高通量连续性血液滤过（HVHF）治疗对严重脓毒症患者免疫功能及其外周血单个核细胞中miRNA-146a表达的影响。方法选择严重脓毒症患者30例，分为HVHF组及对照组。①在两组治疗过程的0、6、12、24、48h时相点，分离外周血单个核细胞，提取总RNA，荧光定量PCR法检测miRNA-146a表达量的变化；②两组患者治疗24h后分离出外周血单个核细胞于体外培养，以LPS刺激，在4、8、12、24、48h时相点检测单个核细胞中miRNA-146a表达量的变化；ELISA法检测单个核细胞培养液中TNF-仅、IL-6、IL-10水平。结果①两组患者miRNA-146a表达量较治疗前均有所下调，但HVHF组较对照组下调明显（P〈0．05）；②HVHF组孵育的细胞在LPS刺激下各个时相TNF-OL、IL-6的分泌水平均明显高于对照组（P〈0．05）；HVHF组TNF—ol、IL-6在12h分泌达高峰，对照组在8h即达高峰；两组细胞IL-10分泌水平均持续升高，两组间各时相点IL-10分泌水平比较差异无统计学意义（P〉0．05）；③HVHF组孵育的细胞在LPS刺激下miRNA-146a表达量较孵育前有所升高，在12h升高最明显（P〈0．05），对照组在4、8h时miRNA-146a表达都处于高水平状态，明显高于同时相的HVHF组（P〈0．05）。结论HVHF通过大量清除脓毒症患者血循环中的炎症因子及促炎介质，从而下调部分miRNA-146a的表达水平，减轻严重脓毒症患者外周血单个核细胞的免疫抑制状态，使部分免疫功能得到恢复，可能是HVHF参与机体免疫内稳状态重建的作用机制之-。
Objective To prospectively evaluate the effect of High volume hemofltration （HVHF） on immune function and expression of miRNA - 146a in the peripheral blood mononuelear cells in severe septic patients. Methods Select 30 cases of severe sepsis patients,divided into HVHF and control groups. ①During the treatment 0 h, 6 h, 12 h, 24 h, 48 h point in two groups, Separate mononuclear cell from peripheral blood and extracted total RNA with miRNeasy Mini Kit and take real - time PCR to detect the miRNA - 146a expression. ②The peripheral blood mononuclear cell were isolated 24 h after treatment, Stimulation of LPS,the expression of miRNA - 146a were determined at 4 h,8 h, 12 h,24 h and 48 h point of incubation repectively;and the levels of TNF - α, IL -6, IL - 10 in the substrate fluid were also tested by ELISA. Results ①Two treatment groups of miR - 146a expression are higher than the pre - treatment, the HVHF group was lower significantly than the control group （ P 〈 0.05） ;②The mononuclear cells of HVHF group stimulated by LPS, in various phases, the secretion ofTNF - α, IL - 6 was significantly higher than the control group （ P 〈 0.05 ）. HVHF group of TNF -α, IL-6 secretion levels at 12 h secretion reached a peak but control group reached a peak at 8 h. Two groups of IL - 10 average secretion levels continues to rise and no significant difference between the two groups at each time points （ P 〉 0.05 ） ; ③HVHF group of miRNA - 146a expression was higher than before incubation, significantly at 12 h （P 〈 O. 05 ）, in the point of 4 h, 8 h after stimulation by LPS, the expression of miRNA -146a are higher significantly than the HVHF group（ P 〈 O. 05 ）. Conclusion HVHF can effectively remove the excessive release of inflammatory cytokines in the peripheral blood of patients with sepsis, and by down - regulating peripheral blood mononuclear cells of miR - 146a expression levels, improved the reactivity of stimulation from the external. Made the immune function of some cells to recovery, involved in the reconstruction of the immune steady state.
出处
《中国急救医学》
CAS
CSCD
北大核心
2012年第6期497-501,共5页
Chinese Journal of Critical Care Medicine
基金
绍兴市社会发展重点科研项目（2011A23021）
