中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
Chinese Journal of Experimental Ophthalmology
2015年
10期
924-929
,共6页
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
過敏性結膜炎/免疫%T淋巴細胞%眼部炎癥/免疫%白細胞介素-4%白細胞介素-5%白細胞介素-13%人%上皮細胞/眼%淚液
과민성결막염/면역%T림파세포%안부염증/면역%백세포개소-4%백세포개소-5%백세포개소-13%인%상피세포/안%루액
Conjunctivitis,allergic/immunology%T-lymphocytes%Inflammatory,ocular/immunology%Interleukin-4%Interleukin-5%Interleukin-13%Humans%Epithelium,eye%Tear
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1μl毛细玻璃管于受试者下睑颞侧睑缘处吸取泪液4μl,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.
揹景 過敏性結膜炎的髮病機製尚未完全明確.研究錶明,白細胞介素-4(IL-4)、IL-5和IL-13在過敏性結膜炎中髮揮重要作用,但這些因子對不同類型的過敏性結膜炎患者的作用是否一緻尚有待證明.目的 探討IL-4、IL-5和IL-13在不同類型過敏性結膜炎患者眼錶組織中的錶達及其在不同類型過敏性結膜炎髮病中的作用.方法 採用前瞻性隊列研究設計.納入2013年4月至2014年9月在山西省眼科醫院就診的春季角結膜炎(VKC)患者20例40眼、季節性過敏性結膜炎(SAC)患者20例40眼和常年性過敏性結膜炎(PAC)患者20例40眼,併納入同期健康誌願者20人40眼作為正常對照.用消毒颳刀在上瞼結膜颳取結膜上皮細胞,分彆採用免疫組織化學法、實時熒光定量PCR法檢測結膜上皮細胞中IL-4、IL-5和IL-13蛋白及其mRNA的錶達;採用1μl毛細玻璃管于受試者下瞼顳側瞼緣處吸取淚液4μl,Luminex液相芯片測定受試者淚液中IL-4、IL-5和IL-13的錶達.結果 免疫組織化學法染色錶明,VKC組、SAC組和PAC組患者結膜上皮細胞的細胞質中均可見到IL-4、IL-5和IL-13的暘性錶達,而在正常對照組受試者結膜上皮細胞中未髮現IL-4、IL-5和IL-13的錶達.以正常對照組IL-4、IL-5和IL-13 mRNA的錶達量為對照,VKC組、SAC組和PAC組中IL-4 mRNA的相對錶達量分彆為4.11±1.24、2.71±0.71和2.00±0.80,IL-5 mRNA的相對錶達量分彆為4.02±0.43、2.07±0.45和1.47±0.50,IL-13 mRNA的相對錶達量分彆為6.44±0.66、4.35±1.26和2.39±0.86,4箇組間IL-4、IL-5和IL-13 mRNA相對錶達量的總體比較差異均有統計學意義(F=51.32、220.18、162.49,均P＜0.01);其中VKC組IL-4、IL-5和IL-13 mRNA的相對錶達量明顯高于SAC組和PAC組,SAC組IL-4、IL-5、IL-13 mRNA的相對錶達量高于PAC組,差異均有統計學意義(均P＜0.05).正常對照組受試者淚液中未檢測到IL-4、IL-5和IL-13錶達,VKC組淚液中IL-4、IL-5和IL-13蛋白質量濃度分彆為(14.06±3.50)、(10.88±1.82)和(34.28±8.42) pg/ml,SAC組分彆為(7.71±0.65)、(5.10±1.33)、(23.77±6.29) pg/ml,PAC組分彆為(3.30±1.50)、(2.43±1.28)和(17.67±4.28) pg/ml,組間總體比較差異均有統計學意義(F=200.29、260.49、128.23,均P＜0.01);其中VKC組IL-4、IL-5、IL-13蛋白質量濃度明顯高于SAC組和PAC組,SAC組IL-4、IL-5和IL-13蛋白質量濃度明顯高于PAC組,差異均有統計學意義(均P＜0.01).結論 IL-4、IL-5和IL-13參與多種類型過敏性結膜炎的髮病,但在不同類型過敏性結膜炎患者中眼錶的錶達均有差異.
배경 과민성결막염적발병궤제상미완전명학.연구표명,백세포개소-4(IL-4)、IL-5화IL-13재과민성결막염중발휘중요작용,단저사인자대불동류형적과민성결막염환자적작용시부일치상유대증명.목적 탐토IL-4、IL-5화IL-13재불동류형과민성결막염환자안표조직중적표체급기재불동류형과민성결막염발병중적작용.방법 채용전첨성대렬연구설계.납입2013년4월지2014년9월재산서성안과의원취진적춘계각결막염(VKC)환자20례40안、계절성과민성결막염(SAC)환자20례40안화상년성과민성결막염(PAC)환자20례40안,병납입동기건강지원자20인40안작위정상대조.용소독괄도재상검결막괄취결막상피세포,분별채용면역조직화학법、실시형광정량PCR법검측결막상피세포중IL-4、IL-5화IL-13단백급기mRNA적표체;채용1μl모세파리관우수시자하검섭측검연처흡취루액4μl,Luminex액상심편측정수시자루액중IL-4、IL-5화IL-13적표체.결과 면역조직화학법염색표명,VKC조、SAC조화PAC조환자결막상피세포적세포질중균가견도IL-4、IL-5화IL-13적양성표체,이재정상대조조수시자결막상피세포중미발현IL-4、IL-5화IL-13적표체.이정상대조조IL-4、IL-5화IL-13 mRNA적표체량위대조,VKC조、SAC조화PAC조중IL-4 mRNA적상대표체량분별위4.11±1.24、2.71±0.71화2.00±0.80,IL-5 mRNA적상대표체량분별위4.02±0.43、2.07±0.45화1.47±0.50,IL-13 mRNA적상대표체량분별위6.44±0.66、4.35±1.26화2.39±0.86,4개조간IL-4、IL-5화IL-13 mRNA상대표체량적총체비교차이균유통계학의의(F=51.32、220.18、162.49,균P＜0.01);기중VKC조IL-4、IL-5화IL-13 mRNA적상대표체량명현고우SAC조화PAC조,SAC조IL-4、IL-5、IL-13 mRNA적상대표체량고우PAC조,차이균유통계학의의(균P＜0.05).정상대조조수시자루액중미검측도IL-4、IL-5화IL-13표체,VKC조루액중IL-4、IL-5화IL-13단백질량농도분별위(14.06±3.50)、(10.88±1.82)화(34.28±8.42) pg/ml,SAC조분별위(7.71±0.65)、(5.10±1.33)、(23.77±6.29) pg/ml,PAC조분별위(3.30±1.50)、(2.43±1.28)화(17.67±4.28) pg/ml,조간총체비교차이균유통계학의의(F=200.29、260.49、128.23,균P＜0.01);기중VKC조IL-4、IL-5、IL-13단백질량농도명현고우SAC조화PAC조,SAC조IL-4、IL-5화IL-13단백질량농도명현고우PAC조,차이균유통계학의의(균P＜0.01).결론 IL-4、IL-5화IL-13삼여다충류형과민성결막염적발병,단재불동류형과민성결막염환자중안표적표체균유차이.
Background The 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.Objective This study was to investigate the expressions of IL-4, IL-5 and IL-13 in ocular surface with different types of allergic conjunctivitis.Methods A 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.Results IL-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).Conclusions IL-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.
