临床研究
ENGLISH ABSTRACT
不同类型过敏性结膜炎患者眼表组织中白细胞介素-4、5和13的表达
姚娟
李冰
郑晓汾
作者及单位信息
·
DOI: 10.3760/cma.j.issn.2095-0160.2015.10.013
Expressions of interleukin-4, 5 and 13 in ocular surface with different types of allergic conjunctivitis
Yao Juan
Li Bing
Zheng Xiaofen
Authors Info & Affiliations
Yao Juan
Shanxi Eye Hospital, Taiyuan 030002, China
Li Bing
Zheng Xiaofen
·
DOI: 10.3760/cma.j.issn.2095-0160.2015.10.013
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摘要

背景过敏性结膜炎的发病机制尚未完全明确。研究表明,白细胞介素-4(IL-4)、IL-5和IL-13在过敏性结膜炎中发挥重要作用,但这些因子对不同类型的过敏性结膜炎患者的作用是否一致尚有待证明。

目的探讨IL-4、IL-5和IL-13在不同类型过敏性结膜炎患者眼表组织中的表达及其在不同类型过敏性结膜炎发病中的作用。

方法采用前瞻性队列研究设计。纳入2013年4月至2014年9月在山西省眼科医院就诊的春季角结膜炎(VKC)患者20例40眼、季节性过敏性结膜炎(SAC)患者20例40眼和常年性过敏性结膜炎(PAC)患者20例40眼,并纳入同期健康志愿者20人40眼作为正常对照。用消毒刮刀在上睑结膜刮取结膜上皮细胞,分别采用免疫组织化学法、实时荧光定量PCR法检测结膜上皮细胞中IL-4、IL-5和IL-13蛋白及其mRNA的表达;采用1 μl毛细玻璃管于受试者下睑颞侧睑缘处吸取泪液4 μl,Luminex液相芯片测定受试者泪液中IL-4、IL-5和IL-13的表达。

结果免疫组织化学法染色表明,VKC组、SAC组和PAC组患者结膜上皮细胞的细胞质中均可见到IL-4、IL-5和IL-13的阳性表达,而在正常对照组受试者结膜上皮细胞中未发现IL-4、IL-5和IL-13的表达。以正常对照组IL-4、IL-5和IL-13 mRNA的表达量为对照,VKC组、SAC组和PAC组中IL-4 mRNA的相对表达量分别为4.11±1.24、2.71±0.71和2.00±0.80,IL-5 mRNA的相对表达量分别为4.02±0.43、2.07±0.45和1.47±0.50,IL-13 mRNA的相对表达量分别为6.44±0.66、4.35±1.26和2.39±0.86,4个组间IL-4、IL-5和IL-13 mRNA相对表达量的总体比较差异均有统计学意义( F=51.32、220.18、162.49,均 P<0.01);其中VKC组IL-4、IL-5和IL-13 mRNA的相对表达量明显高于SAC组和PAC组,SAC组IL-4、IL-5、IL-13 mRNA的相对表达量高于PAC组,差异均有统计学意义(均 P<0.05)。正常对照组受试者泪液中未检测到IL-4、IL-5和IL-13表达,VKC组泪液中IL-4、IL-5和IL-13蛋白质量浓度分别为(14.06±3.50)、(10.88±1.82)和(34.28±8.42)pg/ml,SAC组分别为(7.71±0.65)、(5.10±1.33)、(23.77±6.29)pg/ml,PAC组分别为(3.30±1.50)、(2.43±1.28)和(17.67±4.28)pg/ml,组间总体比较差异均有统计学意义( F=200.29、260.49、128.23,均 P<0.01);其中VKC组IL-4、IL-5、IL-13蛋白质量浓度明显高于SAC组和PAC组,SAC组IL-4、IL-5和IL-13蛋白质量浓度明显高于PAC组,差异均有统计学意义(均 P<0.01)。

结论IL-4、IL-5和IL-13参与多种类型过敏性结膜炎的发病,但在不同类型过敏性结膜炎患者中眼表的表达均有差异。

过敏性结膜炎/免疫;T淋巴细胞;眼部炎症/免疫;白细胞介素-4;白细胞介素-5;白细胞介素-13;人;上皮细胞/眼;泪液
ABSTRACT

BackgroundThe pathogenesis of allergic conjunctivitis has not been clearly established.Current researchers indicate that interleukin-4 (IL-4), IL-5 and IL-13 may play an important role in allergic conjunctivitis.But whether the roles of these inflammatory factors are same in different types of allergic conjunctivitis remains unclear.

ObjectiveThis study was to investigate the expressions of IL-4, IL-5 and IL-13 in ocular surface with different types of allergic conjunctivitis.

MethodsA prospective cohort study was designed.Eighty individuals were recruited in Shanxi Eye Hospital from April 2013 to September 2014, including 20 patients with vernal keratoconjunctivitis (VKC), 20 patients with seasonal allergic conjunctivitis (SAC), 20 patients with perennial allergic conjunctivitis (PAC) and 20 normal healthy subjects.Surficial tissues were binocularly scraped using disinfected scraper from upper eyelid conjunctiva, and 4 μl of tear fluid was obtained with capillary tube.The expressions of IL-4, IL-5 and IL-13 protein and mRNA in the conjunctival epithelial cells were detected by immunohistochemistry and real-time fluorescence quantitative PCR.The IL-4, IL-5 and IL-13 concentrations in tear fluid were assayed by Luminex method.This study complied with Declaration of Helsinki and the research protocol was approved by the Shanxi Eye Hospital Ethics Committee.Written informed consent was obtained from each subject prior to entering the cohort.

ResultsIL-4, IL-5 and IL-13 were positively expressed in cytoplasm of conjunctival epithelial cells in the VKC group, SAC group and PAC group, but the expressions of IL-4, IL-5 and IL-13 were absent in the normal control group.The relative expression levels of IL-4 mRNA were 4.11±1.24, 2.71±0.71 and 2.00±0.80; the relative expression levels of IL-5 mRNA were 4.02±0.43, 2.07±0.45 and 1.47±0.50; and the relative expression levels of IL-13 mRNA expression levels were 6.44±0.66, 4.35±1.26 and 2.39±0.86 in the VKC group, SAC group and PAC group, showing significant differences among the 4 groups ( F=51.32, 220.18, 162.49, all at P<0.01). The relative expression levels of IL-4, IL-5 and IL-13 mRNA were significantly higher in the VKC group than those in the SAC group and PAC group; and those in the SAC group were significantly elevated in comparison with the PAC group (all at P<0.05). No IL-4, IL-5 and IL-13 were detected in the tear fluid in the normal control group; while the concentrations of IL-4, IL-5 and IL-13 in the tear fluid were (14.06±3.50), (10.88±1.82) and (34.28±8.42) pg/ml in the VKC group, and (7.71±0.65), (5.10±1.33), (23.77±6.29) pg/ml in the SAC group as well as (3.30±1.50) pg/ml, (2.43±1.28) pg/ml and (17.67±4.28) pg/ml in the PAC group, showing significant differences among the 3 groups ( F=200.29, 260.49, 128.23, all at P<0.01). IL-4, IL-5 and IL-13 concentrations in the tear fluid were significantly higher in the VKC group than those in the SAC group and PAC group, and those in the SAC group were significantly raised in comparison with the PAC group (all at P<0.01).

ConclusionsIL-4, IL-5 and IL-13 participate in the pathogenesis of multiple allergic conjunctivitis, but their expressions in the ocular surficial tissue are discriminatory in different types of allergic conjunctivitis.

Conjunctivitis, allergic/immunology;T-lymphocytes;Inflammatory, ocular/immunology;Interleukin-4;Interleukin-5;Interleukin-13;Humans;Epithelium, eye;Tear
Zheng Xiaofen, Email: mocdef.3ab61dffxz
引用本文

姚娟,李冰,郑晓汾. 不同类型过敏性结膜炎患者眼表组织中白细胞介素-4、5和13的表达[J]. 中华实验眼科杂志,2015,33(10):924-929.

DOI:10.3760/cma.j.issn.2095-0160.2015.10.013

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过敏性结膜炎是局部组织和机体对抗原物质的非特异性免疫反应,由IgE或T淋巴细胞介导,其主要症状为眼痒和异物感。研究发现,过敏反应是由抗原激活的过度放大的免疫级联反应,导致辅助性T细胞1(T helper cell 1,Th1)/Th2平衡的破坏,主要是Th2细胞增多 [ 1 , 2 ]。抗原特异的Th2细胞分泌白细胞介素-4(interleukin-4,IL-4)、IL-5、IL-13等细胞因子 [ 3 ],促进IgE的合成,诱导嗜酸性粒细胞的增多及黏液分泌的增加,从而在过敏性结膜炎的发病过程中发挥作用 [ 4 ]。关于过敏性结膜炎的发病机制至今还有许多问题有待阐明,Th2细胞亚群在过敏性疾病中的作用是目前研究的热点之一。以往关于过敏性结膜炎的研究多聚焦于基础实验阶段,而对IL-4、IL-5、IL-13等重要信号传递分子在眼表的表达变化研究甚少。中国研究者曾对重度春季角结膜炎(vernal keratoconjunctivitis,VKC)患者的局部组织进行组织病理学研究,但缺乏对轻中度过敏性结膜炎患者的相关研究。本研究中检测临床上较常见的3种类型过敏性结膜炎患者结膜细胞中IL-4、IL-5和IL-13的表达情况,探讨Th2细胞亚群相关因子在过敏性角结膜炎发生和进展中的作用。
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郑晓汾,Email: mocdef.3ab61dffxz
B
山西省自然科学基金项目 (2013011059-6)
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