脓毒症患者外周血DC成熟与分泌的变化及OXPAPC的影响

发布时间：2018-11-27 08:08

【摘要】：背景：脓毒症患者树突状细胞(DC)的成熟度与分泌功能受到抑制,而DC活性的恢复有助于脓毒症的好转,持续LPS刺激是体外模拟脓毒症感染的常用且有效方式,且持续LPS对DC的效应亦为抑制。因此探讨能减轻DC成熟与分泌的抑制效应的因素,对脓毒症的发展与调控有重要意义。在此基础上,寻找减轻持续LPS对DC成熟与分泌功能抑制作用的药物,能为脓毒症的治疗提供理论依据。目的：观察脓毒症患者外周血DC成熟度与分泌功能的变化及不同浓度OXPAPC对其的影响,初步探讨OXPAPC对脓毒症作用的机制。方法：①选择健康志愿者和脓毒症患者无菌条件下获得外周血中单个核细胞(包括淋巴细胞和单核细胞)。体外使用GM-CSF、IL-4和TNF-α诱导其定向DC分化成熟。在倒置显微镜和透射电镜下观察细胞形态,流式细胞仪检测CDla、HLA-DR和CD86的表达水平差异,ELISA法检测培养上清液IL-12p70的变化。 ②采用持续性LPS刺激单个核细胞模拟体内脓毒症中DC反应。无菌条件下分离健康志愿者外周血单个核细胞,使用GM-CSF、IL-4和TNF-α诱导单个核细胞定向DC分化成熟,加入持续性LPS和不同浓度OXPAPC进行干预。在倒置显微镜和透射电镜下观察细胞形态,流式细胞仪检测CDla、HLA-DR和CD86的表达水平差异,ELISA法检测培养上清液IL-12p70的变化。结果：①与健康志愿者相比,脓毒症患者组HLA-DR、CD86的表达水平及IL-12p70的分泌水平均较低,均P0.05。 ②持续LPS刺激干预组HLA-DR、CD86的表达水平及IL-12p70的水平均较对照组降低,均P0.05、OXPAPC成浓度依赖性地升高持续LPS刺激下HLA-DR、CD86和IL-12p70的水平,与持续LPS刺激组比较,差异均有统计学意义(均P0.05)。结论：1.脓毒症患者外周血DC的成熟度与分泌功能是受抑制的；2.OXPAPC成浓度依赖性减轻持续LPS刺激对DC成熟度和分泌功能的抑制效应。
[Abstract]:Background: the maturity and secretory function of dendritic cells (DC) in septic patients are inhibited, and the recovery of DC activity contributes to the improvement of sepsis. Continuous LPS stimulation is a common and effective way to simulate sepsis infection in vitro. The effect of continuous LPS on DC was also inhibited. Therefore, it is important for the development and regulation of sepsis to explore the factors that can reduce the inhibitory effect of DC maturation and secretion. On this basis, to find a drug to reduce the inhibitory effect of LPS on DC maturation and secretory function, can provide theoretical basis for the treatment of sepsis. Aim: to observe the changes of DC maturity and secretory function in peripheral blood of septic patients and the effect of different concentrations of OXPAPC on it, and to explore the mechanism of OXPAPC on sepsis. Methods: 1 Mononuclear cells (including lymphocytes and monocytes) in peripheral blood were obtained from healthy volunteers and septic patients. GM-CSF,IL-4 and TNF- 伪 were used to induce DC differentiation and maturation in vitro. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. 2 continuous LPS was used to stimulate mononuclear cells to mimic the DC response in sepsis in vivo. The peripheral blood mononuclear cells (PBMC) of healthy volunteers were isolated under aseptic conditions. GM-CSF,IL-4 and TNF- 伪 were used to induce the differentiation and maturation of mononuclear cells (MNCs), and continuous LPS and different concentrations of OXPAPC were added to the mononuclear cells. The cell morphology was observed under inverted microscope and transmission electron microscope. The expression levels of CDla,HLA-DR and CD86 were detected by flow cytometry and the changes of IL-12p70 in culture supernatant were detected by ELISA method. Results: 1 the expression of HLA-DR,CD86 and the secretion of IL-12p70 in sepsis patients were lower than those in healthy volunteers (P 0.05). 2 the expression of HLA-DR,CD86 and the level of IL-12p70 in the intervention group of continuous LPS stimulation were lower than those in the control group, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner by P0.05OXPAPC, and the levels of HLA-DR,CD86 and IL-12p70 were increased in a concentration-dependent manner. Compared with the continuous LPS stimulation group, the difference was statistically significant (P0.05). Conclusion: 1. The maturity and secretory function of DC in peripheral blood of septic patients were inhibited, and the inhibitory effect of continuous LPS stimulation on DC maturity and secretory function was alleviated in a concentration-dependent manner by 2.OXPAPC.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7

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