NETs在癫痫患者癫痫发作后血浆、脑脊液中表达变化研究

发布时间：2018-11-08 09:22

【摘要】：目的白细胞向脑内浸润引起炎症反应在癫痫的发生和发展过程中起着极其重要的作用。然而,癫痫发作后白细胞引起神经元损伤的具体机制尚不清楚。中性粒细胞胞外诱捕网(Neutrophil extracellular traps,NETs)是由中性粒细胞释放到胞外的包含有DNA、组蛋白、髓过氧化物酶等抗菌蛋白的纤维网状结构,在感染性疾病中扮演着重要的角色,并能加重神经元损害。本研究观察全面强直阵挛性发作(Generalized tonic-clonic seizures,GTCS)癫痫患者发作后,脑脊液(cerebrospinal fluid,CSF)和血浆(Plasma)中NETs(dsDNA、MPO)含量及血浆中S100B水平,并分析NETs与S100B的相关性,探讨NETs与血脑屏障通透性关系,为探索癫痫治疗的新靶点提供理论依据。方法选取2015年11月至2016年12月入住西安医学院第一附属医院神经内科的GTCS癫痫患者31例及紧张性头痛患者27例,并于发病24h内进行血浆、脑脊液标本的采集。采用PicoGreen dsDNA Quantitation kits和酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)分别测定癫痫组与对照组血浆及脑脊液中NETs(dsDNA、MPO)和S100B含量,并分析二者之间的相关性。符合正态分布的计量资料用x±s表示,不符合正态分布的计量资料用M(LQ,UQ)表示,两组独立样本比较采用T检验(符合正态分布、方差齐的计量资料)和Mann-Whitney U检验(不符合正态分布、方差不齐的计量资料),两组相关数据间比较采用Wilcoxon带符号秩检验;性别构成比等分类变量资料采用?2检验,数据相关性分析采用Spearman秩相关系数检验。所有统计学分析中,(49)(27)0.05认为差异有统计学意义。结果癫痫组血浆、脑脊液中dsDNA的含量明显高于对照组((49)(27)0.001;(49)(27)0.05),血浆、脑脊液中MPO的含量高于对照组((49)(27)0.05);癫痫组S100B水平明显高于对照组((49)(27)0.05);癫痫组脑脊液/血浆中dsDNA和MPO水平与血浆S100B含量均不具有相关性((49)0.05)。结论GTCS癫痫患者发作后血浆和脑脊液中NETs(dsDNA、MPO)浓度均升高,说明NETs(dsDNA、MPO)可能参与了癫痫的发生与发展。GTCS癫痫患者血浆中S100B水平较对照组明显升高,证实了癫痫发作与血脑屏障破坏有关。GTCS癫痫患者脑脊液、血浆中NETs(dsDNA、MPO)含量与血浆S100B无相关性,提示NETs的形成与GTCS癫痫患者发作后血脑屏障破坏可能无关。
[Abstract]:Objective inflammation induced by leukocyte infiltration into the brain plays an important role in the occurrence and development of epilepsy. However, the mechanism of leukocyte-induced neuronal damage after seizure is unclear. Neutrophil extracellular entrapment (Neutrophil extracellular traps,NETs) is a fibrous reticular structure containing DNA, histone, myeloperoxidase and other antibacterial proteins released from neutrophil to extracellular, which plays an important role in infectious diseases. And can aggravate neuronal damage. In this study, the contents of NETs (dsDNA,MPO) in cerebrospinal fluid (cerebrospinal fluid,CSF) and plasma (Plasma) and the level of S100B in plasma were observed in patients with generalized tonic-clonic seizure (Generalized tonic-clonic seizures,GTCS) after seizure, and the correlation between NETs and S100B was analyzed. To explore the relationship between NETs and blood-brain barrier permeability, and to provide theoretical basis for exploring new targets of epilepsy therapy. Methods from November 2015 to December 2016, 31 patients with GTCS epilepsy and 27 patients with tension headache were enrolled in the Department of Neurology, first affiliated Hospital of Xi'an Medical College. The samples of plasma and cerebrospinal fluid (CSF) were collected within 24 hours after onset. The contents of NETs (dsDNA,MPO) and S100B in plasma and cerebrospinal fluid of epileptic group and control group were determined by PicoGreen dsDNA Quantitation kits and enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay,ELISA), and the correlation between them was analyzed. The metrological data in accordance with normal distribution are expressed in x 卤s, and those that do not conform to normal distribution are expressed in M (LQ,UQ). The two groups of independent samples are compared by T test (consistent with normal distribution). Mann-Whitney U test (not in accordance with the normal distribution, the measurement data with uneven variance) and the Wilcoxon signed rank test were used in the comparison between the two groups of related data. The data of sex composition ratio and other classified variables were tested by? 2 test, and Spearman rank correlation coefficient test was used for data correlation analysis. In all statistical analysis, (49) (27) 0.05 thought the difference was statistically significant. Results the levels of dsDNA in plasma and cerebrospinal fluid in epilepsy group were significantly higher than those in control group (49) (27) and (49) (27). The contents of MPO in plasma and cerebrospinal fluid were higher than those in control group (49) (27). The level of S100B in epileptic group was significantly higher than that in control group (49) (27). There was no correlation between the levels of dsDNA and MPO in cerebrospinal fluid / plasma and the content of S100B in epileptic group (49) 0.05). Conclusion the concentration of NETs (dsDNA,MPO) in plasma and cerebrospinal fluid (CSF) of GTCS epileptic patients increased after seizure, indicating that NETs (dsDNA,MPO) may be involved in the occurrence and development of epilepsy. The level of S100B in GTCS epileptic patients was significantly higher than that in control group. There was no correlation between the content of NETs (dsDNA,MPO) in plasma and S100B in patients with GTCS epilepsy, suggesting that the formation of NETs may not be related to the breakdown of blood-brain barrier in GTCS epileptic patients.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.1

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