HVHF治療對老年膿毒癥患者外周血單核細胞miRNA-155表達的影響
發(fā)布時間:2019-07-21 12:37
【摘要】:目的探討高通量連續(xù)性血液濾過技術(shù)(HVHF)治療對老年膿毒癥患者外周血單核細胞(PBMC)miRNA-155表達的影響。方法老年嚴重膿毒癥患者60例,均接受對癥支持治療,根據(jù)是否接受HVHF治療分為HVHF組(29例)和膿毒癥組(31例)。分別于治療前,治療第4、8、12、24、48小時采集肘靜脈血,分離PBMC,熒光定量PCR檢測miRNA-155表達量。在治療24 h后分離兩組PBMC進行體外培養(yǎng),用脂多糖刺激4、8、12、24、48 h時,檢測miRNA-155表達量,并用酶聯(lián)免疫吸附(ELISA)法檢測細胞培養(yǎng)液中白細胞介素(IL)-6、IL-10和腫瘤壞死因子(TNF)-α水平。結(jié)果兩組治療過程中,隨著時間延長兩組miRNA-155表達量均明顯降低(P0.05);其中HVHF組下降幅度明顯大于膿毒癥組,且各時間點miRNA-155表達量均明顯低于膿毒癥組(均P0.05)。脂多糖刺激后HVHF組PBMC miRNA-155表達量明顯高于孵育前,孵育12 h時miRNA-155表達量達到峰值;膿毒癥組PBMC miRNA-155表達量上升幅度明顯低于HVHF組,孵育8 h時達到峰值;孵育前和孵育4、8 h時膿毒癥組miRNA-155表達量均明顯高于同期的HVHF組(P0.05)。脂多糖刺激后HVHF組PBMC IL-6、TNF-α分泌量持續(xù)增加,12 h達到峰值。膿毒癥組PBMC IL-6、TNF-α分泌量在8 h達到峰值。在PBMC孵育的各時間點,HVHF組IL-6、TNF-α分泌量均明顯高于膿毒癥組(P0.05)。兩組PBMC IL-10分泌量在8、12、24、48 h均為處在高水平,且各時間點兩組之間差異均無統(tǒng)計學(xué)意義(P0.05)。結(jié)論 HVHF治療可通過間接下調(diào)PBMC miRNA-155表達水平,減輕其對炎性因子釋放的抑制作用,增強對外界再刺激的反應(yīng)能力,改善老年嚴重膿毒癥患者的治療預(yù)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)-偽 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- 偽 in HVHF group continued to increase and reached the peak at 12 h. The secretion of PBMC IL-6,TNF- 偽 in sepsis group reached the peak at 8 h. At each time point of PBMC incubation, the secretion of IL-6,TNF- 偽 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.
【作者單位】: 首都醫(yī)科大學(xué)附屬北京世紀壇醫(yī)院重癥醫(yī)學(xué)科;
【分類號】:R459.7
[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)-偽 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- 偽 in HVHF group continued to increase and reached the peak at 12 h. The secretion of PBMC IL-6,TNF- 偽 in sepsis group reached the peak at 8 h. At each time point of PBMC incubation, the secretion of IL-6,TNF- 偽 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.
【作者單位】: 首都醫(yī)科大學(xué)附屬北京世紀壇醫(yī)院重癥醫(yī)學(xué)科;
【分類號】:R459.7
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