HVHF治疗对老年脓毒症患者外周血单核细胞miRNA-155表达的影响

发布时间：2019-07-21 12:37

【摘要】：目的探讨高通量连续性血液滤过技术(HVHF)治疗对老年脓毒症患者外周血单核细胞(PBMC)miRNA-155表达的影响。方法老年严重脓毒症患者60例,均接受对症支持治疗,根据是否接受HVHF治疗分为HVHF组(29例)和脓毒症组(31例)。分别于治疗前,治疗第4、8、12、24、48小时采集肘静脉血,分离PBMC,荧光定量PCR检测miRNA-155表达量。在治疗24 h后分离两组PBMC进行体外培养,用脂多糖刺激4、8、12、24、48 h时,检测miRNA-155表达量,并用酶联免疫吸附(ELISA)法检测细胞培养液中白细胞介素(IL)-6、IL-10和肿瘤坏死因子(TNF)-α水平。结果两组治疗过程中,随着时间延长两组miRNA-155表达量均明显降低(P0.05);其中HVHF组下降幅度明显大于脓毒症组,且各时间点miRNA-155表达量均明显低于脓毒症组(均P0.05)。脂多糖刺激后HVHF组PBMC miRNA-155表达量明显高于孵育前,孵育12 h时miRNA-155表达量达到峰值;脓毒症组PBMC miRNA-155表达量上升幅度明显低于HVHF组,孵育8 h时达到峰值;孵育前和孵育4、8 h时脓毒症组miRNA-155表达量均明显高于同期的HVHF组(P0.05)。脂多糖刺激后HVHF组PBMC IL-6、TNF-α分泌量持续增加,12 h达到峰值。脓毒症组PBMC IL-6、TNF-α分泌量在8 h达到峰值。在PBMC孵育的各时间点,HVHF组IL-6、TNF-α分泌量均明显高于脓毒症组(P0.05)。两组PBMC IL-10分泌量在8、12、24、48 h均为处在高水平,且各时间点两组之间差异均无统计学意义(P0.05)。结论 HVHF治疗可通过间接下调PBMC miRNA-155表达水平,减轻其对炎性因子释放的抑制作用,增强对外界再刺激的反应能力,改善老年严重脓毒症患者的治疗预后。
[Abstract]:Objective to investigate the effect of high throughput continuous hemofiltration (HVHF) on the expression of (PBMC) miRNA-155 in peripheral blood monocytes of elderly patients with sepsis. Methods 60 elderly patients with severe sepsis were treated with symptomatic support. According to whether they were treated with HVHF, they were divided into HVHF group (29 cases) and sepsis group (31 cases). The blood samples of elbow vein were collected at 48 hours before treatment, and the expression of miRNA-155 was detected by PBMC, fluorescence quantitative PCR. After 24 hours of treatment, the two groups of PBMC were isolated and cultured in vitro and stimulated with lipopolysaccharide for 4, 8, 12, 24 and 48 h. The expression of miRNA-155 was detected by enzyme-linked immunosorbent assay (ELISA). The levels of IL-6, IL-10 and (TNF)-伪 in the cell culture medium were detected by enzyme-linked immunosorbent assay (Elisa). Results during the treatment process, the expression of miRNA-155 in both groups decreased significantly with the prolongation of time (P 0.05), and the expression of miRNA-155 in HVHF group was significantly higher than that in sepsis group, and the expression of miRNA-155 in both groups was significantly lower than that in sepsis group at each time point (P 0.05). After lipopolysaccharide stimulation, the expression of PBMC miRNA-155 in HVHF group was significantly higher than that before incubation, and the expression of miRNA-155 in sepsis group reached the peak at 12 h, while the PBMC miRNA-155 expression in sepsis group was significantly lower than that in HVHF group and reached the peak at 8 h after incubation, and the expression of miRNA-155 in sepsis group was significantly higher than that in HVHF group before incubation and 4 h after incubation (P 0.05). After lipopolysaccharide stimulation, the secretion of PBMC IL-6,TNF- 伪 in HVHF group continued to increase and reached the peak at 12 h. The secretion of PBMC IL-6,TNF- 伪 in sepsis group reached the peak at 8 h. At each time point of PBMC incubation, the secretion of IL-6,TNF- 伪 in HVHF group was significantly higher than that in sepsis group (P 0.05). The secretion of PBMC IL-10 in the two groups was at a high level at 8, 12, 24 and 48 h, and there was no significant difference between the two groups at each time point (P 0.05). Conclusion HVHF treatment can indirectly down-regulate the expression of PBMC miRNA-155, reduce its inhibitory effect on the release of inflammatory factors, enhance the ability to respond to external restimulation, and improve the prognosis of elderly patients with severe sepsis.
【作者单位】：首都医科大学附属北京世纪坛医院重症医学科;
【分类号】：R459.7

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