单核细胞亚群在视神经脊髓炎中的变化及糖皮质激素对其的影响

发布时间：2018-03-21 06:48

本文选题：视神经脊髓炎　切入点：糖皮质激素　出处：《中南大学》2014年硕士论文　论文类型：学位论文

【摘要】：背景：单核细胞作为重要的固有免疫细胞,具有明显的异质性,根据细胞表面CD16分子表达水平,分为经典型亚群(CD14++CD16-)、中间型亚群(CD14++CD16+)和非经典型亚群(CD14+CD16++),促炎细胞因子IL-1β和TNF-α为非经典型亚群分泌的主要细胞因子。CD16+亚群较CD16-亚群具有更强的促炎作用,其在多种炎性疾病,尤其是自身免疫性疾病中出现明显扩增,分泌的促炎细胞因子IL-1β和TNF-α增加,因而认为其在自身免疫性疾病的发病过程中具有重要作用。视神经脊髓炎(NMO)是中枢神经系统(CNS)自身免疫性疾病,活化的单核细胞是最先到达CNS炎症部位的免疫细胞之一,促炎细胞因子IL-1β和TNF-α参与了NMO发生的病理过程。但目前单核细胞三个亚群在NMO中的变化情况及常规的NMO治疗药物糖皮质激素对单核细胞亚群的影响尚有待阐明。目的：研究单核细胞三亚群(经典型、中间型和非经典型)在NMO患者中的变化情况,探讨其在NMO发病中的可能作用机制。进一步观察糖皮质激素冲击治疗对单核细胞亚群分布的影响,为应用针对单核细胞及其细胞因子的治疗方法治疗NMO提供理论依据。方法：本研究纳入20例急性期NMO患者,以15例健康体检者及10例非炎症性中枢神经系统疾病(NIND)患者为对照；NMO患者分别于糖皮质激素冲击治疗前及冲击治疗结束时行EDSS评分。通过对外周血单个核细胞进行CD14及CD16鼠抗人单克隆抗体染色,流式细胞仪检测,根据单核细胞上CD14和CD16的表达情况,将单核细胞分为三个亚群：CD14++CD16-(经典型)、CD14++CD16+(中间型)和CD14+CD16++(非经典型),从而获得单核细胞三个亚群所占百分比数据。实时荧光定量PCR检测单核细胞IL-1β、TNF-αmRNA表达及ELISA方法检测血浆及脑脊液中IL-1β、TNF-α含量。分析比较各组之间在上述指标之间的差异性及相关性。使用SPSS17.0软件包进行统计学处理。结果：NMO患者较正常对照组单核细胞非经典型亚群及中间型亚群明显扩增,而经典型亚群则明显减少；糖皮质激素治疗后NMO患者单核细胞非经典型亚群出现显著减少,中间型亚群比例无明显变化,而单核细胞经典型亚群则明显增加；NMO患者治疗后EDSS评分较治疗前EDSS评分有显著改善：单核细胞非经典型及经典型亚群治疗前后的变化与治疗前后EDSS评分改善呈正相关,而中间型亚群变化则与EDSS改善无相关性；治疗前NMO患者,单核细胞IL-1β、TNF-α mRNA相对表达及血浆、脑脊液IL-1β、TNF-α水平升高,经糖皮质激素冲击治疗后均较治疗前下降；治疗前NMO患者IL-1β、TNF-α mRNA相对表达与非经典型单核细胞亚群比例呈正相关,与中间型单核细胞亚群及经典型单核细胞亚群比例无相关性。NMO患者治疗前后血浆IL-1β、TNF-α变化与EDSS评分改善呈正相关。结论： 1、NMO患者CD16+单核细胞比例增高,CD16-单核细胞比例减少,存在CD16+/CD16-单核细胞比例失衡； 2、急性期NMO患者非经典型单核细胞亚群增高可能通过增加IL-1β、TNF-α的表达与分泌,参与NMO疾病的发生； 3、糖皮质激素冲击治疗可通过抑制单核细胞非经典型亚群的扩增及IL-1β、 TNF-α的表达分泌,纠正CD16+/CD16-单核细胞失衡,发挥对NMO的治疗作用。
[Abstract]:Background: mononuclear cells as important innate immune cells, with obvious heterogeneity, based on cell surface expression of CD16 molecules, divided into classical subsets (CD14++CD16-), intermediate subgroup (CD14++CD16+) and non classical subsets (CD14+CD16++), proinflammatory cytokines IL-1 and TNF- alpha - beta the classical subsets cytokine secretion of the major subgroup.CD16+ than CD16- subgroup has stronger proinflammatory effects, which in a variety of inflammatory diseases, especially autoimmune diseases with obvious amplification, the secretion of proinflammatory cytokines and TNF- alpha IL-1 beta increased, so that it plays an important role in the pathogenesis of their own autoimmune diseases in neuromyelitis optica (NMO) is a central nervous system autoimmune disease (CNS), monocyte activation of immune cells is one of the CNS sites of inflammation at first, proinflammatory cytokines IL-1 and TNF- alpha beta in NMO The pathological process of birth. But at present, the change of three subsets of monocytes in NMO and the effect of conventional NMO therapy glucocorticoids on monocyte subsets need to be elucidated.
Objective: To study the mononuclear cells of Sanya group (classic type, intermediate type and non classical) changes in NMO patients and to explore its possible role in the pathogenesis of NMO. The mechanism to further observe the impact of glucocorticoid treatment on monocyte subsets, and provide a theoretical basis for the treatment methods for the single nuclear cells and the treatment of NMO.
Methods: the study included 20 cases of acute NMO patients, 15 healthy subjects and 10 patients with non inflammatory diseases of the central nervous system (NIND) were selected as control; NMO patients were treated with corticosteroid treatment before and after treatment by EDSS scores. The impact of CD14 and CD16 monoclonal antibody staining by peripheral blood mononuclear cells, flow cytometry, according to the expression of mononuclear cells of CD14 and CD16, the mononuclear cells were divided into three subgroups: CD14++CD16- (classical), CD14++CD16+ (intermediate) and CD14+CD16++ (non classical), so as to obtain mononuclear cells of three subgroups the percentage of data. Real time fluorescence quantitative PCR detection of monocyte IL-1 beta, plasma and cerebrospinal fluid detection of alpha mRNA expression and ELISA method in TNF- IL-1 beta, TNF- alpha content. Analysis and comparison between the differences in the above indexes and the correlation between the use of SPSS17. .0 software packages are statistically processed.
Results: NMO patients compared with normal control group, mononuclear cells of non classical subsets and intermediate subsets significantly amplified, and classical subsets decreased; after glucocorticoid treatment of monocytes in patients with NMO non classical subsets significantly reduced, intermediate subgroup had no obvious change, while the single core classic cell subsets increased significantly; NMO patients EDSS score before treatment EDSS score improved significantly: mononuclear cells and non changes before and after the treatment of classic and classic subsets of treatment EDSS score improvement was positively correlated, and changes of subsets and EDSS to improve the relevance of NMO before treatment; patients, monocyte IL-1 beta, TNF- alpha and mRNA expression in plasma, CSF IL-1 beta, elevated TNF- levels, the impact of glucocorticoid treatment were decreased than before treatment; before treatment in patients with NMO IL-1 beta, TNF- alpha and mRNA expression The proportion of classical monocyte subsets is positively correlated, and has no correlation with the proportion of intermediate monocyte subsets and classical monocyte subsets. The changes of plasma IL-1 beta and TNF- alpha in.NMO patients are positively correlated with the improvement of EDSS score.
Conclusion:
1, the proportion of CD16+ mononuclear cells in NMO patients was higher, and the proportion of CD16- monocytes decreased, and the proportion of CD16+/CD16- mononuclear cells was unbalance.
2, the increase of non classical monocyte subsets in acute NMO patients may be involved in the occurrence of NMO disease by increasing the expression and secretion of IL-1 beta, TNF- alpha.
3, glucocorticoid pulse therapy can suppress the imbalance of CD16+/CD16- monocyte and play a role in the treatment of NMO by inhibiting the amplification of monocyte non classical subpopulations and the expression and secretion of IL-1 and TNF-.

【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R744.52

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