植物血凝素刺激外周血单个核细胞Th1/Th2细胞因子分泌对阿尔茨海默病的诊断价值

doi:10.3969/j.issn.1673-8640.2018.04.004

摘要/Abstract

摘要：

目的探讨阿尔茨海默病（AD）患者外周血单个核细胞（PBMC）在植物血凝素（PHA）刺激下分泌的细胞因子、血清细胞因子以及血脂水平对疾病的诊断价值。方法收集AD患者血液样本100例以及健康对照者血液样本100例,抽提PBMC,在PHA刺激下培养48 h,采用双抗体夹心酶联免疫吸附试验（ELISA）测定培养液上清中细胞因子白细胞介素（IL）-1α、IL-6、IL-10、肿瘤坏死因子α（TNF-α）以及γ干扰素（IFN-γ）水平;同时检测血清IL-1α（sIL-1α）、血清IL-6（sIL-6）、血清IL-10（sIL-10）、血清TNF-α（sTNF-α）及血清IFN-γ（sIFN-γ）水平和血脂[总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-C）及低密度脂蛋白胆固醇（LDL-C）]水平。6周后AD患者重新采样检测。结果 AD组PBMC分泌的IL-1α、IFN-γ水平明显高于对照组（P<0.05）,而IL-10水平明显低于对照组（P<0.05）;AD组sIL-6及sTNF-α水平明显高于对照组（P<0.05）;AD组血清TC、TG及LDL-C水平明显高于对照组（P<0.05）。AD组6周后重新采样的检测结果与第1次一致。结论在PHA刺激下PBMC的IFN-γ等细胞因子分泌水平、血清细胞因子水平及血脂水平异常可能与AD发病相关,其中PHA刺激的IFN-γ及sIL-6、sTNF-α、TC、TG、LDL-C水平检测对AD有一定的诊断价值。

关键词: 细胞因子, 植物血凝素, 外周血单个核细胞, 阿尔茨海默症

Abstract:

Objective To investigate the roles of cytokines,serum cytokines and blood lipid secreted by phytohaemagglutinin（PHA）-stimulated peripheral blood mononuclear cells（PBMC） for the diagnosis of Alzheimer's disease（AD）. Methods A total of 100 AD patients and 100 healthy subjects were enrolled,and their blood samples were collected. PBMC was extracted. The levels of culture solution cytokine interleukin（IL）-1 alpha,IL-6,IL-10,tumor necrosis factor alpha（TNF-α） and interferon gamma（IFN-γ）,serum cytokines（sIL-1α）,sIL-6,sIL-10,sTNF-α and sIFN-γ and blood lipid [total cholesterol（TC）,triglyceride（TG）,high-density lipoprotein cholesterol（HDL-C） and low-density lipoprotein cholesterol（LDL-C）] levels were determined by enzyme-linked immunosorbent assay（ELISA）. Samples were collected and determined again after 6 weeks. Results The levels of IL-1α and IFN-γ in AD group were higher than those in control group（P<0.05）,while the level of IL-10 in AD group was lower than that in control group（P<0.05）. The levels of sIL-6 and sTNF-α were higher in AD group than those in control group（P<0.05）. The levels of TC,TG and LDL-C in AD group were higher than those in control group（P<0.05）. After 6 weeks,the results of re-sampling were similar those of the first determination. Conclusions Cytokines,serum cytokines and blood lipid secreted by PHA-stimulated PBMC,such as IFN-γ, may be associated with AD. Among them,IFN-γ,sIL-6,sTNF-α,TC,TG and LDL-C may be potential for the diagnosis of AD.

Key words: Cytokine, Phytohaemagglutinin, Peripheral blood mononuclear cell, Alzheimer's disease

中图分类号:

R446.62

丁冰玉, 林萍, 李擎天. 植物血凝素刺激外周血单个核细胞Th1/Th2细胞因子分泌对阿尔茨海默病的诊断价值[J]. 检验医学, 2018, 33(4): 290-294.

DING Bingyu, LIN Ping, LI Qingtian. Th1/Th2 cytokines secreted by phytohaemagglutinin-stimulated peripheral blood mononuclear cells for the diagnosis of Alzheimer's disease[J]. Laboratory Medicine, 2018, 33(4): 290-294.

图/表 5

图1 AD组和对照组PBMC在PHA刺激下分泌的细胞因子水平的比较

图2 AD组和对照组血清细胞因子水平的比较

表1 AD组和对照组血脂水平的比较（mmol/L,x±s）

组别	例数	TC	TG	LDL-C	HDL-C
AD组
AD1	100	6.25±0.77^*	2.20±0.63^*	4.18±0.78^*	1.42±0.41
AD2	100	6.33±1.03^*	2.27±0.41^*	4.32±0.94^*	1.42±0.57
对照组	100	5.27±0.63	1.46±0.69	3.36±0.62	1.47±0.79

图3 评价Th1/Th2、IFN-γ、IL-1α、sIL-6、sTNF-α及血清TC、TG、LDL-C对AD诊断价值的ROC曲线注：（a）评价Th1/Th2（IFN-γ/IL-6比值和IFN-γ/IL-10比值）、IFN-γ、IL-1α对AD诊断价值的ROC曲线; （b）评价IFN-γ、IL-1α、sIL-6以及sTNF-α对AD诊断价值的ROC曲线;（c）评价Th1/Th2（IFN-γ/IL-6比值和IFN-γ/IL-10比值）、IFN-γ及血清TC、TG、LDL-C对AD诊断价值的ROC曲线

表2 ROC曲线评价Th1/Th2、IFN-γ、IL-1α、sIL-6、sTNF-α及血清TC、TG、LDL-C对AD诊断价值的特征性参数

指标	AUC	标准差	P值	95%可信区间
IFN-γ/IL-6比值	0.951	0.023	<0.001	0.905~0.996
IFN-γ/IL-10比值	0.912	0.036	<0.001	0.842~0.982
IFN-γ	0.911	0.037	<0.001	0.838~0.984
IL-1α	0.644	0.082	0.051	0.483~0.805
sIL-6	0.894	0.061	<0.001	0.774~1.000
sTNF-α	0.888	0.064	<0.001	0.761~1.000
TC	0.912	0.042	<0.001	0.829~0.995
TG	0.911	0.044	<0.001	0.825~0.996
LDL-C	0.897	0.049	<0.001	0.801~0.993

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